                         SEQUENCE LISTING

<110>  Targetgene Biotechnologies Ltd
 
<120>  PAM-REDUCED AND PAM-ABOLISHED CAS DERIVATIVES COMPOSITIONS AND 
       USES THEREOF IN GENETIC MODULATION

<130>  2754312

<150>  US 62/926,764
<151>  2019-10-28

<150>  US 63/068,060
<151>  2020-08-20

<160>  506   

<170>  PatentIn version 3.5

<210>  1
<211>  8648
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Construct 10916 dScCas9-FokI EMX1sgRNA27T1-NNG

<400>  1
ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc       60

attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga      120

gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt      180

gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt      240

gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg      300

acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca      360

aggccgcata tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg      420

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt      480

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca      540

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc      600

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta      660

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac      720

catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg      780

atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg      840

ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt      900

acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta      960

ccggtatggc ccctaagaag aagagaaaag tcggcggctc tggcagccag ctggtcaagt     1020

ctgagctgga agagaagaaa agcgagctgc ggcacaagct gaaatacgtg ccccacgagt     1080

acatcgagct gatcgagatc gccagaaaca gcacccagga cagaatcctg gaaatgaagg     1140

tcatggaatt ctttatgaag gtgtacggct accggggcaa gcacctcgga ggatctagaa     1200

agcctgacgg cgccatctat accgtgggca gccctatcga ttacggcgtg atcgtggata     1260

ccaaggccta cagcggcggc tacaatctgc ctattggaca ggccgacgag atgcagagat     1320

acgtggaaga aaaccagaca cggaacaagc acatcaaccc caacgagtgg tggaaagtgt     1380

accccagcag cgtgaccgag ttcaagttcc tgttcgtgtc cggccacttc aagggcaact     1440

acaaggccca gctgacccgg ctgaaccaca tcacaaattg caacggcgct gtgctgtctg     1500

tggaagaact gctgatcggc ggcgagatga tcaaggccgg aacactgaca ctggaagaag     1560

tgcggcggaa gttcaacaac ggcgagatca acttcagcgg cagcgagaca cctggcacat     1620

ctgaatctgc cacacctgag agcatggaaa agaagtacag catcggcctg gccatcggca     1680

caaattctgt tggctgggcc gtgatcaccg acgactacaa ggtgcccagc aagaaattca     1740

aggtgctggg caacaccaac cggaagtcca tcaagaaaaa cctgatgggc gccctgctgt     1800

tcgattctgg cgaaacagcc gaggccacca gactgaagag aaccgccaga agaagataca     1860

cccggcggaa gaaccggatc agatacctgc aagagatctt cgccaacgag atggccaagc     1920

tggacgacag cttctttcag agactggaag agtccttcct ggtggaagag gacaagaaga     1980

acgagcggca ccccatcttc ggcaatctgg ctgatgaggt ggcctaccac agaaactacc     2040

ccaccatcta ccacctgagg aagaagctgg ccgactctcc tgagaaggcc gacctgagac     2100

tgatctatct ggccctggct cacatcatca agttccgggg ccacttcctg atcgagggca     2160

agctgaatgc cgagaacagc gacgtggcca aactgttcta ccagctgatc cagacctaca     2220

accagctgtt cgaggaaagc cctctggacg agattgaggt ggacgccaag ggaatcctga     2280

gcgccagact gagcaagagc aagcggctgg aaaagctgat cgccgtgttt cccaacgaga     2340

agaagaatgg cctgttcggc aacatcattg ccctggcact gggcctgaca cctaacttca     2400

agtccaactt cgacctgacc gaggacgcca aactgcagct gagcaaggac acctacgacg     2460

acgatctgga tgagctgctg ggccagatcg gagatcagta cgccgatctg ttcagcgccg     2520

ccaagaatct gtctgacgcc atcctgctga gcgacatcct gagatccaac agcgaagtga     2580

caaaggcccc tctgagcgcc tccatggtca agagatatga cgagcaccac caggacctgg     2640

ctctgctgaa aacccttgtg cggcagcagt tccccgagaa gtatgccgag atcttcaagg     2700

acgacaccaa gaacggctac gccggctatg ttggcatcgg catcaagcac agaaagcgga     2760

ccaccaagct ggccacacaa gaggaattct acaagttcat caagcccatc ctcgagaaga     2820

tggacggcgc cgaagaactc ctggctaagc tgaacaggga cgacctgctg cggaagcaga     2880

gaaccttcga caatggcagc atccctcacc agatccacct gaaagagctg cacgccattc     2940

tgcggcggca agaagagttc tacccattcc tgaaagaaaa ccgggaaaag attgagaaga     3000

tcctgacctt caggatcccc tactacgtgg gaccactggc cagaggcaat agcagattcg     3060

cttggctgac cagaaagagc gaggaagcca tcacaccctg gaacttcgag gaagtggtgg     3120

ataagggcgc cagcgctcag tccttcatcg agcggatgac caatttcgac gagcagctgc     3180

ccaacaagaa ggtgctgcct aagcacagcc tgctgtacga gtacttcacc gtgtacaatg     3240

agctgaccaa agtcaaatac gtgacagagc ggatgcggaa gcctgagttt ctgagcggcg     3300

agcagaaaaa ggccatcgtg gatctgctgt tcaagaccaa cagaaaagtg accgtgaagc     3360

agctgaaaga ggactacttc aagaaaatcg agtgcttcga cagcgtcgag atcatcggcg     3420

tggaagatag attcaacgcc agcctgggca cataccacga tctgctgaag attatcaagg     3480

acaaggactt cctggacaac gaggaaaacg aggacatcct ggaagatatc gtgctgaccc     3540

tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacatac gcccacctgt     3600

tcgacgacaa agtgatgaag caactgaagc ggcggcacta cacaggctgg ggcagactgt     3660

ccagaaagat gattaacggc atccgggaca agcagagcgg caagaccatc ctggactttc     3720

tgaagtccga cggcttcagc aaccggaact tcatgcagct gattcacgac gacagcctca     3780

ccttcaaaga ggaaatcgag aaggcccagg tttccggcca gggcgattct ctgcacgagc     3840

agattgctga tctggccgga tctcccgcca ttaagaaggg catcctgcag acagtgaaga     3900

tcgtggacga actcgtgaaa gtgatgggcc acaagcctga gaacatcgtg atcgaaatgg     3960

ccagagagaa ccagaccacc accaagggcc tgcagcagtc cagagagaga aagaagcgga     4020

tcgaagaggg catcaaagag ctggaaagcc agatcctcaa agagaacccc gtcgagaaca     4080

cccagctgca gaacgagaag ctgtacctgt actacctgca gaatggacgg gatatgtacg     4140

tggaccaaga actggacatc aaccggctgt ccgactacga tgtggatgcc atcgtgcccc     4200

agagcttcat caaggatgac agcatcgaca acaaggtcct gaccagatcc gtggaaaaca     4260

gaggcaagag cgacaacgtg ccctccgaag aggtggtcaa gaagatgaag aattactggc     4320

gacagctgct gaacgccaag ctgatcaccc agagaaagtt cgacaacctg accaaggccg     4380

agagaggcgg actgtctgaa gctgataagg ccggcttcat taagcggcag ctggtggaaa     4440

cccggcagat cacaaagcac gtggcccgga tcctggacag cagaatgaac accaagcggg     4500

acaagaacga caagcctatc cgggaagtga aagtcatcac cctgaagtcc aagctggtgt     4560

ccgatttccg gaaggatttc cagctgtaca aagtgcgcga catcaacaac taccatcacg     4620

cccacgacgc ctacctgaat gctgttgtgg gaacagccct gatcaagaag taccctaagc     4680

tggaatccga gttcgtgtac ggcgactaca aagtgtacga cgtgcggaag atgatcgcca     4740

agagcgagca agagattggc aaggccaccg ccaagcggtt cttctactcc aacatcatga     4800

actttttcaa gaccgaagtg aagctcgcca acggcgaaat ccggaagagg cccctgattg     4860

agacaaatgg cgagacaggc gaggtcgtgt ggaacaaaga gaaggacttc gccaccgtgc     4920

ggaaagtgct ggccatgcct caagtgaata tcgtgaaaaa gaccgaggtg cagaccggcg     4980

gcttctccaa agagagcatc ctgtctaaga gagagagcgc caagctcatc cctcggaaga     5040

aaggctggga tacacggaag tacggcggat ttggaagccc taccgtggcc tactctattc     5100

tggtggtggc caaggtggaa aagggcaaag ccaagaaact gaagtctgtg aaggtgctcg     5160

tggggatcac catcatggaa aaaggcagct acgagaagga ccctatcggc tttctggaag     5220

ccaagggcta caaggacatc aaaaaagagc tgatctttaa gctccccaag tacagcctgt     5280

tcgaactgga aaacggcaga cgcagaatgc tggcctctgc cacagaactg cagaaagcca     5340

atgaactggt gctgccacaa cacctcgtgc ggctgctgta ctacacccag aatatcagcg     5400

ccaccaccgg cagcaacaac ctgggatata tcgagcagca ccgggaagag ttcaaagaga     5460

ttttcgagaa aatcatcgac ttcagcgaga agtacatcct gaagaacaaa gtgaacagca     5520

acctcaagag cagcttcgat gagcagttcg ccgtgtccga ctctatcctg ctgtctaaca     5580

gcttcgtgtc cctgctgaag tacacctcct ttggcgcctc tggcggcttc acctttctgg     5640

atctggacgt gaaacagggg agactgagat accagaccgt gactgaggtg ctggacgcca     5700

ctctgatcta ccagagcatc accggcctgt acgagacaag aaccgatctc tctcagctcg     5760

gaggcgacta aaagcttctg tgccttctag ttgccagcca tctgttgttt gcccctcccc     5820

cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga     5880

aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga     5940

cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat     6000

ggacaaaccc acaactacgc gtaaaaaaaa agcaccgact cggtgccact ttttcaagtt     6060

gataacggac tagccttatt ttaacttgct atttctagct ctaaaactcc tctagaaact     6120

cgtagagggt gtttcgtggt ctcatacaga acttataaga ttcccaaatc caaagacatt     6180

tcacgtttat ggtgatttcc cagaacacat agcgacatgc aaatattgca gggcgccact     6240

cccctgtccc tcacagccat cttcctgcca gggcgcacgc gcgctgggtg ttcccgccta     6300

gtgacactgg gcccgcgatt ccttggagcg ggttgatgac gtcagcgttc aaagagggcc     6360

tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg     6420

gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat     6480

aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta     6540

ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca     6600

cctgcactcc tccaccttgg ctgttttaga gctagaaata gcaagttaaa ataaggctag     6660

tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttacgcg tacttgcggc     6720

cgcatattcc taggatattc tgggcctcat gggccttcct ttcactgccc gctttccagt     6780

cgggaaacct gtcgtgccag ctgcattaac atggtcatag ctgtttcctt gcgtattggg     6840

cgctctccgc ttcctcgctc actgactcgc tgcgctcggt cgttcgggta aagcctgggg     6900

tgcctaatga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc     6960

gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag     7020

gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt     7080

gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg     7140

aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg     7200

ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg     7260

taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac     7320

tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg     7380

gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt     7440

taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg     7500

tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc     7560

tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt     7620

ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt     7680

taaatcaatc taaagtatat atgagtaaac ttggtctgac agttattaga aaaattcatc     7740

cagcagacga taaaacgcaa tacgctggct atccggtgcc gcaatgccat acagcaccag     7800

aaaacgatcc gcccattcgc cgcccagttc ttccgcaata tcacgggtgg ccagcgcaat     7860

atcctgataa cgatccgcca cgcccagacg gccgcaatca ataaagccgc taaaacggcc     7920

attttccacc ataatgttcg gcaggcacgc atcaccatgg gtcaccacca gatcttcgcc     7980

atccggcatg ctcgctttca gacgcgcaaa cagctctgcc ggtgccaggc cctgatgttc     8040

ttcatccaga tcatcctgat ccaccaggcc cgcttccata cgggtacgcg cacgttcaat     8100

acgatgtttc gcctgatgat caaacggaca ggtcgccggg tccagggtat gcagacgacg     8160

catggcatcc gccataatgc tcactttttc tgccggcgcc agatggctag acagcagatc     8220

ctgacccggc acttcgccca gcagcagcca atcacggccc gcttcggtca ccacatccag     8280

caccgccgca cacggaacac cggtggtggc cagccagctc agacgcgccg cttcatcctg     8340

cagctcgttc agcgcaccgc tcagatcggt tttcacaaac agcaccggac gaccctgcgc     8400

gctcagacga aacaccgccg catcagagca gccaatggtc tgctgcgccc aatcatagcc     8460

aaacagacgt tccacccacg ctgccgggct acccgcatgc aggccatcct gttcaatcat     8520

actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata     8580

catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa     8640

agtgccac                                                              8648


<210>  2
<211>  1601
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI

<400>  2

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
                805                 810                 815     


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
            820                 825                 830         


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
        835                 840                 845             


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
    850                 855                 860                 


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
865                 870                 875                 880 


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
                885                 890                 895     


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
            900                 905                 910         


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
        915                 920                 925             


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
    930                 935                 940                 


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
945                 950                 955                 960 


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
                965                 970                 975     


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
            980                 985                 990         


Asn Ile Val Ile Glu Met Ala Arg  Glu Asn Gln Thr Thr  Thr Lys Gly 
        995                 1000                 1005             


Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg Ile Glu  Glu Gly Ile 
    1010                 1015                 1020             


Lys Glu  Leu Glu Ser Gln Ile  Leu Lys Glu Asn Pro  Val Glu Asn 
    1025                 1030                 1035             


Thr Gln  Leu Gln Asn Glu Lys  Leu Tyr Leu Tyr Tyr  Leu Gln Asn 
    1040                 1045                 1050             


Gly Arg  Asp Met Tyr Val Asp  Gln Glu Leu Asp Ile  Asn Arg Leu 
    1055                 1060                 1065             


Ser Asp  Tyr Asp Val Asp Ala  Ile Val Pro Gln Ser  Phe Ile Lys 
    1070                 1075                 1080             


Asp Asp  Ser Ile Asp Asn Lys  Val Leu Thr Arg Ser  Val Glu Asn 
    1085                 1090                 1095             


Arg Gly  Lys Ser Asp Asn Val  Pro Ser Glu Glu Val  Val Lys Lys 
    1100                 1105                 1110             


Met Lys  Asn Tyr Trp Arg Gln  Leu Leu Asn Ala Lys  Leu Ile Thr 
    1115                 1120                 1125             


Gln Arg  Lys Phe Asp Asn Leu  Thr Lys Ala Glu Arg  Gly Gly Leu 
    1130                 1135                 1140             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1145                 1150                 1155             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1160                 1165                 1170             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1175                 1180                 1185             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1190                 1195                 1200             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1205                 1210                 1215             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1220                 1225                 1230             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1235                 1240                 1245             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1250                 1255                 1260             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1265                 1270                 1275             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1280                 1285                 1290             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1295                 1300                 1305             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1310                 1315                 1320             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1325                 1330                 1335             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1340                 1345                 1350             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1355                 1360                 1365             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1370                 1375                 1380             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1385                 1390                 1395             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1400                 1405                 1410             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1415                 1420                 1425             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1430                 1435                 1440             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1445                 1450                 1455             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1460                 1465                 1470             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1475                 1480                 1485             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1490                 1495                 1500             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1505                 1510                 1515             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1520                 1525                 1530             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1535                 1540                 1545             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1550                 1555                 1560             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1565                 1570                 1575             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1580                 1585                 1590             


Asp Leu  Ser Gln Leu Gly Gly  Asp 
    1595                 1600     


<210>  3
<211>  697
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX1sgRNA27T1-NNG

<400>  3
tacgcgtaaa aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc       60

ttattttaac ttgctatttc tagctctaaa actcctctag aaactcgtag agggtgtttc      120

gtggtctcat acagaactta taagattccc aaatccaaag acatttcacg tttatggtga      180

tttcccagaa cacatagcga catgcaaata ttgcagggcg ccactcccct gtccctcaca      240

gccatcttcc tgccagggcg cacgcgcgct gggtgttccc gcctagtgac actgggcccg      300

cgattccttg gagcgggttg atgacgtcag cgttcaaaga gggcctattt cccatgattc      360

cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg      420

taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt      480

ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt      540

atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacacctgc actcctccac      600

cttggctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg      660

aaaaagtggc accgagtcgg tgcttttttt acgcgta                               697


<210>  4
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX1SGRNA15T1-NNG

<400>  4
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa ctagaaactc gtagagtccc aggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccagag cctgcactcc      600

tccaccgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  5
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3157

<400>  5
attagttcat agcccatata tggagttc                                          28


<210>  6
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3158

<400>  6
tacgcgtagt tgtgggtttg tc                                                22


<210>  7
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3206

<400>  7
tccagagaga gaaagaagcg cggcggactg tctgaagctg at                          42


<210>  8
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3207

<400>  8
atcagcttca gacagtccgc cgcgcttctt tctctctctg ga                          42


<210>  9
<211>  8282
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Plasmid 11248 dScCas9-FokI dHNH-F-NNG EMX1 T1 sgRNA 27

<400>  9
ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc       60

attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga      120

gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt      180

gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt      240

gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg      300

acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca      360

aggccgcata tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg      420

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt      480

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca      540

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc      600

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta      660

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac      720

catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg      780

atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg      840

ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt      900

acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta      960

ccggtatggc ccctaagaag aagagaaaag tcggcggctc tggcagccag ctggtcaagt     1020

ctgagctgga agagaagaaa agcgagctgc ggcacaagct gaaatacgtg ccccacgagt     1080

acatcgagct gatcgagatc gccagaaaca gcacccagga cagaatcctg gaaatgaagg     1140

tcatggaatt ctttatgaag gtgtacggct accggggcaa gcacctcgga ggatctagaa     1200

agcctgacgg cgccatctat accgtgggca gccctatcga ttacggcgtg atcgtggata     1260

ccaaggccta cagcggcggc tacaatctgc ctattggaca ggccgacgag atgcagagat     1320

acgtggaaga aaaccagaca cggaacaagc acatcaaccc caacgagtgg tggaaagtgt     1380

accccagcag cgtgaccgag ttcaagttcc tgttcgtgtc cggccacttc aagggcaact     1440

acaaggccca gctgacccgg ctgaaccaca tcacaaattg caacggcgct gtgctgtctg     1500

tggaagaact gctgatcggc ggcgagatga tcaaggccgg aacactgaca ctggaagaag     1560

tgcggcggaa gttcaacaac ggcgagatca acttcagcgg cagcgagaca cctggcacat     1620

ctgaatctgc cacacctgag agcatggaaa agaagtacag catcggcctg gccatcggca     1680

caaattctgt tggctgggcc gtgatcaccg acgactacaa ggtgcccagc aagaaattca     1740

aggtgctggg caacaccaac cggaagtcca tcaagaaaaa cctgatgggc gccctgctgt     1800

tcgattctgg cgaaacagcc gaggccacca gactgaagag aaccgccaga agaagataca     1860

cccggcggaa gaaccggatc agatacctgc aagagatctt cgccaacgag atggccaagc     1920

tggacgacag cttctttcag agactggaag agtccttcct ggtggaagag gacaagaaga     1980

acgagcggca ccccatcttc ggcaatctgg ctgatgaggt ggcctaccac agaaactacc     2040

ccaccatcta ccacctgagg aagaagctgg ccgactctcc tgagaaggcc gacctgagac     2100

tgatctatct ggccctggct cacatcatca agttccgggg ccacttcctg atcgagggca     2160

agctgaatgc cgagaacagc gacgtggcca aactgttcta ccagctgatc cagacctaca     2220

accagctgtt cgaggaaagc cctctggacg agattgaggt ggacgccaag ggaatcctga     2280

gcgccagact gagcaagagc aagcggctgg aaaagctgat cgccgtgttt cccaacgaga     2340

agaagaatgg cctgttcggc aacatcattg ccctggcact gggcctgaca cctaacttca     2400

agtccaactt cgacctgacc gaggacgcca aactgcagct gagcaaggac acctacgacg     2460

acgatctgga tgagctgctg ggccagatcg gagatcagta cgccgatctg ttcagcgccg     2520

ccaagaatct gtctgacgcc atcctgctga gcgacatcct gagatccaac agcgaagtga     2580

caaaggcccc tctgagcgcc tccatggtca agagatatga cgagcaccac caggacctgg     2640

ctctgctgaa aacccttgtg cggcagcagt tccccgagaa gtatgccgag atcttcaagg     2700

acgacaccaa gaacggctac gccggctatg ttggcatcgg catcaagcac agaaagcgga     2760

ccaccaagct ggccacacaa gaggaattct acaagttcat caagcccatc ctcgagaaga     2820

tggacggcgc cgaagaactc ctggctaagc tgaacaggga cgacctgctg cggaagcaga     2880

gaaccttcga caatggcagc atccctcacc agatccacct gaaagagctg cacgccattc     2940

tgcggcggca agaagagttc tacccattcc tgaaagaaaa ccgggaaaag attgagaaga     3000

tcctgacctt caggatcccc tactacgtgg gaccactggc cagaggcaat agcagattcg     3060

cttggctgac cagaaagagc gaggaagcca tcacaccctg gaacttcgag gaagtggtgg     3120

ataagggcgc cagcgctcag tccttcatcg agcggatgac caatttcgac gagcagctgc     3180

ccaacaagaa ggtgctgcct aagcacagcc tgctgtacga gtacttcacc gtgtacaatg     3240

agctgaccaa agtcaaatac gtgacagagc ggatgcggaa gcctgagttt ctgagcggcg     3300

agcagaaaaa ggccatcgtg gatctgctgt tcaagaccaa cagaaaagtg accgtgaagc     3360

agctgaaaga ggactacttc aagaaaatcg agtgcttcga cagcgtcgag atcatcggcg     3420

tggaagatag attcaacgcc agcctgggca cataccacga tctgctgaag attatcaagg     3480

acaaggactt cctggacaac gaggaaaacg aggacatcct ggaagatatc gtgctgaccc     3540

tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacatac gcccacctgt     3600

tcgacgacaa agtgatgaag caactgaagc ggcggcacta cacaggctgg ggcagactgt     3660

ccagaaagat gattaacggc atccgggaca agcagagcgg caagaccatc ctggactttc     3720

tgaagtccga cggcttcagc aaccggaact tcatgcagct gattcacgac gacagcctca     3780

ccttcaaaga ggaaatcgag aaggcccagg tttccggcca gggcgattct ctgcacgagc     3840

agattgctga tctggccgga tctcccgcca ttaagaaggg catcctgcag acagtgaaga     3900

tcgtggacga actcgtgaaa gtgatgggcc acaagcctga gaacatcgtg atcgaaatgg     3960

ccagagagaa ccagaccacc accaagggcc tgcagcagtc cagagagaga aagaagcgcg     4020

gcggactgtc tgaagctgat aaggccggct tcattaagcg gcagctggtg gaaacccggc     4080

agatcacaaa gcacgtggcc cggatcctgg acagcagaat gaacaccaag cgggacaaga     4140

acgacaagcc tatccgggaa gtgaaagtca tcaccctgaa gtccaagctg gtgtccgatt     4200

tccggaagga tttccagctg tacaaagtgc gcgacatcaa caactaccat cacgcccacg     4260

acgcctacct gaatgctgtt gtgggaacag ccctgatcaa gaagtaccct aagctggaat     4320

ccgagttcgt gtacggcgac tacaaagtgt acgacgtgcg gaagatgatc gccaagagcg     4380

agcaagagat tggcaaggcc accgccaagc ggttcttcta ctccaacatc atgaactttt     4440

tcaagaccga agtgaagctc gccaacggcg aaatccggaa gaggcccctg attgagacaa     4500

atggcgagac aggcgaggtc gtgtggaaca aagagaagga cttcgccacc gtgcggaaag     4560

tgctggccat gcctcaagtg aatatcgtga aaaagaccga ggtgcagacc ggcggcttct     4620

ccaaagagag catcctgtct aagagagaga gcgccaagct catccctcgg aagaaaggct     4680

gggatacacg gaagtacggc ggatttggaa gccctaccgt ggcctactct attctggtgg     4740

tggccaaggt ggaaaagggc aaagccaaga aactgaagtc tgtgaaggtg ctcgtgggga     4800

tcaccatcat ggaaaaaggc agctacgaga aggaccctat cggctttctg gaagccaagg     4860

gctacaagga catcaaaaaa gagctgatct ttaagctccc caagtacagc ctgttcgaac     4920

tggaaaacgg cagacgcaga atgctggcct ctgccacaga actgcagaaa gccaatgaac     4980

tggtgctgcc acaacacctc gtgcggctgc tgtactacac ccagaatatc agcgccacca     5040

ccggcagcaa caacctggga tatatcgagc agcaccggga agagttcaaa gagattttcg     5100

agaaaatcat cgacttcagc gagaagtaca tcctgaagaa caaagtgaac agcaacctca     5160

agagcagctt cgatgagcag ttcgccgtgt ccgactctat cctgctgtct aacagcttcg     5220

tgtccctgct gaagtacacc tcctttggcg cctctggcgg cttcaccttt ctggatctgg     5280

acgtgaaaca ggggagactg agataccaga ccgtgactga ggtgctggac gccactctga     5340

tctaccagag catcaccggc ctgtacgaga caagaaccga tctctctcag ctcggaggcg     5400

actaaaagct tctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc     5460

ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc     5520

atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa     5580

gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggacaa     5640

acccacaact acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac     5700

ggactagcct tattttaact tgctatttct agctctaaaa ctcctctaga aactcgtaga     5760

gggtgtttcg tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt     5820

ttatggtgat ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg     5880

tccctcacag ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca     5940

ctgggcccgc gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc     6000

ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta     6060

atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc     6120

ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa     6180

cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacacctgca     6240

ctcctccacc ttggctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt     6300

atcaacttga aaaagtggca ccgagtcggt gcttttttta cgcgtacttg cggccgcata     6360

ttcctaggat attctgggcc tcatgggcct tcctttcact gcccgctttc cagtcgggaa     6420

acctgtcgtg ccagctgcat taacatggtc atagctgttt ccttgcgtat tgggcgctct     6480

ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg ggtaaagcct ggggtgccta     6540

atgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt     6600

ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg     6660

aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc     6720

tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt     6780

ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa     6840

gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta     6900

tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa     6960

caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa     7020

ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt     7080

cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt     7140

ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat     7200

cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat     7260

gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc     7320

aatctaaagt atatatgagt aaacttggtc tgacagttat tagaaaaatt catccagcag     7380

acgataaaac gcaatacgct ggctatccgg tgccgcaatg ccatacagca ccagaaaacg     7440

atccgcccat tcgccgccca gttcttccgc aatatcacgg gtggccagcg caatatcctg     7500

ataacgatcc gccacgccca gacggccgca atcaataaag ccgctaaaac ggccattttc     7560

caccataatg ttcggcaggc acgcatcacc atgggtcacc accagatctt cgccatccgg     7620

catgctcgct ttcagacgcg caaacagctc tgccggtgcc aggccctgat gttcttcatc     7680

cagatcatcc tgatccacca ggcccgcttc catacgggta cgcgcacgtt caatacgatg     7740

tttcgcctga tgatcaaacg gacaggtcgc cgggtccagg gtatgcagac gacgcatggc     7800

atccgccata atgctcactt tttctgccgg cgccagatgg ctagacagca gatcctgacc     7860

cggcacttcg cccagcagca gccaatcacg gcccgcttcg gtcaccacat ccagcaccgc     7920

cgcacacgga acaccggtgg tggccagcca gctcagacgc gccgcttcat cctgcagctc     7980

gttcagcgca ccgctcagat cggttttcac aaacagcacc ggacgaccct gcgcgctcag     8040

acgaaacacc gccgcatcag agcagccaat ggtctgctgc gcccaatcat agccaaacag     8100

acgttccacc cacgctgccg ggctacccgc atgcaggcca tcctgttcaa tcatactctt     8160

cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt     8220

tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc     8280

ac                                                                    8282


<210>  10
<211>  1482
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI dHNH Protein sequence

<400>  10

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
                805                 810                 815     


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
            820                 825                 830         


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
        835                 840                 845             


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
    850                 855                 860                 


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
865                 870                 875                 880 


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
                885                 890                 895     


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
            900                 905                 910         


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
        915                 920                 925             


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
    930                 935                 940                 


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
945                 950                 955                 960 


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
                965                 970                 975     


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
            980                 985                 990         


Asn Ile Val Ile Glu Met Ala Arg  Glu Asn Gln Thr Thr  Thr Lys Gly 
        995                 1000                 1005             


Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg Gly Gly  Ser Gly Gly 
    1010                 1015                 1020             


Leu Ser  Glu Ala Asp Lys Ala  Gly Phe Ile Lys Arg  Gln Leu Val 
    1025                 1030                 1035             


Glu Thr  Arg Gln Ile Thr Lys  His Val Ala Arg Ile  Leu Asp Ser 
    1040                 1045                 1050             


Arg Met  Asn Thr Lys Arg Asp  Lys Asn Asp Lys Pro  Ile Arg Glu 
    1055                 1060                 1065             


Val Lys  Val Ile Thr Leu Lys  Ser Lys Leu Val Ser  Asp Phe Arg 
    1070                 1075                 1080             


Lys Asp  Phe Gln Leu Tyr Lys  Val Arg Asp Ile Asn  Asn Tyr His 
    1085                 1090                 1095             


His Ala  His Asp Ala Tyr Leu  Asn Ala Val Val Gly  Thr Ala Leu 
    1100                 1105                 1110             


Ile Lys  Lys Tyr Pro Lys Leu  Glu Ser Glu Phe Val  Tyr Gly Asp 
    1115                 1120                 1125             


Tyr Lys  Val Tyr Asp Val Arg  Lys Met Ile Ala Lys  Ser Glu Gln 
    1130                 1135                 1140             


Glu Ile  Gly Lys Ala Thr Ala  Lys Arg Phe Phe Tyr  Ser Asn Ile 
    1145                 1150                 1155             


Met Asn  Phe Phe Lys Thr Glu  Val Lys Leu Ala Asn  Gly Glu Ile 
    1160                 1165                 1170             


Arg Lys  Arg Pro Leu Ile Glu  Thr Asn Gly Glu Thr  Gly Glu Val 
    1175                 1180                 1185             


Val Trp  Asn Lys Glu Lys Asp  Phe Ala Thr Val Arg  Lys Val Leu 
    1190                 1195                 1200             


Ala Met  Pro Gln Val Asn Ile  Val Lys Lys Thr Glu  Val Gln Thr 
    1205                 1210                 1215             


Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser Lys Arg  Glu Ser Ala 
    1220                 1225                 1230             


Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp Thr Arg  Lys Tyr Gly 
    1235                 1240                 1245             


Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser Ile Leu  Val Val Ala 
    1250                 1255                 1260             


Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu Lys Ser  Val Lys Val 
    1265                 1270                 1275             


Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly Ser Tyr  Glu Lys Asp 
    1280                 1285                 1290             


Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr Lys Asp  Ile Lys Lys 
    1295                 1300                 1305             


Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser Leu Phe  Glu Leu Glu 
    1310                 1315                 1320             


Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala Thr Glu  Leu Gln Lys 
    1325                 1330                 1335             


Ala Asn  Glu Leu Val Leu Pro  Gln His Leu Val Arg  Leu Leu Tyr 
    1340                 1345                 1350             


Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly Ser Asn  Asn Leu Gly 
    1355                 1360                 1365             


Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys Glu Ile  Phe Glu Lys 
    1370                 1375                 1380             


Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu Lys Asn  Lys Val Asn 
    1385                 1390                 1395             


Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln Phe Ala  Val Ser Asp 
    1400                 1405                 1410             


Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser Leu Leu  Lys Tyr Thr 
    1415                 1420                 1425             


Ser Phe  Gly Ala Ser Gly Gly  Phe Thr Phe Leu Asp  Leu Asp Val 
    1430                 1435                 1440             


Lys Gln  Gly Arg Leu Arg Tyr  Gln Thr Val Thr Glu  Val Leu Asp 
    1445                 1450                 1455             


Ala Thr  Leu Ile Tyr Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1460                 1465                 1470             


Thr Asp  Leu Ser Gln Leu Gly  Gly Asp 
    1475                 1480         


<210>  11
<211>  697
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX1sgRNA27T1-NNG

<400>  11
tacgcgtaaa aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc       60

ttattttaac ttgctatttc tagctctaaa actcctctag aaactcgtag agggtgtttc      120

gtggtctcat acagaactta taagattccc aaatccaaag acatttcacg tttatggtga      180

tttcccagaa cacatagcga catgcaaata ttgcagggcg ccactcccct gtccctcaca      240

gccatcttcc tgccagggcg cacgcgcgct gggtgttccc gcctagtgac actgggcccg      300

cgattccttg gagcgggttg atgacgtcag cgttcaaaga gggcctattt cccatgattc      360

cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg      420

taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt      480

ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt      540

atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacacctgc actcctccac      600

cttggctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg      660

aaaaagtggc accgagtcgg tgcttttttt acgcgta                               697


<210>  12
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX1sgRNA15T1-NNG

<400>  12
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa ctagaaactc gtagagtccc aggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccagag cctgcactcc      600

tccaccgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  13
<211>  7593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Plasmid 11254

<400>  13
ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc       60

attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga      120

gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt      180

gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt      240

gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg      300

acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca      360

aggccgcata tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg      420

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt      480

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca      540

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc      600

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta      660

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac      720

catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg      780

atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg      840

ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt      900

acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta      960

ccggtatggc ccctaagaag aagagaaaag tcggcggctc tggcagccag ctggtcaagt     1020

ctgagctgga agagaagaaa agcgagctgc ggcacaagct gaaatacgtg ccccacgagt     1080

acatcgagct gatcgagatc gccagaaaca gcacccagga cagaatcctg gaaatgaagg     1140

tcatggaatt ctttatgaag gtgtacggct accggggcaa gcacctcgga ggatctagaa     1200

agcctgacgg cgccatctat accgtgggca gccctatcga ttacggcgtg atcgtggata     1260

ccaaggccta cagcggcggc tacaatctgc ctattggaca ggccgacgag atgcagagat     1320

acgtggaaga aaaccagaca cggaacaagc acatcaaccc caacgagtgg tggaaagtgt     1380

accccagcag cgtgaccgag ttcaagttcc tgttcgtgtc cggccacttc aagggcaact     1440

acaaggccca gctgacccgg ctgaaccaca tcacaaattg caacggcgct gtgctgtctg     1500

tggaagaact gctgatcggc ggcgagatga tcaaggccgg aacactgaca ctggaagaag     1560

tgcggcggaa gttcaacaac ggcgagatca acttcagcgg cagcgagaca cctggcacat     1620

ctgaatctgc cacacctgag agcatggaaa agaagtacag catcggcctg gccatcggca     1680

caaattctgt tggctgggcc gtgatcaccg acgactacaa ggtgcccagc aagaaattca     1740

aggtgctggg caacaccaac cggaagtcca tcaagaaaaa cctgatgggc gccctgctgt     1800

tcgattctgg cgaaacagcc gaggccacca gactgaagag aaccgccaga agaagataca     1860

cccggcggaa gaaccggatc agatacctgc aagagatctt cgccaacgag atggccaagc     1920

tggacgacag cttctttcag agactggaag agtccttcct ggtggaagag gacaagaaga     1980

acgagcggca ccccatcttc ggcaatctgg ctgatgaggt ggcctaccac agaaactacc     2040

ccaccatcta ccacctgagg aagaagctgg ccgactctcc tgagaaggcc gacctgagac     2100

tgatctatct ggccctggct cacatcatca agttccgggg ccacttcctg atcgagggca     2160

agctgaatgc cgagaacagc gacgtggcca aactgttcta ccagctgatc cagacctaca     2220

accagctgtt cgaggaaagc cctctggacg agattgaggt ggacgccaag ggaatcctga     2280

gcgccagact gagcaagagc aagcggctgg aaaagctgat cgccgtgttt cccaacgaga     2340

agaagaatgg cctgttcggc aacatcattg ccctggcact gggcctgaca cctaacttca     2400

agtccaactt cgacctgacc gaggacgcca aactgcagct gagcaaggac acctacgacg     2460

acgatctgga tgagctgctg ggccagatcg gagatcagta cgccgatctg ttcagcgccg     2520

ccaagaatct gtctgacgcc atcctgctga gcgacatcct gagatccaac agcgaagtga     2580

caaaggcccc tctgagcgcc tccatggtca agagatatga cgagcaccac caggacctgg     2640

ctctgctgaa aacccttgtg cggcagcagt tccccgagaa gtatgccgag atcttcaagg     2700

acgacaccaa gaacggctac gccggctatg ttggcatcgg catcaagcac agaaagcgga     2760

ccaccaagct ggccacacaa gaggaattct acaagttcat caagcccatc ctcgagaaga     2820

tggacggcgc cgaagaactc ctggctaagc tgaacaggga cgacctgctg cggaagcaga     2880

gaaccttcga caatggcagc atccctcacc agatccacct gaaagagctg cacgccattc     2940

tgcggcggca agaagagttc tacccattcc tgaaagaaaa ccgggaaaag attgagaaga     3000

tcctgacctt caggatcccc tactacgtgg gaccactggc cagaggcaat agcagattcg     3060

cttggctgac cagaaagagc gaggaagcca tcacaccctg gaacttcgag gaagtggtgg     3120

ataagggcgc cagcgctcag tccttcatcg agcggatgac caatttcgac gagcagctgc     3180

ccaacaagaa ggtgctgcct aagcacagcc tgctgtacga gtacttcacc gtgtacaatg     3240

agctgaccaa agtcaaatac gtgacagagc ggatgcggaa gcctgagttt ctgagcggcg     3300

agcagaaaaa ggccatcgtg gatctgctgt tcaagaccaa cagaaaagtg accgtgaagc     3360

agctgaaaga ggactacttc aagaaaatcg agtgcttcga cagcgtcgag atcatcggcg     3420

tggaagatag attcaacgcc agcctgggca cataccacga tctgctgaag attatcaagg     3480

acaaggactt cctggacaac gaggaaaacg aggacatcct ggaagatatc gtgctgaccc     3540

tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacatac gcccacctgt     3600

tcgacgacaa agtgatgaag caactgaagc ggcggcacta cacaggctgg ggcagactgt     3660

ccagaaagat gattaacggc atccgggaca agcagagcgg caagaccatc ctggactttc     3720

tgaagtccga cggcttcagc aaccggaact tcatgcagct gattcacgac gacagcctca     3780

ccttcaaaga ggaaatcgag aaggcccagg tttccggcca gggcgattct ctgcacgagc     3840

agattgctga tctggccgga tctcccgcca ttaagaaggg catcctgcag acagtgaaga     3900

tcgtggacga actcgtgaaa gtgatgggcc acaagcctga gaacatcgtg atcgaaatgg     3960

ccagagagaa ccagaccacc accaagggcc tgcagcagtc cagagagaga aagaagcgcg     4020

gcggactgtc tgaagctgat aaggccggct tcattaagcg gcagctggtg gaaacccggc     4080

agatcacaaa gcacgtggcc cggatcctgg acagcagaat gaacaccaag cgggacaaga     4140

acgacaagcc tatccgggaa gtgaaagtca tcaccctgaa gtccaagctg gtgtccgatt     4200

tccggaagga tttccagctg tacaaagtgc gcgacatcaa caactaccat cacgcccacg     4260

acgcctacct gaatgctgtt gtgggaacag ccctgatcaa gaagtaccct aagctggaat     4320

ccgagttcgt gtacggcgac tacaaagtgt acgacgtgcg gaagatgatc gccaagagcg     4380

agcaagagat tggcaaggcc accgccaagc ggttcttcta ctccaacatc atgaactttt     4440

tcaagaccga agtgaagctc gccaacggcg aaatccggaa gaggcccctg attgagacaa     4500

atggcgagac aggcgaggtc gtgtggaaca aagagaagga cttcgccacc gtgcggaaag     4560

tgctggccat gcctcaagtg aatatcgtga aaaagaccga ggtgcagacc ggcggcttct     4620

ccaaagagag catcctgtct aagagagaga gcgccaagct catccctcgg aagaaaggct     4680

gggatacacg gaagtacggc ggatttggaa gccctaccgt ggcctactct attctggtgg     4740

tggccaaggt ggaaaagggc aaagccaaga aactgaagtc tgtgaaggtg ctcgtgggga     4800

tcaccatcat ggaaaaaggc agctacgaga aggaccctat cggctttctg gaagccaagg     4860

gctacaagga catcaaaaaa gagctgatct ttaagctccc caagtacagc ctgttcgaac     4920

tggaaaacgg cagacgcaga atgctggcct ctgccacaga actgcagaaa gccaatgaac     4980

tggtgctgcc acaacacctc gtgcggctgc tgtactacac ccagaatatc agcgccacca     5040

ccggcagcaa caacctggga tatatcgagc agcaccggga agagttcaaa gagattttcg     5100

agaaaatcat cgacttcagc gagaagtaca tcctgaagaa caaagtgaac agcaacctca     5160

agagcagctt cgatgagcag ttcgccgtgt ccgactctat cctgctgtct aacagcttcg     5220

tgtccctgct gaagtacacc tcctttggcg cctctggcgg cttcaccttt ctggatctgg     5280

acgtgaaaca ggggagactg agataccaga ccgtgactga ggtgctggac gccactctga     5340

tctaccagag catcaccggc ctgtacgaga caagaaccga tctctctcag ctcggaggcg     5400

actaaaagct tctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc     5460

ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc     5520

atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa     5580

gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggacaa     5640

acccacaact acgcgtactt gcggccgcat attcctagga tattctgggc ctcatgggcc     5700

ttcctttcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaacatggt     5760

catagctgtt tccttgcgta ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc     5820

tcggtcgttc gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa aaggccagga     5880

accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc     5940

acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg     6000

cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat     6060

acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt     6120

atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc     6180

agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg     6240

acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg     6300

gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg     6360

gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg     6420

gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca     6480

gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga     6540

acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga     6600

tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt     6660

ctgacagtta ttagaaaaat tcatccagca gacgataaaa cgcaatacgc tggctatccg     6720

gtgccgcaat gccatacagc accagaaaac gatccgccca ttcgccgccc agttcttccg     6780

caatatcacg ggtggccagc gcaatatcct gataacgatc cgccacgccc agacggccgc     6840

aatcaataaa gccgctaaaa cggccatttt ccaccataat gttcggcagg cacgcatcac     6900

catgggtcac caccagatct tcgccatccg gcatgctcgc tttcagacgc gcaaacagct     6960

ctgccggtgc caggccctga tgttcttcat ccagatcatc ctgatccacc aggcccgctt     7020

ccatacgggt acgcgcacgt tcaatacgat gtttcgcctg atgatcaaac ggacaggtcg     7080

ccgggtccag ggtatgcaga cgacgcatgg catccgccat aatgctcact ttttctgccg     7140

gcgccagatg gctagacagc agatcctgac ccggcacttc gcccagcagc agccaatcac     7200

ggcccgcttc ggtcaccaca tccagcaccg ccgcacacgg aacaccggtg gtggccagcc     7260

agctcagacg cgccgcttca tcctgcagct cgttcagcgc accgctcaga tcggttttca     7320

caaacagcac cggacgaccc tgcgcgctca gacgaaacac cgccgcatca gagcagccaa     7380

tggtctgctg cgcccaatca tagccaaaca gacgttccac ccacgctgcc gggctacccg     7440

catgcaggcc atcctgttca atcatactct tcctttttca atattattga agcatttatc     7500

agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag     7560

gggttccgcg cacatttccc cgaaaagtgc cac                                  7593


<210>  14
<211>  1353
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI.dHNH.dREC2

<400>  14

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Gly Gly Ser Gly Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala 
                405                 410                 415     


Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu 
            420                 425                 430         


Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe 
        435                 440                 445             


Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile 
    450                 455                 460                 


Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr 
465                 470                 475                 480 


Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu 
                485                 490                 495     


Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe 
            500                 505                 510         


Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala 
        515                 520                 525             


Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg 
    530                 535                 540                 


Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly 
545                 550                 555                 560 


Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser 
                565                 570                 575     


Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly 
            580                 585                 590         


Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln 
        595                 600                 605             


Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr 
    610                 615                 620                 


Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg 
625                 630                 635                 640 


Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val 
                645                 650                 655     


Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys 
            660                 665                 670         


Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile 
        675                 680                 685             


Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu 
    690                 695                 700                 


Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu 
705                 710                 715                 720 


Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg 
                725                 730                 735     


Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp 
            740                 745                 750         


Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg 
        755                 760                 765             


Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys 
    770                 775                 780                 


Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe 
785                 790                 795                 800 


Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu 
                805                 810                 815     


Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala 
            820                 825                 830         


Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val 
        835                 840                 845             


Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn 
    850                 855                 860                 


Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu 
865                 870                 875                 880 


Gln Gln Ser Arg Glu Arg Lys Lys Arg Gly Gly Ser Gly Gly Leu Ser 
                885                 890                 895     


Glu Ala Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg 
            900                 905                 910         


Gln Ile Thr Lys His Val Ala Arg Ile Leu Asp Ser Arg Met Asn Thr 
        915                 920                 925             


Lys Arg Asp Lys Asn Asp Lys Pro Ile Arg Glu Val Lys Val Ile Thr 
    930                 935                 940                 


Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Leu Tyr 
945                 950                 955                 960 


Lys Val Arg Asp Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu 
                965                 970                 975     


Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu 
            980                 985                 990         


Ser Glu Phe Val Tyr Gly Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met 
        995                 1000                 1005             


Ile Ala  Lys Ser Glu Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg 
    1010                 1015                 1020             


Phe Phe  Tyr Ser Asn Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys 
    1025                 1030                 1035             


Leu Ala  Asn Gly Glu Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn 
    1040                 1045                 1050             


Gly Glu  Thr Gly Glu Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala 
    1055                 1060                 1065             


Thr Val  Arg Lys Val Leu Ala  Met Pro Gln Val Asn  Ile Val Lys 
    1070                 1075                 1080             


Lys Thr  Glu Val Gln Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu 
    1085                 1090                 1095             


Ser Lys  Arg Glu Ser Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp 
    1100                 1105                 1110             


Asp Thr  Arg Lys Tyr Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr 
    1115                 1120                 1125             


Ser Ile  Leu Val Val Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys 
    1130                 1135                 1140             


Leu Lys  Ser Val Lys Val Leu  Val Gly Ile Thr Ile  Met Glu Lys 
    1145                 1150                 1155             


Gly Ser  Tyr Glu Lys Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly 
    1160                 1165                 1170             


Tyr Lys  Asp Ile Lys Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr 
    1175                 1180                 1185             


Ser Leu  Phe Glu Leu Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser 
    1190                 1195                 1200             


Ala Thr  Glu Leu Gln Lys Ala  Asn Glu Leu Val Leu  Pro Gln His 
    1205                 1210                 1215             


Leu Val  Arg Leu Leu Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr 
    1220                 1225                 1230             


Gly Ser  Asn Asn Leu Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe 
    1235                 1240                 1245             


Lys Glu  Ile Phe Glu Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile 
    1250                 1255                 1260             


Leu Lys  Asn Lys Val Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu 
    1265                 1270                 1275             


Gln Phe  Ala Val Ser Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val 
    1280                 1285                 1290             


Ser Leu  Leu Lys Tyr Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr 
    1295                 1300                 1305             


Phe Leu  Asp Leu Asp Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr 
    1310                 1315                 1320             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1325                 1330                 1335             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1340                 1345                 1350             


<210>  15
<211>  1424
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI.dHNH.dFLEX

<400>  15

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
                805                 810                 815     


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
            820                 825                 830         


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
        835                 840                 845             


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
    850                 855                 860                 


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
865                 870                 875                 880 


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
                885                 890                 895     


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
            900                 905                 910         


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
        915                 920                 925             


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
    930                 935                 940                 


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
945                 950                 955                 960 


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
                965                 970                 975     


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
            980                 985                 990         


Asn Ile Val Ile Glu Met Ala Arg  Glu Asn Gln Thr Thr  Thr Lys Gly 
        995                 1000                 1005             


Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg Gly Gly  Ser Gly Gly 
    1010                 1015                 1020             


Leu Ser  Glu Ala Asp Lys Ala  Gly Phe Ile Lys Arg  Gln Leu Val 
    1025                 1030                 1035             


Glu Thr  Arg Gln Ile Thr Lys  His Val Ala Arg Ile  Leu Asp Ser 
    1040                 1045                 1050             


Arg Met  Asn Thr Lys Arg Asp  Lys Asn Asp Lys Pro  Ile Arg Glu 
    1055                 1060                 1065             


Val Lys  Val Ile Thr Leu Lys  Ser Lys Leu Val Ser  Asp Phe Arg 
    1070                 1075                 1080             


Lys Asp  Phe Gln Leu Tyr Lys  Val Arg Asp Ile Asn  Asn Tyr His 
    1085                 1090                 1095             


His Ala  His Asp Ala Tyr Leu  Asn Ala Val Val Gly  Thr Ala Leu 
    1100                 1105                 1110             


Ile Lys  Lys Tyr Pro Lys Leu  Glu Ser Glu Phe Val  Tyr Gly Gly 
    1115                 1120                 1125             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1130                 1135                 1140             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1145                 1150                 1155             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1160                 1165                 1170             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1175                 1180                 1185             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1190                 1195                 1200             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1205                 1210                 1215             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1220                 1225                 1230             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1235                 1240                 1245             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1250                 1255                 1260             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1265                 1270                 1275             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1280                 1285                 1290             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1295                 1300                 1305             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1310                 1315                 1320             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1325                 1330                 1335             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1340                 1345                 1350             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1355                 1360                 1365             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1370                 1375                 1380             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1385                 1390                 1395             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1400                 1405                 1410             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp 
    1415                 1420                 


<210>  16
<211>  1468
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI.dREC2.dFLEX

<400>  16

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala Ser Met Val Lys 
                405                 410                 415     


Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr Leu Val 
            420                 425                 430         


Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp Asp Thr 
        435                 440                 445             


Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His Arg Lys 
    450                 455                 460                 


Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
465                 470                 475                 480 


Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu 
                485                 490                 495     


Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
            500                 505                 510         


Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu Arg Arg 
        515                 520                 525             


Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys Ile Glu 
    530                 535                 540                 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
545                 550                 555                 560 


Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu Ala Ile 
                565                 570                 575     


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
            580                 585                 590         


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
        595                 600                 605             


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
    610                 615                 620                 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
625                 630                 635                 640 


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
                645                 650                 655     


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
            660                 665                 670         


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
        675                 680                 685             


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
    690                 695                 700                 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
705                 710                 715                 720 


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
                725                 730                 735     


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
            740                 745                 750         


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
        755                 760                 765             


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
    770                 775                 780                 


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
785                 790                 795                 800 


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
                805                 810                 815     


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly 
            820                 825                 830         


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp 
        835                 840                 845             


Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu 
    850                 855                 860                 


Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu Gln Gln Ser Arg 
865                 870                 875                 880 


Glu Arg Lys Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Glu Ser Gln 
                885                 890                 895     


Ile Leu Lys Glu Asn Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys 
            900                 905                 910         


Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln 
        915                 920                 925             


Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile Val 
    930                 935                 940                 


Pro Gln Ser Phe Ile Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr 
945                 950                 955                 960 


Arg Ser Val Glu Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu 
                965                 970                 975     


Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys 
            980                 985                 990         


Leu Ile Thr Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
        995                 1000                 1005             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1010                 1015                 1020             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1025                 1030                 1035             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1040                 1045                 1050             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1055                 1060                 1065             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1070                 1075                 1080             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1085                 1090                 1095             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1100                 1105                 1110             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1115                 1120                 1125             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1130                 1135                 1140             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1145                 1150                 1155             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1160                 1165                 1170             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1175                 1180                 1185             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1190                 1195                 1200             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1205                 1210                 1215             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1220                 1225                 1230             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1235                 1240                 1245             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1250                 1255                 1260             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1265                 1270                 1275             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1280                 1285                 1290             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1295                 1300                 1305             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1310                 1315                 1320             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1325                 1330                 1335             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1340                 1345                 1350             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1355                 1360                 1365             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1370                 1375                 1380             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1385                 1390                 1395             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1400                 1405                 1410             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1415                 1420                 1425             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1430                 1435                 1440             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1445                 1450                 1455             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1460                 1465             


<210>  17
<211>  1295
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI.dHNH.dREC2.dFLEX

<400>  17

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Gly Gly Ser Gly Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala 
                405                 410                 415     


Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu 
            420                 425                 430         


Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe 
        435                 440                 445             


Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile 
    450                 455                 460                 


Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr 
465                 470                 475                 480 


Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu 
                485                 490                 495     


Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe 
            500                 505                 510         


Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala 
        515                 520                 525             


Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg 
    530                 535                 540                 


Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly 
545                 550                 555                 560 


Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser 
                565                 570                 575     


Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly 
            580                 585                 590         


Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln 
        595                 600                 605             


Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr 
    610                 615                 620                 


Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg 
625                 630                 635                 640 


Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val 
                645                 650                 655     


Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys 
            660                 665                 670         


Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile 
        675                 680                 685             


Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu 
    690                 695                 700                 


Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu 
705                 710                 715                 720 


Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg 
                725                 730                 735     


Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp 
            740                 745                 750         


Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg 
        755                 760                 765             


Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys 
    770                 775                 780                 


Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe 
785                 790                 795                 800 


Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu 
                805                 810                 815     


Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala 
            820                 825                 830         


Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val 
        835                 840                 845             


Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn 
    850                 855                 860                 


Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu 
865                 870                 875                 880 


Gln Gln Ser Arg Glu Arg Lys Lys Arg Gly Gly Ser Gly Gly Leu Ser 
                885                 890                 895     


Glu Ala Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg 
            900                 905                 910         


Gln Ile Thr Lys His Val Ala Arg Ile Leu Asp Ser Arg Met Asn Thr 
        915                 920                 925             


Lys Arg Asp Lys Asn Asp Lys Pro Ile Arg Glu Val Lys Val Ile Thr 
    930                 935                 940                 


Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Leu Tyr 
945                 950                 955                 960 


Lys Val Arg Asp Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu 
                965                 970                 975     


Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu 
            980                 985                 990         


Ser Glu Phe Val Tyr Gly Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp 
        995                 1000                 1005             


Phe Ala  Thr Val Arg Lys Val  Leu Ala Met Pro Gln  Val Asn Ile 
    1010                 1015                 1020             


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1025                 1030                 1035             


Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys 
    1040                 1045                 1050             


Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val 
    1055                 1060                 1065             


Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ala 
    1070                 1075                 1080             


Lys Lys  Leu Lys Ser Val Lys  Val Leu Val Gly Ile  Thr Ile Met 
    1085                 1090                 1095             


Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala 
    1100                 1105                 1110             


Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro 
    1115                 1120                 1125             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu 
    1130                 1135                 1140             


Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro 
    1145                 1150                 1155             


Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala 
    1160                 1165                 1170             


Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu 
    1175                 1180                 1185             


Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys 
    1190                 1195                 1200             


Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe 
    1205                 1210                 1215             


Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser 
    1220                 1225                 1230             


Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly 
    1235                 1240                 1245             


Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr 
    1250                 1255                 1260             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1265                 1270                 1275             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1280                 1285                 1290             


Gly Asp  
    1295 


<210>  18
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3042-EMX1-T1-ddPCR-F1

<400>  18
gaaaccatgc ccattc                                                       16


<210>  19
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3049-EMX1-T1-ddPCR-R2

<400>  19
agagaagggt ggttttc                                                      17


<210>  20
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3088-EMX1-T1-TGEE3-reference probe


<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  g is modified with ZEN

<400>  20
ctctgtatgg aaaagagcat                                                   20


<210>  21
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3089-EMX1-T1-TGEE3-drop off probe


<220>
<221>  misc_feature
<222>  (8)..(8)
<223>  g is a locked nucleic acid

<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  t is a locked nucleic acid

<400>  21
tcgtagagtc ccatgtc                                                      17


<210>  22
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3175-EMX1-T1-TGEE4-drop off probe


<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  c is a locked nucleic acid

<220>
<221>  misc_feature
<222>  (10)..(10)
<223>  g is a locked nucleic acid

<400>  22
atgtctgccg gcttcc                                                       16


<210>  23
<211>  3027
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Plasmid 10917

<400>  23
ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc       60

attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga      120

gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt      180

gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt      240

gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg      300

acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca      360

aggccgcata ccggttggac aaacccacaa ctacgcgtaa aaaaaaagca ccgactcggt      420

gccacttttt caagttgata acggactagc cttattttaa cttgctattt ctagctctaa      480

aactagaaac tcgtagagtc ccaggtgttt cgtggtctca tacagaactt ataagattcc      540

caaatccaaa gacatttcac gtttatggtg atttcccaga acacatagcg acatgcaaat      600

attgcagggc gccactcccc tgtccctcac agccatcttc ctgccagggc gcacgcgcgc      660

tgggtgttcc cgcctagtga cactgggccc gcgattcctt ggagcgggtt gatgacgtca      720

gcgttcaaag agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct      780

gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg      840

tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg      900

gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg      960

tggaaaggac gaaacaccag agcctgcact cctccaccgt tttagagcta gaaatagcaa     1020

gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt     1080

tacgcgtact tgcggccgca tattcctagg ataccggtct gggcctcatg ggccttcctt     1140

tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaaca tggtcatagc     1200

tgtttccttg cgtattgggc gctctccgct tcctcgctca ctgactcgct gcgctcggtc     1260

gttcgggtaa agcctggggt gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta     1320

aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa     1380

atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc     1440

cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt     1500

ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca     1560

gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg     1620

accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat     1680

cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta     1740

cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct     1800

gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac     1860

aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa     1920

aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa     1980

actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt     2040

taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca     2100

gttattagaa aaattcatcc agcagacgat aaaacgcaat acgctggcta tccggtgccg     2160

caatgccata cagcaccaga aaacgatccg cccattcgcc gcccagttct tccgcaatat     2220

cacgggtggc cagcgcaata tcctgataac gatccgccac gcccagacgg ccgcaatcaa     2280

taaagccgct aaaacggcca ttttccacca taatgttcgg caggcacgca tcaccatggg     2340

tcaccaccag atcttcgcca tccggcatgc tcgctttcag acgcgcaaac agctctgccg     2400

gtgccaggcc ctgatgttct tcatccagat catcctgatc caccaggccc gcttccatac     2460

gggtacgcgc acgttcaata cgatgtttcg cctgatgatc aaacggacag gtcgccgggt     2520

ccagggtatg cagacgacgc atggcatccg ccataatgct cactttttct gccggcgcca     2580

gatggctaga cagcagatcc tgacccggca cttcgcccag cagcagccaa tcacggcccg     2640

cttcggtcac cacatccagc accgccgcac acggaacacc ggtggtggcc agccagctca     2700

gacgcgccgc ttcatcctgc agctcgttca gcgcaccgct cagatcggtt ttcacaaaca     2760

gcaccggacg accctgcgcg ctcagacgaa acaccgccgc atcagagcag ccaatggtct     2820

gctgcgccca atcatagcca aacagacgtt ccacccacgc tgccgggcta cccgcatgca     2880

ggccatcctg ttcaatcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt     2940

attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc     3000

cgcgcacatt tccccgaaaa gtgccac                                         3027


<210>  24
<211>  1537
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-ZF1

<400>  24

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1445                 1450                 1455             


Ser Ala  Pro Tyr Arg Cys Pro  Cys Arg Ala Gly Cys  Pro Ser Leu 
    1460                 1465                 1470             


Ala Ser  Met Gln Ala His Met  Arg Gly His Ser Gly  Gly Ser Gly 
    1475                 1480                 1485             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1490                 1495                 1500             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1505                 1510                 1515             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1520                 1525                 1530             


Lys Lys  Lys Lys 
    1535         


<210>  25
<211>  1630
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-ZF2

<400>  25

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Gly 
    1535                 1540                 1545             


Thr Gly  Gly Ser Ala Pro Tyr  Arg Cys Pro Cys Arg  Ala Gly Cys 
    1550                 1555                 1560             


Pro Ser  Leu Ala Ser Met Gln  Ala His Met Arg Gly  His Ser Gly 
    1565                 1570                 1575             


Gly Ser  Gly Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1580                 1585                 1590             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1595                 1600                 1605             


Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala 
    1610                 1615                 1620             


Gly Gln  Ala Lys Lys Lys Lys  
    1625                 1630 


<210>  26
<211>  1542
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-ZF3

<400>  26

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1445                 1450                 1455             


Ser Ala  Gly Gly Ser Gly Ala  Pro Tyr Arg Cys Pro  Cys Arg Ala 
    1460                 1465                 1470             


Gly Cys  Pro Ser Leu Ala Ser  Met Gln Ala His Met  Arg Gly His 
    1475                 1480                 1485             


Ser Gly  Gly Ser Gly Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1490                 1495                 1500             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1505                 1510                 1515             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1520                 1525                 1530             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1535                 1540         


<210>  27
<211>  1635
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-ZF4

<400>  27

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Gly 
    1535                 1540                 1545             


Thr Gly  Gly Ser Ala Gly Gly  Ser Gly Ala Pro Tyr  Arg Cys Pro 
    1550                 1555                 1560             


Cys Arg  Ala Gly Cys Pro Ser  Leu Ala Ser Met Gln  Ala His Met 
    1565                 1570                 1575             


Arg Gly  His Ser Gly Gly Ser  Gly Tyr Gln Thr Val  Thr Glu Val 
    1580                 1585                 1590             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1595                 1600                 1605             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala 
    1610                 1615                 1620             


Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys Lys Lys  
    1625                 1630                 1635 


<210>  28
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-LAC

<400>  28

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Gly Met Lys  Pro Val Thr 
    1445                 1450                 1455             


Leu Tyr  Asp Val Ala Glu Tyr  Ala Gly Val Ser Tyr  Gln Thr Val 
    1460                 1465                 1470             


Ser Arg  Val Val Asn Gln Ala  Ser His Val Ser Ala  Lys Thr Arg 
    1475                 1480                 1485             


Glu Lys  Val Glu Ala Ala Met  Ala Glu Leu Asn Gly  Gly Ser Gly 
    1490                 1495                 1500             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1505                 1510                 1515             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1520                 1525                 1530             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1535                 1540                 1545             


Lys Lys  Lys Lys 
    1550         


<210>  29
<211>  1645
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-LAC2

<400>  29

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Gly Met Lys 
    1535                 1540                 1545             


Pro Val  Thr Leu Tyr Asp Val  Ala Glu Tyr Ala Gly  Val Ser Tyr 
    1550                 1555                 1560             


Gln Thr  Val Ser Arg Val Val  Asn Gln Ala Ser His  Val Ser Ala 
    1565                 1570                 1575             


Lys Thr  Arg Glu Lys Val Glu  Ala Ala Met Ala Glu  Leu Asn Gly 
    1580                 1585                 1590             


Gly Ser  Gly Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1595                 1600                 1605             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1610                 1615                 1620             


Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala 
    1625                 1630                 1635             


Gly Gln  Ala Lys Lys Lys Lys  
    1640                 1645 


<210>  30
<211>  1564
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-LAC3

<400>  30

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1445                 1450                 1455             


Ser Met  Lys Pro Val Thr Leu  Tyr Asp Val Ala Glu  Tyr Ala Gly 
    1460                 1465                 1470             


Val Ser  Tyr Gln Thr Val Ser  Arg Val Val Asn Gln  Ala Ser His 
    1475                 1480                 1485             


Val Ser  Ala Lys Thr Arg Glu  Lys Val Glu Ala Ala  Met Ala Glu 
    1490                 1495                 1500             


Leu Asn  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Tyr Gln Thr 
    1505                 1510                 1515             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1520                 1525                 1530             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1535                 1540                 1545             


Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys 
    1550                 1555                 1560             


Lys 
    


<210>  31
<211>  1657
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-LAC4

<400>  31

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Gly 
    1535                 1540                 1545             


Thr Gly  Gly Ser Met Lys Pro  Val Thr Leu Tyr Asp  Val Ala Glu 
    1550                 1555                 1560             


Tyr Ala  Gly Val Ser Tyr Gln  Thr Val Ser Arg Val  Val Asn Gln 
    1565                 1570                 1575             


Ala Ser  His Val Ser Ala Lys  Thr Arg Glu Lys Val  Glu Ala Ala 
    1580                 1585                 1590             


Met Ala  Glu Leu Asn Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser 
    1595                 1600                 1605             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1610                 1615                 1620             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1625                 1630                 1635             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1640                 1645                 1650             


Lys Lys  Lys Lys 
    1655         


<210>  32
<211>  1555
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-HIVIN

<400>  32

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Gly Ile Gln  Asn Phe Arg 
    1445                 1450                 1455             


Val Tyr  Tyr Arg Asp Ser Arg  Asn Pro Leu Trp Lys  Gly Pro Ala 
    1460                 1465                 1470             


Lys Leu  Leu Trp Lys Gly Glu  Gly Ala Trp Ile Gln  Asp Asn Ser 
    1475                 1480                 1485             


Asp Ile  Lys Trp Pro Arg Arg  Lys Ala Lys Ile Ile  Arg Asp Gly 
    1490                 1495                 1500             


Gly Ser  Gly Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1505                 1510                 1515             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1520                 1525                 1530             


Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala 
    1535                 1540                 1545             


Gly Gln  Ala Lys Lys Lys Lys  
    1550                 1555 


<210>  33
<211>  1648
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-HIVIN2

<400>  33

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Gly Ile Gln 
    1535                 1540                 1545             


Asn Phe  Arg Val Tyr Tyr Arg  Asp Ser Arg Asn Pro  Leu Trp Lys 
    1550                 1555                 1560             


Gly Pro  Ala Lys Leu Leu Trp  Lys Gly Glu Gly Ala  Trp Ile Gln 
    1565                 1570                 1575             


Asp Asn  Ser Asp Ile Lys Trp  Pro Arg Arg Lys Ala  Lys Ile Ile 
    1580                 1585                 1590             


Arg Asp  Gly Gly Ser Gly Tyr  Gln Thr Val Thr Glu  Val Leu Asp 
    1595                 1600                 1605             


Ala Thr  Leu Ile Tyr Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1610                 1615                 1620             


Thr Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1625                 1630                 1635             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys 
    1640                 1645             


<210>  34
<211>  1567
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-HIVIN3

<400>  34

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1445                 1450                 1455             


Ser Ile  Gln Asn Phe Arg Val  Tyr Tyr Arg Asp Ser  Arg Asn Pro 
    1460                 1465                 1470             


Leu Trp  Lys Gly Pro Ala Lys  Leu Leu Trp Lys Gly  Glu Gly Ala 
    1475                 1480                 1485             


Trp Ile  Gln Asp Asn Ser Asp  Ile Lys Trp Pro Arg  Arg Lys Ala 
    1490                 1495                 1500             


Lys Ile  Ile Arg Asp Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser 
    1505                 1510                 1515             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1520                 1525                 1530             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1535                 1540                 1545             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1550                 1555                 1560             


Lys Lys  Lys Lys 
    1565         


<210>  35
<211>  1660
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-HIVIN4

<400>  35

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Gly 
    1535                 1540                 1545             


Thr Gly  Gly Ser Ile Gln Asn  Phe Arg Val Tyr Tyr  Arg Asp Ser 
    1550                 1555                 1560             


Arg Asn  Pro Leu Trp Lys Gly  Pro Ala Lys Leu Leu  Trp Lys Gly 
    1565                 1570                 1575             


Glu Gly  Ala Trp Ile Gln Asp  Asn Ser Asp Ile Lys  Trp Pro Arg 
    1580                 1585                 1590             


Arg Lys  Ala Lys Ile Ile Arg  Asp Gly Gly Ser Ala  Gly Gly Thr 
    1595                 1600                 1605             


Gly Gly  Ser Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1610                 1615                 1620             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1625                 1630                 1635             


Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala 
    1640                 1645                 1650             


Gly Gln  Ala Lys Lys Lys Lys  
    1655                 1660 


<210>  36
<211>  1572
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-SSO7D

<400>  36

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Met  Ala Thr Val 
    1445                 1450                 1455             


Lys Phe  Lys Tyr Lys Gly Glu  Glu Lys Glu Val Asp  Ile Ser Lys 
    1460                 1465                 1470             


Ile Lys  Lys Val Trp Arg Val  Gly Lys Met Ile Ser  Phe Thr Tyr 
    1475                 1480                 1485             


Asp Glu  Gly Gly Gly Lys Thr  Gly Arg Gly Ala Val  Ser Glu Lys 
    1490                 1495                 1500             


Asp Ala  Pro Lys Glu Leu Leu  Gln Met Leu Glu Lys  Gln Lys Lys 
    1505                 1510                 1515             


Gly Gly  Ser Ala Gly Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1520                 1525                 1530             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1535                 1540                 1545             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1550                 1555                 1560             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1565                 1570         


<210>  37
<211>  1665
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-SSO7D2

<400>  37

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Met 
    1535                 1540                 1545             


Ala Thr  Val Lys Phe Lys Tyr  Lys Gly Glu Glu Lys  Glu Val Asp 
    1550                 1555                 1560             


Ile Ser  Lys Ile Lys Lys Val  Trp Arg Val Gly Lys  Met Ile Ser 
    1565                 1570                 1575             


Phe Thr  Tyr Asp Glu Gly Gly  Gly Lys Thr Gly Arg  Gly Ala Val 
    1580                 1585                 1590             


Ser Glu  Lys Asp Ala Pro Lys  Glu Leu Leu Gln Met  Leu Glu Lys 
    1595                 1600                 1605             


Gln Lys  Lys Gly Gly Ser Ala  Gly Tyr Gln Thr Val  Thr Glu Val 
    1610                 1615                 1620             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1625                 1630                 1635             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala 
    1640                 1645                 1650             


Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys Lys Lys  
    1655                 1660                 1665 


<210>  38
<211>  1582
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-SSO7D3

<400>  38

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1445                 1450                 1455             


Ser Met  Ala Thr Val Lys Phe  Lys Tyr Lys Gly Glu  Glu Lys Glu 
    1460                 1465                 1470             


Val Asp  Ile Ser Lys Ile Lys  Lys Val Trp Arg Val  Gly Lys Met 
    1475                 1480                 1485             


Ile Ser  Phe Thr Tyr Asp Glu  Gly Gly Gly Lys Thr  Gly Arg Gly 
    1490                 1495                 1500             


Ala Val  Ser Glu Lys Asp Ala  Pro Lys Glu Leu Leu  Gln Met Leu 
    1505                 1510                 1515             


Glu Lys  Gln Lys Lys Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser 
    1520                 1525                 1530             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1535                 1540                 1545             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1550                 1555                 1560             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1565                 1570                 1575             


Lys Lys  Lys Lys 
    1580         


<210>  39
<211>  1675
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-SSO7D4

<400>  39

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Gly 
    1535                 1540                 1545             


Thr Gly  Gly Ser Met Ala Thr  Val Lys Phe Lys Tyr  Lys Gly Glu 
    1550                 1555                 1560             


Glu Lys  Glu Val Asp Ile Ser  Lys Ile Lys Lys Val  Trp Arg Val 
    1565                 1570                 1575             


Gly Lys  Met Ile Ser Phe Thr  Tyr Asp Glu Gly Gly  Gly Lys Thr 
    1580                 1585                 1590             


Gly Arg  Gly Ala Val Ser Glu  Lys Asp Ala Pro Lys  Glu Leu Leu 
    1595                 1600                 1605             


Gln Met  Leu Glu Lys Gln Lys  Lys Gly Gly Ser Ala  Gly Gly Thr 
    1610                 1615                 1620             


Gly Gly  Ser Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1625                 1630                 1635             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1640                 1645                 1650             


Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala 
    1655                 1660                 1665             


Gly Gln  Ala Lys Lys Lys Lys  
    1670                 1675 


<210>  40
<211>  1572
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-STO7

<400>  40

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Met  Val Thr Val 
    1445                 1450                 1455             


Lys Phe  Lys Tyr Lys Gly Glu  Glu Lys Glu Val Asp  Ile Ser Lys 
    1460                 1465                 1470             


Ile Lys  Lys Val Trp Arg Val  Gly Lys Met Ile Ser  Phe Thr Tyr 
    1475                 1480                 1485             


Asp Asp  Asn Gly Lys Thr Gly  Arg Gly Ala Val Ser  Glu Lys Asp 
    1490                 1495                 1500             


Ala Pro  Lys Glu Leu Leu Gln  Met Leu Glu Lys Ser  Gly Lys Lys 
    1505                 1510                 1515             


Gly Gly  Ser Ala Gly Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1520                 1525                 1530             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1535                 1540                 1545             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1550                 1555                 1560             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1565                 1570         


<210>  41
<211>  1665
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-STO72

<400>  41

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Met 
    1535                 1540                 1545             


Val Thr  Val Lys Phe Lys Tyr  Lys Gly Glu Glu Lys  Glu Val Asp 
    1550                 1555                 1560             


Ile Ser  Lys Ile Lys Lys Val  Trp Arg Val Gly Lys  Met Ile Ser 
    1565                 1570                 1575             


Phe Thr  Tyr Asp Asp Asn Gly  Lys Thr Gly Arg Gly  Ala Val Ser 
    1580                 1585                 1590             


Glu Lys  Asp Ala Pro Lys Glu  Leu Leu Gln Met Leu  Glu Lys Ser 
    1595                 1600                 1605             


Gly Lys  Lys Gly Gly Ser Ala  Gly Tyr Gln Thr Val  Thr Glu Val 
    1610                 1615                 1620             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1625                 1630                 1635             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala 
    1640                 1645                 1650             


Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys Lys Lys  
    1655                 1660                 1665 


<210>  42
<211>  1582
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-STO73

<400>  42

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1445                 1450                 1455             


Ser Met  Val Thr Val Lys Phe  Lys Tyr Lys Gly Glu  Glu Lys Glu 
    1460                 1465                 1470             


Val Asp  Ile Ser Lys Ile Lys  Lys Val Trp Arg Val  Gly Lys Met 
    1475                 1480                 1485             


Ile Ser  Phe Thr Tyr Asp Asp  Asn Gly Lys Thr Gly  Arg Gly Ala 
    1490                 1495                 1500             


Val Ser  Glu Lys Asp Ala Pro  Lys Glu Leu Leu Gln  Met Leu Glu 
    1505                 1510                 1515             


Lys Ser  Gly Lys Lys Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser 
    1520                 1525                 1530             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1535                 1540                 1545             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1550                 1555                 1560             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1565                 1570                 1575             


Lys Lys  Lys Lys 
    1580         


<210>  43
<211>  1675
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-STO74

<400>  43

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
225                 230                 235                 240 


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
                245                 250                 255     


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
            260                 265                 270         


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
        275                 280                 285             


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
    290                 295                 300                 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
305                 310                 315                 320 


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
                325                 330                 335     


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
            340                 345                 350         


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
        355                 360                 365             


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
    370                 375                 380                 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
385                 390                 395                 400 


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
                405                 410                 415     


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
            420                 425                 430         


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
        435                 440                 445             


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
    450                 455                 460                 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
465                 470                 475                 480 


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
                485                 490                 495     


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
            500                 505                 510         


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
        515                 520                 525             


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Met 
    530                 535                 540                 


Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr 
545                 550                 555                 560 


Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp 
                565                 570                 575     


Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His 
            580                 585                 590         


Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe 
        595                 600                 605             


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala 
    610                 615                 620                 


Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
625                 630                 635                 640 


Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu 
                645                 650                 655     


Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys 
            660                 665                 670         


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
        675                 680                 685             


Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu 
    690                 695                 700                 


Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Gln 
705                 710                 715                 720 


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys 
                725                 730                 735     


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
            740                 745                 750         


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro 
        755                 760                 765             


Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
    770                 775                 780                 


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
785                 790                 795                 800 


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp 
                805                 810                 815     


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
            820                 825                 830         


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
        835                 840                 845             


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
    850                 855                 860                 


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
865                 870                 875                 880 


Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
                885                 890                 895     


Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
            900                 905                 910         


Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile 
        915                 920                 925             


His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val 
    930                 935                 940                 


Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Val Arg  Lys Val Leu Ala Met  Pro Gln Val Asn Ile  Val Lys Lys 
    1310                 1315                 1320             


Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser 
    1325                 1330                 1335             


Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp 
    1340                 1345                 1350             


Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser 
    1355                 1360                 1365             


Ile Leu  Val Val Ala Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu 
    1370                 1375                 1380             


Lys Ser  Val Lys Val Leu Val  Gly Ile Thr Ile Met  Glu Lys Gly 
    1385                 1390                 1395             


Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr 
    1400                 1405                 1410             


Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys Leu Pro  Lys Tyr Ser 
    1415                 1420                 1425             


Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala 
    1430                 1435                 1440             


Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val Leu Pro  Gln His Leu 
    1445                 1450                 1455             


Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile Ser Ala  Thr Thr Gly 
    1460                 1465                 1470             


Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His Arg Glu  Glu Phe Lys 
    1475                 1480                 1485             


Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu 
    1490                 1495                 1500             


Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser Ser Phe  Asp Glu Gln 
    1505                 1510                 1515             


Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser Asn Ser  Phe Val Ser 
    1520                 1525                 1530             


Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Gly Gly Ser  Ala Gly Gly 
    1535                 1540                 1545             


Thr Gly  Gly Ser Met Val Thr  Val Lys Phe Lys Tyr  Lys Gly Glu 
    1550                 1555                 1560             


Glu Lys  Glu Val Asp Ile Ser  Lys Ile Lys Lys Val  Trp Arg Val 
    1565                 1570                 1575             


Gly Lys  Met Ile Ser Phe Thr  Tyr Asp Asp Asn Gly  Lys Thr Gly 
    1580                 1585                 1590             


Arg Gly  Ala Val Ser Glu Lys  Asp Ala Pro Lys Glu  Leu Leu Gln 
    1595                 1600                 1605             


Met Leu  Glu Lys Ser Gly Lys  Lys Gly Gly Ser Ala  Gly Gly Thr 
    1610                 1615                 1620             


Gly Gly  Ser Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1625                 1630                 1635             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1640                 1645                 1650             


Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala 
    1655                 1660                 1665             


Gly Gln  Ala Lys Lys Lys Lys  
    1670                 1675 


<210>  44
<211>  1436
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-StkC

<400>  44

Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu Met 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Arg 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp Ser 
                85                  90                  95      


Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala Tyr 
        115                 120                 125             


His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp 
    130                 135                 140                 


Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn Ala 
                165                 170                 175     


Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp Ala 
        195                 200                 205             


Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu Lys 
    210                 215                 220                 


Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys 
                325                 330                 335     


Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys 
            340                 345                 350         


Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys 
        355                 360                 365             


His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys 
    370                 375                 380                 


Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu 
385                 390                 395                 400 


Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp 
                405                 410                 415     


Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile 
            420                 425                 430         


Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu 
        435                 440                 445             


Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro 
    450                 455                 460                 


Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu 
465                 470                 475                 480 


Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala 
                485                 490                 495     


Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu 
            500                 505                 510         


Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe 
        515                 520                 525             


Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met 
    530                 535                 540                 


Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp 
545                 550                 555                 560 


Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu 
                565                 570                 575     


Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly 
            580                 585                 590         


Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu 
        595                 600                 605             


Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp 
    610                 615                 620                 


Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu 
625                 630                 635                 640 


Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys 
                645                 650                 655     


Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu 
            660                 665                 670         


Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr 
        675                 680                 685             


Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met 
    690                 695                 700                 


Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys 
705                 710                 715                 720 


Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp 
                725                 730                 735     


Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys 
            740                 745                 750         


Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn Ile 
        755                 760                 765             


Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu Gln 
    770                 775                 780                 


Gln Ser Arg Glu Arg Lys Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu 
785                 790                 795                 800 


Glu Ser Gln Ile Leu Lys Glu Asn Pro Val Glu Asn Thr Gln Leu Gln 
                805                 810                 815     


Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr 
            820                 825                 830         


Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp 
        835                 840                 845             


Ala Ile Val Pro Gln Ser Phe Ile Lys Asp Asp Ser Ile Asp Asn Lys 
    850                 855                 860                 


Val Leu Thr Arg Ser Val Glu Asn Arg Gly Lys Ser Asp Asn Val Pro 
865                 870                 875                 880 


Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu 
                885                 890                 895     


Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala 
            900                 905                 910         


Glu Arg Gly Gly Leu Ser Glu Ala Asp Lys Ala Gly Phe Ile Lys Arg 
        915                 920                 925             


Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Arg Ile Leu 
    930                 935                 940                 


Asp Ser Arg Met Asn Thr Lys Arg Asp Lys Asn Asp Lys Pro Ile Arg 
945                 950                 955                 960 


Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg 
                965                 970                 975     


Lys Asp Phe Gln Leu Tyr Lys Val Arg Asp Ile Asn Asn Tyr His His 
            980                 985                 990         


Ala His Asp Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
        995                 1000                 1005             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1010                 1015                 1020             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1025                 1030                 1035             


Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1040                 1045                 1050             


Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1055                 1060                 1065             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp 
    1070                 1075                 1080             


Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg Lys Val  Leu Ala Met 
    1085                 1090                 1095             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1100                 1105                 1110             


Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu 
    1115                 1120                 1125             


Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe 
    1130                 1135                 1140             


Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val 
    1145                 1150                 1155             


Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser Val Lys  Val Leu Val 
    1160                 1165                 1170             


Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile 
    1175                 1180                 1185             


Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu 
    1190                 1195                 1200             


Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1205                 1210                 1215             


Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn 
    1220                 1225                 1230             


Glu Leu  Val Leu Pro Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr 
    1235                 1240                 1245             


Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile 
    1250                 1255                 1260             


Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile 
    1265                 1270                 1275             


Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn 
    1280                 1285                 1290             


Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile 
    1295                 1300                 1305             


Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe 
    1310                 1315                 1320             


Gly Ala  Ser Gly Gly Phe Ser  Val Ser Met Val His  Ser Glu Arg 
    1325                 1330                 1335             


Leu Pro  Leu Leu Met Asn Arg  Asn Gly Ile Arg Phe  Gln Ser Glu 
    1340                 1345                 1350             


Val Ile  Asp Pro Asn Pro Pro  Lys Lys Val Lys Trp  Val Leu Thr 
    1355                 1360                 1365             


Gly Ser  Gly Gly Asn Met Phe  Thr Ala Asn Val Arg  Gly Ser Asn 
    1370                 1375                 1380             


Val Ser  Ser Leu Val Lys His  Thr Trp Ser Glu Ala  Phe Val Ser 
    1385                 1390                 1395             


Leu Ile  Glu Asp Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1400                 1405                 1410             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1415                 1420                 1425             


Asp Leu  Ser Gln Leu Gly Gly  Asp 
    1430                 1435     


<210>  45
<211>  1222
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI

<400>  45

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Arg Ile Asn Lys Ile Arg Lys 
225                 230                 235                 240 


Lys Leu Ser Ala Asp Asn Ala Thr Lys Pro Val Ser Arg Ser Gly Pro 
                245                 250                 255     


Met Lys Thr Leu Leu Val Arg Val Met Thr Asp Asp Leu Lys Lys Arg 
            260                 265                 270         


Leu Glu Lys Arg Arg Lys Lys Pro Glu Val Met Pro Gln Val Ile Ser 
        275                 280                 285             


Asn Asn Ala Ala Asn Asn Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys 
    290                 295                 300                 


Met Lys Glu Ala Ile Leu Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp 
305                 310                 315                 320 


His Val Gly Leu Met Cys Lys Phe Ala Gln Pro Ala Ser Lys Lys Ile 
                325                 330                 335     


Asp Gln Asn Lys Leu Lys Pro Glu Met Asp Glu Lys Gly Asn Leu Thr 
            340                 345                 350         


Thr Ala Gly Phe Ala Cys Ser Gln Cys Gly Gln Pro Leu Phe Val Tyr 
        355                 360                 365             


Lys Leu Glu Gln Val Ser Glu Lys Gly Lys Ala Tyr Thr Asn Tyr Phe 
    370                 375                 380                 


Gly Arg Cys Asn Val Ala Glu His Glu Lys Leu Ile Leu Leu Ala Gln 
385                 390                 395                 400 


Leu Lys Pro Glu Lys Asp Ser Asp Glu Ala Val Thr Tyr Ser Leu Gly 
                405                 410                 415     


Ala Ser Lys Phe Gly Gln Arg Ala Leu Asp Phe Tyr Ser Ile His Val 
            420                 425                 430         


Thr Lys Glu Ser Thr His Pro Val Lys Pro Leu Ala Gln Ile Ala Gly 
        435                 440                 445             


Asn Arg Tyr Ala Ser Gly Pro Val Gly Lys Ala Leu Ser Asp Ala Cys 
    450                 455                 460                 


Met Gly Thr Ile Ala Ser Phe Leu Ser Lys Tyr Gln Asp Ile Ile Ile 
465                 470                 475                 480 


Glu His Gln Lys Val Val Lys Gly Asn Gln Lys Arg Leu Glu Ser Leu 
                485                 490                 495     


Arg Glu Leu Ala Gly Lys Glu Asn Leu Glu Tyr Pro Ser Val Thr Leu 
            500                 505                 510         


Pro Pro Gln Pro His Thr Lys Glu Gly Val Asp Ala Tyr Asn Glu Val 
        515                 520                 525             


Ile Ala Arg Val Arg Met Trp Val Asn Leu Asn Leu Trp Gln Lys Leu 
    530                 535                 540                 


Lys Leu Ser Arg Asp Asp Ala Lys Pro Leu Leu Arg Leu Lys Gly Phe 
545                 550                 555                 560 


Pro Ser Phe Pro Val Val Glu Arg Arg Glu Asn Glu Val Asp Trp Trp 
                565                 570                 575     


Asn Thr Ile Asn Glu Val Lys Lys Leu Ile Asp Ala Lys Arg Asp Met 
            580                 585                 590         


Gly Arg Val Phe Trp Ser Gly Val Thr Ala Glu Lys Arg Asn Thr Ile 
        595                 600                 605             


Leu Glu Gly Tyr Asn Tyr Leu Pro Asn Glu Asn Asp His Lys Lys Arg 
    610                 615                 620                 


Glu Gly Ser Leu Glu Asn Pro Lys Lys Pro Ala Lys Arg Gln Phe Gly 
625                 630                 635                 640 


Asp Leu Leu Leu Tyr Leu Glu Lys Lys Tyr Ala Gly Asp Trp Gly Lys 
                645                 650                 655     


Val Phe Asp Glu Ala Trp Glu Arg Ile Asp Lys Lys Ile Ala Gly Leu 
            660                 665                 670         


Thr Ser His Ile Glu Arg Glu Glu Ala Arg Asn Ala Glu Asp Ala Gln 
        675                 680                 685             


Ser Lys Ala Val Leu Thr Asp Trp Leu Arg Ala Lys Ala Ser Phe Val 
    690                 695                 700                 


Leu Glu Arg Leu Lys Glu Met Asp Glu Lys Glu Phe Tyr Ala Cys Glu 
705                 710                 715                 720 


Ile Gln Leu Gln Lys Trp Tyr Gly Asp Leu Arg Gly Asn Pro Phe Ala 
                725                 730                 735     


Val Glu Ala Glu Asn Arg Val Val Asp Ile Ser Ala Phe Ser Ile Gly 
            740                 745                 750         


Ser Asp Gly His Ser Ile Gln Tyr Arg Asn Leu Leu Ala Trp Lys Tyr 
        755                 760                 765             


Leu Glu Asn Gly Lys Arg Glu Phe Tyr Leu Leu Met Asn Tyr Gly Lys 
    770                 775                 780                 


Lys Gly Arg Ile Arg Phe Thr Asp Gly Thr Asp Ile Lys Lys Ser Gly 
785                 790                 795                 800 


Lys Trp Gln Gly Leu Leu Tyr Gly Gly Gly Lys Ala Lys Val Ile Asp 
                805                 810                 815     


Leu Thr Phe Asp Pro Asp Asp Glu Gln Leu Ile Ile Leu Pro Leu Ala 
            820                 825                 830         


Phe Gly Thr Arg Gln Gly Arg Glu Phe Ile Trp Asn Asp Leu Leu Ser 
        835                 840                 845             


Leu Glu Thr Gly Leu Ile Lys Leu Ala Asn Gly Arg Val Ile Glu Lys 
    850                 855                 860                 


Thr Ile Tyr Asn Lys Lys Ile Gly Arg Asp Glu Pro Ala Leu Phe Val 
865                 870                 875                 880 


Ala Leu Thr Phe Glu Arg Arg Glu Val Val Asp Pro Ser Asn Ile Lys 
                885                 890                 895     


Pro Val Asn Leu Ile Gly Val Ala Arg Gly Glu Asn Ile Pro Ala Val 
            900                 905                 910         


Ile Ala Thr Leu Asp Pro Glu Gly Cys Pro Leu Pro Glu Phe Lys Asp 
        915                 920                 925             


Ser Ser Gly Gly Pro Thr Asp Ile Leu Arg Ile Gly Glu Gly Tyr Lys 
    930                 935                 940                 


Glu Lys Gln Arg Ala Ile Gln Ala Ala Lys Glu Val Glu Gln Arg Arg 
945                 950                 955                 960 


Ala Gly Gly Tyr Ser Arg Lys Phe Ala Ser Lys Ser Arg Asn Leu Ala 
                965                 970                 975     


Asp Asp Met Val Arg Asn Ser Ala Arg Asp Leu Phe Tyr His Ala Val 
            980                 985                 990         


Thr His Asp Ala Val Leu Val Phe  Ala Asn Leu Ser Arg  Gly Phe Gly 
        995                 1000                 1005             


Arg Gln  Gly Lys Arg Thr Phe  Met Thr Glu Arg Gln  Tyr Thr Lys 
    1010                 1015                 1020             


Met Glu  Asp Trp Leu Thr Ala  Lys Leu Ala Tyr Glu  Gly Leu Thr 
    1025                 1030                 1035             


Ser Lys  Thr Tyr Leu Ser Lys  Thr Leu Ala Gln Tyr  Thr Ser Lys 
    1040                 1045                 1050             


Thr Cys  Ser Asn Cys Gly Phe  Thr Ile Thr Thr Ala  Asp Tyr Asp 
    1055                 1060                 1065             


Gly Met  Leu Val Arg Leu Lys  Lys Thr Ser Asp Gly  Trp Ala Thr 
    1070                 1075                 1080             


Thr Leu  Asn Asn Lys Glu Leu  Lys Ala Glu Gly Gln  Ile Thr Tyr 
    1085                 1090                 1095             


Tyr Asn  Arg Tyr Lys Arg Gln  Thr Val Glu Lys Glu  Leu Ser Ala 
    1100                 1105                 1110             


Glu Leu  Asp Arg Leu Ser Glu  Glu Ser Gly Asn Asn  Asp Ile Ser 
    1115                 1120                 1125             


Lys Trp  Thr Lys Gly Arg Arg  Asp Glu Ala Leu Phe  Leu Leu Lys 
    1130                 1135                 1140             


Lys Arg  Phe Ser His Gly Ser  Arg Pro Val Gln Glu  Gln Phe Val 
    1145                 1150                 1155             


Cys Leu  Asp Cys Gly His Glu  Val His Ala Ala Glu  Gln Ala Ala 
    1160                 1165                 1170             


Leu Asn  Ile Ala Arg Ser Trp  Leu Phe Leu Asn Ser  Asn Ser Thr 
    1175                 1180                 1185             


Glu Phe  Lys Ser Tyr Lys Ser  Gly Lys Gln Pro Phe  Val Gly Ala 
    1190                 1195                 1200             


Trp Gln  Ala Phe Tyr Lys Arg  Arg Leu Lys Glu Val  Trp Lys Pro 
    1205                 1210                 1215             


Asn Ala  Gly Ala 
    1220         


<210>  46
<211>  1135
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI Dntsb

<400>  46

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Arg Ile Asn Lys Ile Arg Lys 
225                 230                 235                 240 


Lys Leu Ser Ala Asp Asn Ala Thr Lys Pro Val Ser Arg Ser Gly Pro 
                245                 250                 255     


Met Lys Thr Leu Leu Val Arg Val Met Thr Asp Asp Leu Lys Lys Arg 
            260                 265                 270         


Leu Glu Lys Arg Arg Lys Lys Pro Glu Val Met Pro Gln Val Ile Ser 
        275                 280                 285             


Asn Asn Ala Ala Asn Asn Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys 
    290                 295                 300                 


Met Lys Glu Ala Ile Leu Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp 
305                 310                 315                 320 


His Val Gly Leu Met Cys Lys Phe Ala Ala Ser Lys Phe Gly Gln Arg 
                325                 330                 335     


Ala Leu Asp Phe Tyr Ser Ile His Val Thr Lys Glu Ser Thr His Pro 
            340                 345                 350         


Val Lys Pro Leu Ala Gln Ile Ala Gly Asn Arg Tyr Ala Ser Gly Pro 
        355                 360                 365             


Val Gly Lys Ala Leu Ser Asp Ala Cys Met Gly Thr Ile Ala Ser Phe 
    370                 375                 380                 


Leu Ser Lys Tyr Gln Asp Ile Ile Ile Glu His Gln Lys Val Val Lys 
385                 390                 395                 400 


Gly Asn Gln Lys Arg Leu Glu Ser Leu Arg Glu Leu Ala Gly Lys Glu 
                405                 410                 415     


Asn Leu Glu Tyr Pro Ser Val Thr Leu Pro Pro Gln Pro His Thr Lys 
            420                 425                 430         


Glu Gly Val Asp Ala Tyr Asn Glu Val Ile Ala Arg Val Arg Met Trp 
        435                 440                 445             


Val Asn Leu Asn Leu Trp Gln Lys Leu Lys Leu Ser Arg Asp Asp Ala 
    450                 455                 460                 


Lys Pro Leu Leu Arg Leu Lys Gly Phe Pro Ser Phe Pro Val Val Glu 
465                 470                 475                 480 


Arg Arg Glu Asn Glu Val Asp Trp Trp Asn Thr Ile Asn Glu Val Lys 
                485                 490                 495     


Lys Leu Ile Asp Ala Lys Arg Asp Met Gly Arg Val Phe Trp Ser Gly 
            500                 505                 510         


Val Thr Ala Glu Lys Arg Asn Thr Ile Leu Glu Gly Tyr Asn Tyr Leu 
        515                 520                 525             


Pro Asn Glu Asn Asp His Lys Lys Arg Glu Gly Ser Leu Glu Asn Pro 
    530                 535                 540                 


Lys Lys Pro Ala Lys Arg Gln Phe Gly Asp Leu Leu Leu Tyr Leu Glu 
545                 550                 555                 560 


Lys Lys Tyr Ala Gly Asp Trp Gly Lys Val Phe Asp Glu Ala Trp Glu 
                565                 570                 575     


Arg Ile Asp Lys Lys Ile Ala Gly Leu Thr Ser His Ile Glu Arg Glu 
            580                 585                 590         


Glu Ala Arg Asn Ala Glu Asp Ala Gln Ser Lys Ala Val Leu Thr Asp 
        595                 600                 605             


Trp Leu Arg Ala Lys Ala Ser Phe Val Leu Glu Arg Leu Lys Glu Met 
    610                 615                 620                 


Asp Glu Lys Glu Phe Tyr Ala Cys Glu Ile Gln Leu Gln Lys Trp Tyr 
625                 630                 635                 640 


Gly Asp Leu Arg Gly Asn Pro Phe Ala Val Glu Ala Glu Asn Arg Val 
                645                 650                 655     


Val Asp Ile Ser Ala Phe Ser Ile Gly Ser Asp Gly His Ser Ile Gln 
            660                 665                 670         


Tyr Arg Asn Leu Leu Ala Trp Lys Tyr Leu Glu Asn Gly Lys Arg Glu 
        675                 680                 685             


Phe Tyr Leu Leu Met Asn Tyr Gly Lys Lys Gly Arg Ile Arg Phe Thr 
    690                 695                 700                 


Asp Gly Thr Asp Ile Lys Lys Ser Gly Lys Trp Gln Gly Leu Leu Tyr 
705                 710                 715                 720 


Gly Gly Gly Lys Ala Lys Val Ile Asp Leu Thr Phe Asp Pro Asp Asp 
                725                 730                 735     


Glu Gln Leu Ile Ile Leu Pro Leu Ala Phe Gly Thr Arg Gln Gly Arg 
            740                 745                 750         


Glu Phe Ile Trp Asn Asp Leu Leu Ser Leu Glu Thr Gly Leu Ile Lys 
        755                 760                 765             


Leu Ala Asn Gly Arg Val Ile Glu Lys Thr Ile Tyr Asn Lys Lys Ile 
    770                 775                 780                 


Gly Arg Asp Glu Pro Ala Leu Phe Val Ala Leu Thr Phe Glu Arg Arg 
785                 790                 795                 800 


Glu Val Val Asp Pro Ser Asn Ile Lys Pro Val Asn Leu Ile Gly Val 
                805                 810                 815     


Ala Arg Gly Glu Asn Ile Pro Ala Val Ile Ala Thr Leu Asp Pro Glu 
            820                 825                 830         


Gly Cys Pro Leu Pro Glu Phe Lys Asp Ser Ser Gly Gly Pro Thr Asp 
        835                 840                 845             


Ile Leu Arg Ile Gly Glu Gly Tyr Lys Glu Lys Gln Arg Ala Ile Gln 
    850                 855                 860                 


Ala Ala Lys Glu Val Glu Gln Arg Arg Ala Gly Gly Tyr Ser Arg Lys 
865                 870                 875                 880 


Phe Ala Ser Lys Ser Arg Asn Leu Ala Asp Asp Met Val Arg Asn Ser 
                885                 890                 895     


Ala Arg Asp Leu Phe Tyr His Ala Val Thr His Asp Ala Val Leu Val 
            900                 905                 910         


Phe Ala Asn Leu Ser Arg Gly Phe Gly Arg Gln Gly Lys Arg Thr Phe 
        915                 920                 925             


Met Thr Glu Arg Gln Tyr Thr Lys Met Glu Asp Trp Leu Thr Ala Lys 
    930                 935                 940                 


Leu Ala Tyr Glu Gly Leu Thr Ser Lys Thr Tyr Leu Ser Lys Thr Leu 
945                 950                 955                 960 


Ala Gln Tyr Thr Ser Lys Thr Cys Ser Asn Cys Gly Phe Thr Ile Thr 
                965                 970                 975     


Thr Ala Asp Tyr Asp Gly Met Leu Val Arg Leu Lys Lys Thr Ser Asp 
            980                 985                 990         


Gly Trp Ala Thr Thr Leu Asn Asn  Lys Glu Leu Lys Ala  Glu Gly Gln 
        995                 1000                 1005             


Ile Thr  Tyr Tyr Asn Arg Tyr  Lys Arg Gln Thr Val  Glu Lys Glu 
    1010                 1015                 1020             


Leu Ser  Ala Glu Leu Asp Arg  Leu Ser Glu Glu Ser  Gly Asn Asn 
    1025                 1030                 1035             


Asp Ile  Ser Lys Trp Thr Lys  Gly Arg Arg Asp Glu  Ala Leu Phe 
    1040                 1045                 1050             


Leu Leu  Lys Lys Arg Phe Ser  His Gly Ser Arg Pro  Val Gln Glu 
    1055                 1060                 1065             


Gln Phe  Val Cys Leu Asp Cys  Gly His Glu Val His  Ala Ala Glu 
    1070                 1075                 1080             


Gln Ala  Ala Leu Asn Ile Ala  Arg Ser Trp Leu Phe  Leu Asn Ser 
    1085                 1090                 1095             


Asn Ser  Thr Glu Phe Lys Ser  Tyr Lys Ser Gly Lys  Gln Pro Phe 
    1100                 1105                 1110             


Val Gly  Ala Trp Gln Ala Phe  Tyr Lys Arg Arg Leu  Lys Glu Val 
    1115                 1120                 1125             


Trp Lys  Pro Asn Ala Gly Ala  
    1130                 1135 


<210>  47
<211>  1132
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI dTSL

<400>  47

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Arg Ile Asn Lys Ile Arg Lys 
225                 230                 235                 240 


Lys Leu Ser Ala Asp Asn Ala Thr Lys Pro Val Ser Arg Ser Gly Pro 
                245                 250                 255     


Met Lys Thr Leu Leu Val Arg Val Met Thr Asp Asp Leu Lys Lys Arg 
            260                 265                 270         


Leu Glu Lys Arg Arg Lys Lys Pro Glu Val Met Pro Gln Val Ile Ser 
        275                 280                 285             


Asn Asn Ala Ala Asn Asn Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys 
    290                 295                 300                 


Met Lys Glu Ala Ile Leu Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp 
305                 310                 315                 320 


His Val Gly Leu Met Cys Lys Phe Ala Gln Pro Ala Ser Lys Lys Ile 
                325                 330                 335     


Asp Gln Asn Lys Leu Lys Pro Glu Met Asp Glu Lys Gly Asn Leu Thr 
            340                 345                 350         


Thr Ala Gly Phe Ala Cys Ser Gln Cys Gly Gln Pro Leu Phe Val Tyr 
        355                 360                 365             


Lys Leu Glu Gln Val Ser Glu Lys Gly Lys Ala Tyr Thr Asn Tyr Phe 
    370                 375                 380                 


Gly Arg Cys Asn Val Ala Glu His Glu Lys Leu Ile Leu Leu Ala Gln 
385                 390                 395                 400 


Leu Lys Pro Glu Lys Asp Ser Asp Glu Ala Val Thr Tyr Ser Leu Gly 
                405                 410                 415     


Ala Ser Lys Phe Gly Gln Arg Ala Leu Asp Phe Tyr Ser Ile His Val 
            420                 425                 430         


Thr Lys Glu Ser Thr His Pro Val Lys Pro Leu Ala Gln Ile Ala Gly 
        435                 440                 445             


Asn Arg Tyr Ala Ser Gly Pro Val Gly Lys Ala Leu Ser Asp Ala Cys 
    450                 455                 460                 


Met Gly Thr Ile Ala Ser Phe Leu Ser Lys Tyr Gln Asp Ile Ile Ile 
465                 470                 475                 480 


Glu His Gln Lys Val Val Lys Gly Asn Gln Lys Arg Leu Glu Ser Leu 
                485                 490                 495     


Arg Glu Leu Ala Gly Lys Glu Asn Leu Glu Tyr Pro Ser Val Thr Leu 
            500                 505                 510         


Pro Pro Gln Pro His Thr Lys Glu Gly Val Asp Ala Tyr Asn Glu Val 
        515                 520                 525             


Ile Ala Arg Val Arg Met Trp Val Asn Leu Asn Leu Trp Gln Lys Leu 
    530                 535                 540                 


Lys Leu Ser Arg Asp Asp Ala Lys Pro Leu Leu Arg Leu Lys Gly Phe 
545                 550                 555                 560 


Pro Ser Phe Pro Val Val Glu Arg Arg Glu Asn Glu Val Asp Trp Trp 
                565                 570                 575     


Asn Thr Ile Asn Glu Val Lys Lys Leu Ile Asp Ala Lys Arg Asp Met 
            580                 585                 590         


Gly Arg Val Phe Trp Ser Gly Val Thr Ala Glu Lys Arg Asn Thr Ile 
        595                 600                 605             


Leu Glu Gly Tyr Asn Tyr Leu Pro Asn Glu Asn Asp His Lys Lys Arg 
    610                 615                 620                 


Glu Gly Ser Leu Glu Asn Pro Lys Lys Pro Ala Lys Arg Gln Phe Gly 
625                 630                 635                 640 


Asp Leu Leu Leu Tyr Leu Glu Lys Lys Tyr Ala Gly Asp Trp Gly Lys 
                645                 650                 655     


Val Phe Asp Glu Ala Trp Glu Arg Ile Asp Lys Lys Ile Ala Gly Leu 
            660                 665                 670         


Thr Ser His Ile Glu Arg Glu Glu Ala Arg Asn Ala Glu Asp Ala Gln 
        675                 680                 685             


Ser Lys Ala Val Leu Thr Asp Trp Leu Arg Ala Lys Ala Ser Phe Val 
    690                 695                 700                 


Leu Glu Arg Leu Lys Glu Met Asp Glu Lys Glu Phe Tyr Ala Cys Glu 
705                 710                 715                 720 


Ile Gln Leu Gln Lys Trp Tyr Gly Asp Leu Arg Gly Asn Pro Phe Ala 
                725                 730                 735     


Val Glu Ala Glu Asn Arg Val Val Asp Ile Ser Ala Phe Ser Ile Gly 
            740                 745                 750         


Ser Asp Gly His Ser Ile Gln Tyr Arg Asn Leu Leu Ala Trp Lys Tyr 
        755                 760                 765             


Leu Glu Asn Gly Lys Arg Glu Phe Tyr Leu Leu Met Asn Tyr Gly Lys 
    770                 775                 780                 


Lys Gly Arg Ile Arg Phe Thr Asp Gly Thr Asp Ile Lys Lys Ser Gly 
785                 790                 795                 800 


Lys Trp Gln Gly Leu Leu Tyr Gly Gly Gly Lys Ala Lys Val Ile Asp 
                805                 810                 815     


Leu Thr Phe Asp Pro Asp Asp Glu Gln Leu Ile Ile Leu Pro Leu Ala 
            820                 825                 830         


Phe Gly Thr Arg Gln Gly Arg Glu Phe Ile Trp Asn Asp Leu Leu Ser 
        835                 840                 845             


Leu Glu Thr Gly Leu Ile Lys Leu Ala Asn Gly Arg Val Ile Glu Lys 
    850                 855                 860                 


Thr Ile Tyr Asn Lys Lys Ile Gly Arg Asp Glu Pro Ala Leu Phe Val 
865                 870                 875                 880 


Ala Leu Thr Phe Glu Arg Arg Glu Val Val Asp Pro Ser Asn Ile Lys 
                885                 890                 895     


Pro Val Asn Leu Ile Gly Val Ala Arg Gly Glu Asn Ile Pro Ala Val 
            900                 905                 910         


Ile Ala Thr Leu Asp Pro Glu Gly Cys Pro Leu Pro Glu Phe Lys Asp 
        915                 920                 925             


Ser Ser Gly Gly Pro Thr Asp Ile Leu Arg Ile Gly Glu Gly Tyr Lys 
    930                 935                 940                 


Glu Lys Gln Arg Ala Ile Gln Ala Ala Lys Glu Val Glu Gln Arg Arg 
945                 950                 955                 960 


Ala Gly Gly Tyr Ser Arg Lys Phe Ala Ser Lys Ser Arg Asn Leu Ala 
                965                 970                 975     


Asp Asp Met Val Arg Asn Ser Ala Arg Asp Leu Phe Tyr His Ala Val 
            980                 985                 990         


Thr His Asp Ala Val Leu Val Phe  Ala Asn Leu Ser Arg  Gly Phe Gly 
        995                 1000                 1005             


Arg Gln  Gly Lys Arg Thr Phe  Met Thr Glu Arg Gln  Tyr Thr Lys 
    1010                 1015                 1020             


Met Glu  Asp Trp Leu Thr Ala  Lys Leu Ala Tyr Glu  Gly Leu Thr 
    1025                 1030                 1035             


Ser Lys  Thr Tyr Leu Ser Lys  Thr Leu Ala Gln Tyr  Thr Ser Lys 
    1040                 1045                 1050             


Thr Cys  Ser Asn Cys Gly Ser  Arg Pro Val Gln Glu  Gln Phe Val 
    1055                 1060                 1065             


Cys Leu  Asp Cys Gly His Glu  Val His Ala Ala Glu  Gln Ala Ala 
    1070                 1075                 1080             


Leu Asn  Ile Ala Arg Ser Trp  Leu Phe Leu Asn Ser  Asn Ser Thr 
    1085                 1090                 1095             


Glu Phe  Lys Ser Tyr Lys Ser  Gly Lys Gln Pro Phe  Val Gly Ala 
    1100                 1105                 1110             


Trp Gln  Ala Phe Tyr Lys Arg  Arg Leu Lys Glu Val  Trp Lys Pro 
    1115                 1120                 1125             


Asn Ala  Gly Ala 
    1130         


<210>  48
<211>  1045
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI dNTSB dTSL

<400>  48

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Arg Ile Asn Lys Ile Arg Lys 
225                 230                 235                 240 


Lys Leu Ser Ala Asp Asn Ala Thr Lys Pro Val Ser Arg Ser Gly Pro 
                245                 250                 255     


Met Lys Thr Leu Leu Val Arg Val Met Thr Asp Asp Leu Lys Lys Arg 
            260                 265                 270         


Leu Glu Lys Arg Arg Lys Lys Pro Glu Val Met Pro Gln Val Ile Ser 
        275                 280                 285             


Asn Asn Ala Ala Asn Asn Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys 
    290                 295                 300                 


Met Lys Glu Ala Ile Leu Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp 
305                 310                 315                 320 


His Val Gly Leu Met Cys Lys Phe Ala Ala Ser Lys Phe Gly Gln Arg 
                325                 330                 335     


Ala Leu Asp Phe Tyr Ser Ile His Val Thr Lys Glu Ser Thr His Pro 
            340                 345                 350         


Val Lys Pro Leu Ala Gln Ile Ala Gly Asn Arg Tyr Ala Ser Gly Pro 
        355                 360                 365             


Val Gly Lys Ala Leu Ser Asp Ala Cys Met Gly Thr Ile Ala Ser Phe 
    370                 375                 380                 


Leu Ser Lys Tyr Gln Asp Ile Ile Ile Glu His Gln Lys Val Val Lys 
385                 390                 395                 400 


Gly Asn Gln Lys Arg Leu Glu Ser Leu Arg Glu Leu Ala Gly Lys Glu 
                405                 410                 415     


Asn Leu Glu Tyr Pro Ser Val Thr Leu Pro Pro Gln Pro His Thr Lys 
            420                 425                 430         


Glu Gly Val Asp Ala Tyr Asn Glu Val Ile Ala Arg Val Arg Met Trp 
        435                 440                 445             


Val Asn Leu Asn Leu Trp Gln Lys Leu Lys Leu Ser Arg Asp Asp Ala 
    450                 455                 460                 


Lys Pro Leu Leu Arg Leu Lys Gly Phe Pro Ser Phe Pro Val Val Glu 
465                 470                 475                 480 


Arg Arg Glu Asn Glu Val Asp Trp Trp Asn Thr Ile Asn Glu Val Lys 
                485                 490                 495     


Lys Leu Ile Asp Ala Lys Arg Asp Met Gly Arg Val Phe Trp Ser Gly 
            500                 505                 510         


Val Thr Ala Glu Lys Arg Asn Thr Ile Leu Glu Gly Tyr Asn Tyr Leu 
        515                 520                 525             


Pro Asn Glu Asn Asp His Lys Lys Arg Glu Gly Ser Leu Glu Asn Pro 
    530                 535                 540                 


Lys Lys Pro Ala Lys Arg Gln Phe Gly Asp Leu Leu Leu Tyr Leu Glu 
545                 550                 555                 560 


Lys Lys Tyr Ala Gly Asp Trp Gly Lys Val Phe Asp Glu Ala Trp Glu 
                565                 570                 575     


Arg Ile Asp Lys Lys Ile Ala Gly Leu Thr Ser His Ile Glu Arg Glu 
            580                 585                 590         


Glu Ala Arg Asn Ala Glu Asp Ala Gln Ser Lys Ala Val Leu Thr Asp 
        595                 600                 605             


Trp Leu Arg Ala Lys Ala Ser Phe Val Leu Glu Arg Leu Lys Glu Met 
    610                 615                 620                 


Asp Glu Lys Glu Phe Tyr Ala Cys Glu Ile Gln Leu Gln Lys Trp Tyr 
625                 630                 635                 640 


Gly Asp Leu Arg Gly Asn Pro Phe Ala Val Glu Ala Glu Asn Arg Val 
                645                 650                 655     


Val Asp Ile Ser Ala Phe Ser Ile Gly Ser Asp Gly His Ser Ile Gln 
            660                 665                 670         


Tyr Arg Asn Leu Leu Ala Trp Lys Tyr Leu Glu Asn Gly Lys Arg Glu 
        675                 680                 685             


Phe Tyr Leu Leu Met Asn Tyr Gly Lys Lys Gly Arg Ile Arg Phe Thr 
    690                 695                 700                 


Asp Gly Thr Asp Ile Lys Lys Ser Gly Lys Trp Gln Gly Leu Leu Tyr 
705                 710                 715                 720 


Gly Gly Gly Lys Ala Lys Val Ile Asp Leu Thr Phe Asp Pro Asp Asp 
                725                 730                 735     


Glu Gln Leu Ile Ile Leu Pro Leu Ala Phe Gly Thr Arg Gln Gly Arg 
            740                 745                 750         


Glu Phe Ile Trp Asn Asp Leu Leu Ser Leu Glu Thr Gly Leu Ile Lys 
        755                 760                 765             


Leu Ala Asn Gly Arg Val Ile Glu Lys Thr Ile Tyr Asn Lys Lys Ile 
    770                 775                 780                 


Gly Arg Asp Glu Pro Ala Leu Phe Val Ala Leu Thr Phe Glu Arg Arg 
785                 790                 795                 800 


Glu Val Val Asp Pro Ser Asn Ile Lys Pro Val Asn Leu Ile Gly Val 
                805                 810                 815     


Ala Arg Gly Glu Asn Ile Pro Ala Val Ile Ala Thr Leu Asp Pro Glu 
            820                 825                 830         


Gly Cys Pro Leu Pro Glu Phe Lys Asp Ser Ser Gly Gly Pro Thr Asp 
        835                 840                 845             


Ile Leu Arg Ile Gly Glu Gly Tyr Lys Glu Lys Gln Arg Ala Ile Gln 
    850                 855                 860                 


Ala Ala Lys Glu Val Glu Gln Arg Arg Ala Gly Gly Tyr Ser Arg Lys 
865                 870                 875                 880 


Phe Ala Ser Lys Ser Arg Asn Leu Ala Asp Asp Met Val Arg Asn Ser 
                885                 890                 895     


Ala Arg Asp Leu Phe Tyr His Ala Val Thr His Asp Ala Val Leu Val 
            900                 905                 910         


Phe Ala Asn Leu Ser Arg Gly Phe Gly Arg Gln Gly Lys Arg Thr Phe 
        915                 920                 925             


Met Thr Glu Arg Gln Tyr Thr Lys Met Glu Asp Trp Leu Thr Ala Lys 
    930                 935                 940                 


Leu Ala Tyr Glu Gly Leu Thr Ser Lys Thr Tyr Leu Ser Lys Thr Leu 
945                 950                 955                 960 


Ala Gln Tyr Thr Ser Lys Thr Cys Ser Asn Cys Gly Ser Arg Pro Val 
                965                 970                 975     


Gln Glu Gln Phe Val Cys Leu Asp Cys Gly His Glu Val His Ala Ala 
            980                 985                 990         


Glu Gln Ala Ala Leu Asn Ile Ala  Arg Ser Trp Leu Phe  Leu Asn Ser 
        995                 1000                 1005             


Asn Ser  Thr Glu Phe Lys Ser  Tyr Lys Ser Gly Lys  Gln Pro Phe 
    1010                 1015                 1020             


Val Gly  Ala Trp Gln Ala Phe  Tyr Lys Arg Arg Leu  Lys Glu Val 
    1025                 1030                 1035             


Trp Lys  Pro Asn Ala Gly Ala  
    1040                 1045 


<210>  49
<211>  15
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Target site on PDS exon 14

<400>  49
tccagatgaa agtgc                                                        15


<210>  50
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  PDS exon 14 Right target sgRNA

<400>  50
cuccaaaaac ccguuuugau guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  51
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  PDS exon 14 Left target sgRNA

<400>  51
gguugugaac uaaugggaca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  52
<211>  76
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  NekP-AtU61P cassette- Right Cas9 sgRNA scaffold

<400>  52
gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt       60

ggcaccgagt cggtgc                                                       76


<210>  53
<211>  76
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  NekP-AtU61P cassette- Left Cas9 sgRNA scaffold

<400>  53
gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta       60

tttctagctc taaaac                                                       76


<210>  54
<211>  644
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  NekP-AtU61P cassette


<220>
<221>  misc_feature
<222>  (112)..(131)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (515)..(534)
<223>  n is a, c, g, or t

<400>  54
ggatccattt aaattctaga ggcgcgccaa aaaaagcacc gactcggtgc cactttttca       60

agttgataac ggactagcct tattttaact tgctatttct agctctaaaa cnnnnnnnnn      120

nnnnnnnnnn naatcgctat gtcgactcta tcattatata aactaagctg ctatatatca      180

cctgatcgat gtgggacttt tgatcactcc agaaatctca aaattccggc agaacaattt      240

tgaatctcga tccgtagaaa cgagacggtc attgttttag ttccaccacg attatatttg      300

aaatttacgt gagtgtgagt gagacttgca taagaaaata aaatctttag ttgggaaaaa      360

attcaataat ataaatgggc ttgagaagga agcgagggat aggccttttt ctaaaatagg      420

cccatttaag ctattaacaa tcttcaaaag taccacagcg cttaggtaaa gaaagcagct      480

gagtttatat atggttagag acgaagtagt gattnnnnnn nnnnnnnnnn nnnngtttta      540

gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc      600

gagtcggtgc ttttttttaa ttaataggga taacagggta atta                       644


<210>  55
<211>  644
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  NekP-AtU61P containing PDS specific sgRNA

<400>  55
ggatccattt aaattctaga ggcgcgccaa aaaaagcacc gactcggtgc cactttttca       60

agttgataac ggactagcct tattttaact tgctatttct agctctaaaa ctgtcccatt      120

agttcacaac caatcgctat gtcgactcta tcattatata aactaagctg ctatatatca      180

cctgatcgat gtgggacttt tgatcactcc agaaatctca aaattccggc agaacaattt      240

tgaatctcga tccgtagaaa cgagacggtc attgttttag ttccaccacg attatatttg      300

aaatttacgt gagtgtgagt gagacttgca taagaaaata aaatctttag ttgggaaaaa      360

attcaataat ataaatgggc ttgagaagga agcgagggat aggccttttt ctaaaatagg      420

cccatttaag ctattaacaa tcttcaaaag taccacagcg cttaggtaaa gaaagcagct      480

gagtttatat atggttagag acgaagtagt gattctccaa aaacccgttt tgatgtttta      540

gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc      600

gagtcggtgc ttttttttaa ttaataggga taacagggta atta                       644


<210>  56
<211>  1617
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, nucleoplasmin NLS

<400>  56

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
                805                 810                 815     


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
            820                 825                 830         


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
        835                 840                 845             


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
    850                 855                 860                 


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
865                 870                 875                 880 


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
                885                 890                 895     


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
            900                 905                 910         


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
        915                 920                 925             


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
    930                 935                 940                 


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
945                 950                 955                 960 


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
                965                 970                 975     


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
            980                 985                 990         


Asn Ile Val Ile Glu Met Ala Arg  Glu Asn Gln Thr Thr  Thr Lys Gly 
        995                 1000                 1005             


Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg Ile Glu  Glu Gly Ile 
    1010                 1015                 1020             


Lys Glu  Leu Glu Ser Gln Ile  Leu Lys Glu Asn Pro  Val Glu Asn 
    1025                 1030                 1035             


Thr Gln  Leu Gln Asn Glu Lys  Leu Tyr Leu Tyr Tyr  Leu Gln Asn 
    1040                 1045                 1050             


Gly Arg  Asp Met Tyr Val Asp  Gln Glu Leu Asp Ile  Asn Arg Leu 
    1055                 1060                 1065             


Ser Asp  Tyr Asp Val Asp Ala  Ile Val Pro Gln Ser  Phe Ile Lys 
    1070                 1075                 1080             


Asp Asp  Ser Ile Asp Asn Lys  Val Leu Thr Arg Ser  Val Glu Asn 
    1085                 1090                 1095             


Arg Gly  Lys Ser Asp Asn Val  Pro Ser Glu Glu Val  Val Lys Lys 
    1100                 1105                 1110             


Met Lys  Asn Tyr Trp Arg Gln  Leu Leu Asn Ala Lys  Leu Ile Thr 
    1115                 1120                 1125             


Gln Arg  Lys Phe Asp Asn Leu  Thr Lys Ala Glu Arg  Gly Gly Leu 
    1130                 1135                 1140             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1145                 1150                 1155             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1160                 1165                 1170             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1175                 1180                 1185             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1190                 1195                 1200             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1205                 1210                 1215             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1220                 1225                 1230             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1235                 1240                 1245             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1250                 1255                 1260             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1265                 1270                 1275             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1280                 1285                 1290             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1295                 1300                 1305             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1310                 1315                 1320             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1325                 1330                 1335             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1340                 1345                 1350             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1355                 1360                 1365             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1370                 1375                 1380             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1385                 1390                 1395             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1400                 1405                 1410             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1415                 1420                 1425             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1430                 1435                 1440             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1445                 1450                 1455             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1460                 1465                 1470             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1475                 1480                 1485             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1490                 1495                 1500             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1505                 1510                 1515             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1520                 1525                 1530             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1535                 1540                 1545             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1550                 1555                 1560             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1565                 1570                 1575             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1580                 1585                 1590             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1595                 1600                 1605             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1610                 1615         


<210>  57
<211>  905
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  2X35SP from pSAT6

<400>  57
caacatgtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa       60

gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc      120

cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac      180

aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt      240

cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg      300

tcttcaaagc aagtggattg atgtgataac atggtggagc acgacacact tgtctactcc      360

aaaaatatca aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg      420

gtaatatccg gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag      480

atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc      540

gttgaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc      600

gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc      660

actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa      720

ggaagttcat ttcatttgga gaggacgtcg agagttctca acacaacata tacaaaacaa      780

acgaatctca agcaatcaag cattctactt ctattgcagc aatttaaatc atttctttta      840

aagcaaaagc aattttctga aaattttcac catttacgaa cgatagtccg gactcagatc      900

tcgag                                                                  905


<210>  58
<211>  4806
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Plant optimized dScCAS9-FokI

<400>  58
atggcaccta agaagaagcg aaaggttggg ggatctggga gtcaactagt taagtcggag       60

cttgaggaga aaaagtctga gctcaggcat aaattaaagt acgttcccca tgaatacatt      120

gagcttattg aaatcgctag aaattctaca caagatcgta tcttggaaat gaaggtgatg      180

gagtttttca tgaaggttta cgggtacaga ggtaagcacc ttggaggatc gagaaagcca      240

gatggagcta tttacacagt aggtagtccc atagattatg gcgtaatcgt agataccaaa      300

gcttatagtg gcggttataa tttgcctatt ggtcaggccg atgaaatgca aaggtacgta      360

gaggagaatc agacacgaaa taagcatata aaccctaacg agtggtggaa agtttatcct      420

tcttcagtca cagaattcaa gttcttgttt gtcagtggac atttcaaagg taattataaa      480

gcgcaattaa caagactcaa tcatataacc aactgtaatg gtgcggtcct ttctgtggaa      540

gagctcttga ttggagggga gatgattaag gctggaactt taacgctaga ggaggttcgt      600

cgtaagttta ataacggaga aattaatttt tccggatctg aaacaccggg aaccagtgag      660

tccgctactc ctgagtctat ggagaagaag tactccattg gattggctat tggaactaac      720

tccgtgggtt gggctgtgat tacggatgat tataaggttc cgagcaaaaa atttaaggta      780

ctcggcaaca ccaacaggaa atcgataaaa aagaatttga tgggtgctct tttattcgat      840

agcggcgaga ctgccgaagc taccaggctc aaacgtacag caagacgacg ttatacaaga      900

agaaagaacc gtattagata cttgcaggag attttcgcaa acgagatggc taagctcgat      960

gactcttttt ttcagcgttt ggaggagtcc tttcttgtcg aagaagataa aaaaaacgaa     1020

agacacccga tttttgggaa cctggctgat gaagttgcct accatcgaaa ttacccaaca     1080

atctaccatc ttagaaagaa gcttgcagat tctccggaaa aagccgatct tagactgatt     1140

tacctagctt tggcacacat tatcaaattc agagggcact tccttatcga gggtaagctt     1200

aatgcggaga attccgatgt tgctaagctt ttttatcagc taatccagac ttacaaccaa     1260

ctgttcgagg aaagcccact tgacgagatt gaggttgatg ctaaaggcat attgtcagct     1320

cgacttagta agtcgaagag attggagaag ctcatcgcgg ttttccctaa tgagaaaaaa     1380

aatggattgt ttgggaacat tatagcctta gctttgggat tgacgcctaa cttcaaaagc     1440

aacttcgatc tcactgaaga tgctaagctt caactttcaa aagacactta tgatgatgat     1500

ttagatgaac tcctcggaca aataggggat caatatgccg atttatttag cgctgcgaag     1560

aacctttcgg acgcaatctt gctctcagat atccttagat ctaattctga agtaacgaaa     1620

gctcctttat cagcgagcat ggtaaaaaga tatgatgagc atcatcaaga tttggcactt     1680

ttgaagacct tagtgcgaca acaattccca gagaaatatg ctgagatatt taaggatgac     1740

acaaagaacg ggtacgctgg atatgtcggc attggtatta agcatcgtaa gcgtacaacg     1800

aagttagcta cacaagagga gttttataag tttattaagc ctatattaga gaagatggac     1860

ggtgccgaag agttgctcgc taaactgaat cgagacgatt tgctacgaaa gcagagaact     1920

tttgataatg gaagcattcc tcaccaaatt cacctcaagg agcttcatgc aattttaagg     1980

agacaggagg agttttaccc atttcttaag gaaaaccgtg aaaaaataga gaaaatcttg     2040

acgttccgta ttccatacta tgttggaccg ttagcgagag gaaattctag attcgcctgg     2100

ctaacacgaa aatcagaaga ggcaataact ccatggaatt tcgaggaagt tgtggataag     2160

ggtgcaagcg cccaaagttt catcgaacgt atgactaact tcgatgaaca gttgccaaat     2220

aaaaaggtat tgcctaagca ttcgcttcta tacgagtact tcacagtgta taatgagcta     2280

actaaggtca aatatgtaac tgagcgaatg cgtaaaccag aattcctgag cggagaacaa     2340

aaaaaggcta ttgtagattt gctctttaag acgaacagaa aggttactgt taaacagctt     2400

aaggaagatt attttaagaa gattgaatgc tttgattctg ttgaaatcat aggagtggag     2460

gacaggttta atgcttcatt gggcacgtat catgaccttt tgaagattat caaggacaag     2520

gacttcttag acaacgagga aaacgaagat attttggagg acattgtctt gacattgaca     2580

ctgttcgaag acagggaaat gattgaagag aggttgaaaa catatgcgca tttgttcgat     2640

gataaggtta tgaagcagct gaagcgtcga cactataccg gctggggtag actgtctaga     2700

aaaatgataa atggtattcg tgataaacag agtgggaaga caattctaga ttttcttaaa     2760

agtgatgggt tttctaatcg aaattttatg cagcttatcc atgacgatag cctcactttt     2820

aaagaggaga tcgagaaggc tcaggtttct ggtcaaggtg attctcttca tgaacaaatt     2880

gctgaccttg ctggttcccc tgcaatcaaa aagggtatcc ttcagacagt aaaaatcgtg     2940

gacgagctcg ttaaagtgat ggggcataaa ccggaaaata tcgtaattga aatggcaaga     3000

gaaaaccaaa ctactaccaa aggcctacaa cagtctagag agaggaaaaa aagaatagag     3060

gagggaataa aggaacttga gagtcaaata ctaaaggaga accctgtaga aaatacacaa     3120

ctgcagaacg aaaaattata cttgtactac ttgcaaaatg gccgtgatat gtatgttgac     3180

caagaactgg atattaatag actttctgat tacgatgttg acgcaattgt tccacagagt     3240

tttattaaag atgactccat cgacaacaag gtattaacaa gaagcgttga gaataggggg     3300

aagtcagata acgtaccatc agaagaagtt gtcaaaaaaa tgaagaacta ctggcgtcaa     3360

ttgcttaatg caaaactgat tactcagcga aaatttgata atctcacaaa ggcggagagg     3420

gggggccttt cagaggcaga taaggctggt tttatcaagc gacagttagt tgaaacaagg     3480

caaatcacaa aacatgtcgc cagaatctta gattcgagaa tgaacacaaa gagagataaa     3540

aatgacaagc cgattaggga agtcaaggtt attacattaa aatcgaaatt ggtctctgat     3600

ttccgaaagg actttcaact ttataaggtg agagatatca acaattatca ccatgctcac     3660

gatgcgtacc taaatgccgt cgttggtaca gctcttatca agaaatatcc taagcttgaa     3720

tctgaattcg tttatggaga ctacaaagtg tatgatgttc gaaagatgat agctaaatcc     3780

gaacaagaaa tcggtaaggc tacagcgaaa agattcttct attccaacat tatgaacttt     3840

ttcaaaacag aggtgaagtt agctaatgga gagatcagaa agcgtccttt gatcgagact     3900

aatggtgaga caggtgaagt ggtgtggaat aaggaaaaag attttgctac cgtaaggaag     3960

gtgctagcaa tgccacaagt caacattgtt aaaaagacag aggttcagac gggtggattc     4020

tctaaagagt ctatcctctc taaaagggaa agcgcaaagc taattccgcg taaaaaggga     4080

tgggatacaa gaaagtacgg gggttttgga agcccgaccg ttgcctattc tatccttgtt     4140

gttgctaaag ttgaaaaggg aaaggcaaaa aaactcaaat ctgttaaagt actcgtcggc     4200

attacaataa tggagaaggg ttcgtatgag aaagatccta ttggattcct agaagcaaag     4260

ggttataaag atataaaaaa ggagttgatc ttcaaactcc caaagtacag tcttttcgag     4320

ttagagaatg gtagaagaag gatgttagcc tctgctacgg aactccagaa agcgaatgag     4380

ttagtcctgc cacaacatct tgtaagacta ttatattata ctcaaaatat ttccgctacc     4440

acaggttcga ataacttggg ttacattgag caacatagag aggaattcaa ggaaatattc     4500

gaaaagatta tcgatttttc agaaaaatac atcttgaaga ataaggttaa tagcaatctt     4560

aaatcgtcgt tcgatgagca gttcgctgtc tcagattcaa ttttgttgag caattcgttt     4620

gtttcattgt tgaagtatac gtcatttggt gcctcaggcg gtttcacgtt tctcgatctt     4680

gatgttaaac agggcagatt gagatatcag actgtgaccg aggtattaga tgctactctc     4740

atctatcaaa gtatcactgg gctgtacgag actcgaaccg accttagtca acttggcggg     4800

gattaa                                                                4806


<210>  59
<211>  235
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  35ST from pSAT6

<400>  59
ctgcagagat ccacctagtc tagagtccgc aaaaatcacc agtctctctc tacaaatcta       60

tctctctcta tttttctcca gaataatgtg tgagtagttc ccagataagg gaattagggt      120

tcttataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt      180

tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtgac           235


<210>  60
<211>  5946
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA Plant optimized dScCAS9-FokI expression DNA cassette

<400>  60
caacatgtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa       60

gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc      120

cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac      180

aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt      240

cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg      300

tcttcaaagc aagtggattg atgtgataac atggtggagc acgacacact tgtctactcc      360

aaaaatatca aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg      420

gtaatatccg gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag      480

atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc      540

gttgaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc      600

gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc      660

actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa      720

ggaagttcat ttcatttgga gaggacgtcg agagttctca acacaacata tacaaaacaa      780

acgaatctca agcaatcaag cattctactt ctattgcagc aatttaaatc atttctttta      840

aagcaaaagc aattttctga aaattttcac catttacgaa cgatagtccg gactcagatc      900

tcgagatggc acctaagaag aagcgaaagg ttgggggatc tgggagtcaa ctagttaagt      960

cggagcttga ggagaaaaag tctgagctca ggcataaatt aaagtacgtt ccccatgaat     1020

acattgagct tattgaaatc gctagaaatt ctacacaaga tcgtatcttg gaaatgaagg     1080

tgatggagtt tttcatgaag gtttacgggt acagaggtaa gcaccttgga ggatcgagaa     1140

agccagatgg agctatttac acagtaggta gtcccataga ttatggcgta atcgtagata     1200

ccaaagctta tagtggcggt tataatttgc ctattggtca ggccgatgaa atgcaaaggt     1260

acgtagagga gaatcagaca cgaaataagc atataaaccc taacgagtgg tggaaagttt     1320

atccttcttc agtcacagaa ttcaagttct tgtttgtcag tggacatttc aaaggtaatt     1380

ataaagcgca attaacaaga ctcaatcata taaccaactg taatggtgcg gtcctttctg     1440

tggaagagct cttgattgga ggggagatga ttaaggctgg aactttaacg ctagaggagg     1500

ttcgtcgtaa gtttaataac ggagaaatta atttttccgg atctgaaaca ccgggaacca     1560

gtgagtccgc tactcctgag tctatggaga agaagtactc cattggattg gctattggaa     1620

ctaactccgt gggttgggct gtgattacgg atgattataa ggttccgagc aaaaaattta     1680

aggtactcgg caacaccaac aggaaatcga taaaaaagaa tttgatgggt gctcttttat     1740

tcgatagcgg cgagactgcc gaagctacca ggctcaaacg tacagcaaga cgacgttata     1800

caagaagaaa gaaccgtatt agatacttgc aggagatttt cgcaaacgag atggctaagc     1860

tcgatgactc tttttttcag cgtttggagg agtcctttct tgtcgaagaa gataaaaaaa     1920

acgaaagaca cccgattttt gggaacctgg ctgatgaagt tgcctaccat cgaaattacc     1980

caacaatcta ccatcttaga aagaagcttg cagattctcc ggaaaaagcc gatcttagac     2040

tgatttacct agctttggca cacattatca aattcagagg gcacttcctt atcgagggta     2100

agcttaatgc ggagaattcc gatgttgcta agctttttta tcagctaatc cagacttaca     2160

accaactgtt cgaggaaagc ccacttgacg agattgaggt tgatgctaaa ggcatattgt     2220

cagctcgact tagtaagtcg aagagattgg agaagctcat cgcggttttc cctaatgaga     2280

aaaaaaatgg attgtttggg aacattatag ccttagcttt gggattgacg cctaacttca     2340

aaagcaactt cgatctcact gaagatgcta agcttcaact ttcaaaagac acttatgatg     2400

atgatttaga tgaactcctc ggacaaatag gggatcaata tgccgattta tttagcgctg     2460

cgaagaacct ttcggacgca atcttgctct cagatatcct tagatctaat tctgaagtaa     2520

cgaaagctcc tttatcagcg agcatggtaa aaagatatga tgagcatcat caagatttgg     2580

cacttttgaa gaccttagtg cgacaacaat tcccagagaa atatgctgag atatttaagg     2640

atgacacaaa gaacgggtac gctggatatg tcggcattgg tattaagcat cgtaagcgta     2700

caacgaagtt agctacacaa gaggagtttt ataagtttat taagcctata ttagagaaga     2760

tggacggtgc cgaagagttg ctcgctaaac tgaatcgaga cgatttgcta cgaaagcaga     2820

gaacttttga taatggaagc attcctcacc aaattcacct caaggagctt catgcaattt     2880

taaggagaca ggaggagttt tacccatttc ttaaggaaaa ccgtgaaaaa atagagaaaa     2940

tcttgacgtt ccgtattcca tactatgttg gaccgttagc gagaggaaat tctagattcg     3000

cctggctaac acgaaaatca gaagaggcaa taactccatg gaatttcgag gaagttgtgg     3060

ataagggtgc aagcgcccaa agtttcatcg aacgtatgac taacttcgat gaacagttgc     3120

caaataaaaa ggtattgcct aagcattcgc ttctatacga gtacttcaca gtgtataatg     3180

agctaactaa ggtcaaatat gtaactgagc gaatgcgtaa accagaattc ctgagcggag     3240

aacaaaaaaa ggctattgta gatttgctct ttaagacgaa cagaaaggtt actgttaaac     3300

agcttaagga agattatttt aagaagattg aatgctttga ttctgttgaa atcataggag     3360

tggaggacag gtttaatgct tcattgggca cgtatcatga ccttttgaag attatcaagg     3420

acaaggactt cttagacaac gaggaaaacg aagatatttt ggaggacatt gtcttgacat     3480

tgacactgtt cgaagacagg gaaatgattg aagagaggtt gaaaacatat gcgcatttgt     3540

tcgatgataa ggttatgaag cagctgaagc gtcgacacta taccggctgg ggtagactgt     3600

ctagaaaaat gataaatggt attcgtgata aacagagtgg gaagacaatt ctagattttc     3660

ttaaaagtga tgggttttct aatcgaaatt ttatgcagct tatccatgac gatagcctca     3720

cttttaaaga ggagatcgag aaggctcagg tttctggtca aggtgattct cttcatgaac     3780

aaattgctga ccttgctggt tcccctgcaa tcaaaaaggg tatccttcag acagtaaaaa     3840

tcgtggacga gctcgttaaa gtgatggggc ataaaccgga aaatatcgta attgaaatgg     3900

caagagaaaa ccaaactact accaaaggcc tacaacagtc tagagagagg aaaaaaagaa     3960

tagaggaggg aataaaggaa cttgagagtc aaatactaaa ggagaaccct gtagaaaata     4020

cacaactgca gaacgaaaaa ttatacttgt actacttgca aaatggccgt gatatgtatg     4080

ttgaccaaga actggatatt aatagacttt ctgattacga tgttgacgca attgttccac     4140

agagttttat taaagatgac tccatcgaca acaaggtatt aacaagaagc gttgagaata     4200

gggggaagtc agataacgta ccatcagaag aagttgtcaa aaaaatgaag aactactggc     4260

gtcaattgct taatgcaaaa ctgattactc agcgaaaatt tgataatctc acaaaggcgg     4320

agaggggggg cctttcagag gcagataagg ctggttttat caagcgacag ttagttgaaa     4380

caaggcaaat cacaaaacat gtcgccagaa tcttagattc gagaatgaac acaaagagag     4440

ataaaaatga caagccgatt agggaagtca aggttattac attaaaatcg aaattggtct     4500

ctgatttccg aaaggacttt caactttata aggtgagaga tatcaacaat tatcaccatg     4560

ctcacgatgc gtacctaaat gccgtcgttg gtacagctct tatcaagaaa tatcctaagc     4620

ttgaatctga attcgtttat ggagactaca aagtgtatga tgttcgaaag atgatagcta     4680

aatccgaaca agaaatcggt aaggctacag cgaaaagatt cttctattcc aacattatga     4740

actttttcaa aacagaggtg aagttagcta atggagagat cagaaagcgt cctttgatcg     4800

agactaatgg tgagacaggt gaagtggtgt ggaataagga aaaagatttt gctaccgtaa     4860

ggaaggtgct agcaatgcca caagtcaaca ttgttaaaaa gacagaggtt cagacgggtg     4920

gattctctaa agagtctatc ctctctaaaa gggaaagcgc aaagctaatt ccgcgtaaaa     4980

agggatggga tacaagaaag tacgggggtt ttggaagccc gaccgttgcc tattctatcc     5040

ttgttgttgc taaagttgaa aagggaaagg caaaaaaact caaatctgtt aaagtactcg     5100

tcggcattac aataatggag aagggttcgt atgagaaaga tcctattgga ttcctagaag     5160

caaagggtta taaagatata aaaaaggagt tgatcttcaa actcccaaag tacagtcttt     5220

tcgagttaga gaatggtaga agaaggatgt tagcctctgc tacggaactc cagaaagcga     5280

atgagttagt cctgccacaa catcttgtaa gactattata ttatactcaa aatatttccg     5340

ctaccacagg ttcgaataac ttgggttaca ttgagcaaca tagagaggaa ttcaaggaaa     5400

tattcgaaaa gattatcgat ttttcagaaa aatacatctt gaagaataag gttaatagca     5460

atcttaaatc gtcgttcgat gagcagttcg ctgtctcaga ttcaattttg ttgagcaatt     5520

cgtttgtttc attgttgaag tatacgtcat ttggtgcctc aggcggtttc acgtttctcg     5580

atcttgatgt taaacagggc agattgagat atcagactgt gaccgaggta ttagatgcta     5640

ctctcatcta tcaaagtatc actgggctgt acgagactcg aaccgacctt agtcaacttg     5700

gcggggatta actgcagaga tccacctagt ctagagtccg caaaaatcac cagtctctct     5760

ctacaaatct atctctctct atttttctcc agaataatgt gtgagtagtt cccagataag     5820

ggaattaggg ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat     5880

gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc     5940

agtgac                                                                5946


<210>  61
<211>  711
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Donor sequence comprising PD-mCherry-S encoding ORF

<400>  61
atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcccttcaag       60

gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc      120

cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc      180

ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac      240

cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc      300

gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac      360

ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta      420

atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc      480

gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct      540

gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc      600

aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa      660

cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta a               711


<210>  62
<211>  1711
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DONOR PD-MCHERRY-S for homologous recombination repair pathway

<400>  62
attaaccaaa taaggaaaag aatcatatgg tcatcaattc gtaaatcaaa attttaattt       60

gtgtggtatt taatccatct acatgtttcg taagcaacaa aagagcttgg tctgaaaacc      120

aaacaagacc atatgggcac tcgaatactc cattttgtta tcggctactt ccactagcct      180

cctccttcgc tgcgtctcct gtttctctac ttcacgatta ctcgctaggt aaaagataag      240

atactaaaca actactatag cccctttaat tcaagtccgt ttttttttgt tcttcttctt      300

ttcaattatg tgttaaagat acaaactttt gtctgatttg cttccaccgg tttcacctaa      360

gatactcaat tttcttactt tttgtgtgtt ttgtaattct aattctttta tagcttcaat      420

ttttagattc attgaagcag ttgtgagtta agttggagaa aatggttgtg tttgggaatg      480

tttctgcggc gaatttgcct atggtgagca agggcgagga ggataacatg gccatcatca      540

aggagttcat gcccttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga      600

tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga      660

ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct      720

ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg      780

agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc      840

aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact      900

tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc      960

ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg     1020

acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc     1080

tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca     1140

ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc     1200

tgtacaagta atagctttag ggttcccact tctcaagcgc ttaagacaag aacaaggagg     1260

aggagtactg ctggtccttt gcaggtatct acctttctgg tttttaactt ggagttctcg     1320

aaatttcatc tagtgtataa gtgtaatttt gattaaagag aggaaattgc aggtagtttg     1380

tgtggatatt ccaaggccag agctagagaa cactgtcaat ttcttggaag ctgctagttt     1440

atctgcatcc ttccgtagtg ctcctcgtcc tgctaagcct ttgaaagttg taattgctgg     1500

tgctggtatg tctacaactt catttcttac tgctttgttt gggatctgat ggaattaatg     1560

ttcatgtgat gaagttcttt tggctctcaa ataattgcag gattggctgg attgtcaact     1620

gcaaagtacc tggctgatgc aggccacaaa cctctgttgc ttgaagcaag agatgttctt     1680

ggtggaaagg tactttgatt ggtctcactt t                                    1711


<210>  63
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR Primer2F:

<400>  63
accagaaagg tagatacctg                                                   20


<210>  64
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR Primer2R

<400>  64
atttgcttcc accggtttca c                                                 21


<210>  65
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3241.SP6P SV40NLS F

<400>  65
tacacatacg tatttaggtg acactataga agagaccacc atggccccta agaagaagag       60

a                                                                       61


<210>  66
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3242.CASR

<400>  66
tttttttttt tagtcgcctc cgagctgaga                                        30


<210>  67
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  sgRNA 3243EMX115R

<400>  67
agagccugca cuccuccacc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  68
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  sgRNA 3244EMX115L

<400>  68
ugggacucua cgaguuucua guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  69
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3042

<400>  69
gaaaccatgc ccattc                                                       16


<210>  70
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3240

<400>  70
agccaaggtg gaggagtgca ggctct                                            26


<210>  71
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 1R

<400>  71
tttggtggag gagtgcaggc tct                                               23


<210>  72
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 2R

<400>  72
attggtggag gagtgcaggc tct                                               23


<210>  73
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3R

<400>  73
cttggtggag gagtgcaggc tct                                               23


<210>  74
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 4R

<400>  74
gttggtggag gagtgcaggc tct                                               23


<210>  75
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 5R

<400>  75
tatggtggag gagtgcaggc tct                                               23


<210>  76
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 6R

<400>  76
aatggtggag gagtgcaggc tct                                               23


<210>  77
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 7R

<400>  77
catggtggag gagtgcaggc tct                                               23


<210>  78
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 8R

<400>  78
gatggtggag gagtgcaggc tct                                               23


<210>  79
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 9R

<400>  79
tctggtggag gagtgcaggc tct                                               23


<210>  80
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 10R

<400>  80
actggtggag gagtgcaggc tct                                               23


<210>  81
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 11R

<400>  81
cctggtggag gagtgcaggc tct                                               23


<210>  82
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 12R

<400>  82
gctggtggag gagtgcaggc tct                                               23


<210>  83
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 13R

<400>  83
tgtggtggag gagtgcaggc tct                                               23


<210>  84
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 14R

<400>  84
agtggtggag gagtgcaggc tct                                               23


<210>  85
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 15R

<400>  85
cgtggtggag gagtgcaggc tct                                               23


<210>  86
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 16R

<400>  86
ggtggtggag gagtgcaggc tct                                               23


<210>  87
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 17R

<400>  87
ttaggtggag gagtgcaggc tct                                               23


<210>  88
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 18R

<400>  88
ataggtggag gagtgcaggc tct                                               23


<210>  89
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 19R

<400>  89
ctaggtggag gagtgcaggc tct                                               23


<210>  90
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 20R

<400>  90
gtaggtggag gagtgcaggc tct                                               23


<210>  91
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 21R

<400>  91
taaggtggag gagtgcaggc tct                                               23


<210>  92
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 22R

<400>  92
aaaggtggag gagtgcaggc tct                                               23


<210>  93
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 23R

<400>  93
caaggtggag gagtgcaggc tct                                               23


<210>  94
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 24R

<400>  94
gaaggtggag gagtgcaggc tct                                               23


<210>  95
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 25R

<400>  95
tcaggtggag gagtgcaggc tct                                               23


<210>  96
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 26R

<400>  96
acaggtggag gagtgcaggc tct                                               23


<210>  97
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 27R

<400>  97
ccaggtggag gagtgcaggc tct                                               23


<210>  98
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 28R

<400>  98
gcaggtggag gagtgcaggc tct                                               23


<210>  99
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 29R

<400>  99
tgaggtggag gagtgcaggc tct                                               23


<210>  100
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 30R

<400>  100
agaggtggag gagtgcaggc tct                                               23


<210>  101
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 31R

<400>  101
cgaggtggag gagtgcaggc tct                                               23


<210>  102
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 32R

<400>  102
ggaggtggag gagtgcaggc tct                                               23


<210>  103
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 33R

<400>  103
ttcggtggag gagtgcaggc tct                                               23


<210>  104
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 34R

<400>  104
atcggtggag gagtgcaggc tct                                               23


<210>  105
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 35R

<400>  105
ctcggtggag gagtgcaggc tct                                               23


<210>  106
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 36R

<400>  106
gtcggtggag gagtgcaggc tct                                               23


<210>  107
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 37R

<400>  107
tacggtggag gagtgcaggc tct                                               23


<210>  108
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 38R

<400>  108
aacggtggag gagtgcaggc tct                                               23


<210>  109
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 39R

<400>  109
cacggtggag gagtgcaggc tct                                               23


<210>  110
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 40R

<400>  110
gacggtggag gagtgcaggc tct                                               23


<210>  111
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 41R

<400>  111
tccggtggag gagtgcaggc tct                                               23


<210>  112
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 42R

<400>  112
accggtggag gagtgcaggc tct                                               23


<210>  113
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 43R

<400>  113
cccggtggag gagtgcaggc tct                                               23


<210>  114
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 44R

<400>  114
gccggtggag gagtgcaggc tct                                               23


<210>  115
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 45R

<400>  115
tgcggtggag gagtgcaggc tct                                               23


<210>  116
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 46R

<400>  116
agcggtggag gagtgcaggc tct                                               23


<210>  117
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 47R

<400>  117
cgcggtggag gagtgcaggc tct                                               23


<210>  118
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 48R

<400>  118
ggcggtggag gagtgcaggc tct                                               23


<210>  119
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 49R

<400>  119
ttgggtggag gagtgcaggc tct                                               23


<210>  120
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 50R

<400>  120
atgggtggag gagtgcaggc tct                                               23


<210>  121
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 51R

<400>  121
ctgggtggag gagtgcaggc tct                                               23


<210>  122
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 52R

<400>  122
gtgggtggag gagtgcaggc tct                                               23


<210>  123
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 53R

<400>  123
tagggtggag gagtgcaggc tct                                               23


<210>  124
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 54R

<400>  124
aagggtggag gagtgcaggc tct                                               23


<210>  125
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 55R

<400>  125
cagggtggag gagtgcaggc tct                                               23


<210>  126
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 56R

<400>  126
gagggtggag gagtgcaggc tct                                               23


<210>  127
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 57R

<400>  127
tcgggtggag gagtgcaggc tct                                               23


<210>  128
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 58R

<400>  128
acgggtggag gagtgcaggc tct                                               23


<210>  129
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 59R

<400>  129
ccgggtggag gagtgcaggc tct                                               23


<210>  130
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 60R

<400>  130
gcgggtggag gagtgcaggc tct                                               23


<210>  131
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 61R

<400>  131
tggggtggag gagtgcaggc tct                                               23


<210>  132
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 62R

<400>  132
aggggtggag gagtgcaggc tct                                               23


<210>  133
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 63R

<400>  133
cggggtggag gagtgcaggc tct                                               23


<210>  134
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 64R

<400>  134
gggggtggag gagtgcaggc tct                                               23


<210>  135
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 1L

<400>  135
ttttagaaac tcgtagagtc cca                                               23


<210>  136
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 2L

<400>  136
atttagaaac tcgtagagtc cca                                               23


<210>  137
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 3L

<400>  137
ctttagaaac tcgtagagtc cca                                               23


<210>  138
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 4L

<400>  138
gtttagaaac tcgtagagtc cca                                               23


<210>  139
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 5L

<400>  139
tattagaaac tcgtagagtc cca                                               23


<210>  140
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 6L

<400>  140
aattagaaac tcgtagagtc cca                                               23


<210>  141
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 7L

<400>  141
cattagaaac tcgtagagtc cca                                               23


<210>  142
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 8L

<400>  142
gattagaaac tcgtagagtc cca                                               23


<210>  143
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 9L

<400>  143
tcttagaaac tcgtagagtc cca                                               23


<210>  144
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 10L

<400>  144
acttagaaac tcgtagagtc cca                                               23


<210>  145
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 11L

<400>  145
ccttagaaac tcgtagagtc cca                                               23


<210>  146
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 12L

<400>  146
gcttagaaac tcgtagagtc cca                                               23


<210>  147
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 13L

<400>  147
tgttagaaac tcgtagagtc cca                                               23


<210>  148
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 14L

<400>  148
agttagaaac tcgtagagtc cca                                               23


<210>  149
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 15L

<400>  149
cgttagaaac tcgtagagtc cca                                               23


<210>  150
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 16L

<400>  150
ggttagaaac tcgtagagtc cca                                               23


<210>  151
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 17L

<400>  151
ttatagaaac tcgtagagtc cca                                               23


<210>  152
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 18L

<400>  152
atatagaaac tcgtagagtc cca                                               23


<210>  153
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 19L

<400>  153
ctatagaaac tcgtagagtc cca                                               23


<210>  154
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 20L

<400>  154
gtatagaaac tcgtagagtc cca                                               23


<210>  155
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 21L

<400>  155
taatagaaac tcgtagagtc cca                                               23


<210>  156
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 22L

<400>  156
aaatagaaac tcgtagagtc cca                                               23


<210>  157
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 23L

<400>  157
caatagaaac tcgtagagtc cca                                               23


<210>  158
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 24L

<400>  158
gaatagaaac tcgtagagtc cca                                               23


<210>  159
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 25L

<400>  159
tcatagaaac tcgtagagtc cca                                               23


<210>  160
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 26L

<400>  160
acatagaaac tcgtagagtc cca                                               23


<210>  161
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 27L

<400>  161
ccatagaaac tcgtagagtc cca                                               23


<210>  162
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 28L

<400>  162
gcatagaaac tcgtagagtc cca                                               23


<210>  163
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 29L

<400>  163
tgatagaaac tcgtagagtc cca                                               23


<210>  164
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 30L

<400>  164
agatagaaac tcgtagagtc cca                                               23


<210>  165
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 31L

<400>  165
cgatagaaac tcgtagagtc cca                                               23


<210>  166
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 32L

<400>  166
ggatagaaac tcgtagagtc cca                                               23


<210>  167
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 33L

<400>  167
ttctagaaac tcgtagagtc cca                                               23


<210>  168
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 34L

<400>  168
atctagaaac tcgtagagtc cca                                               23


<210>  169
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 35L

<400>  169
ctctagaaac tcgtagagtc cca                                               23


<210>  170
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 36L

<400>  170
gtctagaaac tcgtagagtc cca                                               23


<210>  171
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 37L

<400>  171
tactagaaac tcgtagagtc cca                                               23


<210>  172
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 38L

<400>  172
aactagaaac tcgtagagtc cca                                               23


<210>  173
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 39L

<400>  173
cactagaaac tcgtagagtc cca                                               23


<210>  174
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 40L

<400>  174
gactagaaac tcgtagagtc cca                                               23


<210>  175
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 41L

<400>  175
tcctagaaac tcgtagagtc cca                                               23


<210>  176
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 42L

<400>  176
acctagaaac tcgtagagtc cca                                               23


<210>  177
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 43L

<400>  177
ccctagaaac tcgtagagtc cca                                               23


<210>  178
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 44L

<400>  178
gcctagaaac tcgtagagtc cca                                               23


<210>  179
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 45L

<400>  179
tgctagaaac tcgtagagtc cca                                               23


<210>  180
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 46L

<400>  180
agctagaaac tcgtagagtc cca                                               23


<210>  181
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 47L

<400>  181
cgctagaaac tcgtagagtc cca                                               23


<210>  182
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 48L

<400>  182
ggctagaaac tcgtagagtc cca                                               23


<210>  183
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 49L

<400>  183
ttgtagaaac tcgtagagtc cca                                               23


<210>  184
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 50L

<400>  184
atgtagaaac tcgtagagtc cca                                               23


<210>  185
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 51L

<400>  185
ctgtagaaac tcgtagagtc cca                                               23


<210>  186
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 52L

<400>  186
gtgtagaaac tcgtagagtc cca                                               23


<210>  187
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 53L

<400>  187
tagtagaaac tcgtagagtc cca                                               23


<210>  188
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 54L

<400>  188
aagtagaaac tcgtagagtc cca                                               23


<210>  189
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 55L

<400>  189
cagtagaaac tcgtagagtc cca                                               23


<210>  190
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 56L

<400>  190
gagtagaaac tcgtagagtc cca                                               23


<210>  191
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 57L

<400>  191
tcgtagaaac tcgtagagtc cca                                               23


<210>  192
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 58L

<400>  192
acgtagaaac tcgtagagtc cca                                               23


<210>  193
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 59L

<400>  193
ccgtagaaac tcgtagagtc cca                                               23


<210>  194
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 60L

<400>  194
gcgtagaaac tcgtagagtc cca                                               23


<210>  195
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 61L

<400>  195
ctggtagaaa ctcgtagagt ccca                                              24


<210>  196
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 62L

<400>  196
aggtagaaac tcgtagagtc cca                                               23


<210>  197
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 63L

<400>  197
cggtagaaac tcgtagagtc cca                                               23


<210>  198
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer 64L

<400>  198
gggtagaaac tcgtagagtc cca                                               23


<210>  199
<211>  60
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Spacer gap N14

<400>  199
ctctagaaac tcgtagagtc ccatgtctgc ggcttccaga gcctgcactc ctccaccttg       60


<210>  200
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Spacer gap N15

<400>  200
ctctagaaac tcgtagagtc ccatgtctgc cggcttccag agcctgcact cctccacctt       60

g                                                                       61


<210>  201
<211>  62
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Spacer gap N16

<400>  201
ctctagaaac tcgtagagtc ccatgtctgc acggcttcca gagcctgcac tcctccacct       60

tg                                                                      62


<210>  202
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Spacer gap N17

<400>  202
ctctagaaac tcgtagagtc ccatgtctgc accggcttcc agagcctgca ctcctccacc       60

ttg                                                                     63


<210>  203
<211>  71
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Spacer gap N25

<400>  203
ctctagaaac tcgtagagtc ccatgtctgc cactgcagtg aggcttccag agcctgcact       60

cctccacctt g                                                            71


<210>  204
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Spacer gap N26

<400>  204
ctctagaaac tcgtagagtc ccatgtctgc cactgtcagt gaggcttcca gagcctgcac       60

tcctccacct tg                                                           72


<210>  205
<211>  73
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Spacer gap N27

<400>  205
ctctagaaac tcgtagagtc ccatgtctgc cactgtacag tgaggcttcc agagcctgca       60

ctcctccacc ttg                                                          73


<210>  206
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI (default linker)

<400>  206

Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser 
1               5                   10                  15      


<210>  207
<211>  25
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker dCasX-FokI (longer linker)

<400>  207

Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Gly 
1               5                   10                  15      


Gly Ser Ala Gly Gly Thr Gly Gly Ser 
            20                  25  


<210>  208
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker dCasX-FokI (shorter linker, small flexible amino acids and
       uncharged)

<400>  208

Gly Gly Ser Ala Gly Gly Ser Gly Ser Gly 
1               5                   10  


<210>  209
<211>  15
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker dCasX-FokI (linker composed of small uncharged amino 
       acids, and contains nuclear localization tag)

<400>  209

Gly Gly Ser Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Thr Gly 
1               5                   10                  15  


<210>  210
<211>  1222
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI (default linker)

<400>  210

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Arg Ile Asn Lys Ile Arg Lys 
225                 230                 235                 240 


Lys Leu Ser Ala Asp Asn Ala Thr Lys Pro Val Ser Arg Ser Gly Pro 
                245                 250                 255     


Met Lys Thr Leu Leu Val Arg Val Met Thr Asp Asp Leu Lys Lys Arg 
            260                 265                 270         


Leu Glu Lys Arg Arg Lys Lys Pro Glu Val Met Pro Gln Val Ile Ser 
        275                 280                 285             


Asn Asn Ala Ala Asn Asn Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys 
    290                 295                 300                 


Met Lys Glu Ala Ile Leu Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp 
305                 310                 315                 320 


His Val Gly Leu Met Cys Lys Phe Ala Gln Pro Ala Ser Lys Lys Ile 
                325                 330                 335     


Asp Gln Asn Lys Leu Lys Pro Glu Met Asp Glu Lys Gly Asn Leu Thr 
            340                 345                 350         


Thr Ala Gly Phe Ala Cys Ser Gln Cys Gly Gln Pro Leu Phe Val Tyr 
        355                 360                 365             


Lys Leu Glu Gln Val Ser Glu Lys Gly Lys Ala Tyr Thr Asn Tyr Phe 
    370                 375                 380                 


Gly Arg Cys Asn Val Ala Glu His Glu Lys Leu Ile Leu Leu Ala Gln 
385                 390                 395                 400 


Leu Lys Pro Glu Lys Asp Ser Asp Glu Ala Val Thr Tyr Ser Leu Gly 
                405                 410                 415     


Ala Ser Lys Phe Gly Gln Arg Ala Leu Asp Phe Tyr Ser Ile His Val 
            420                 425                 430         


Thr Lys Glu Ser Thr His Pro Val Lys Pro Leu Ala Gln Ile Ala Gly 
        435                 440                 445             


Asn Arg Tyr Ala Ser Gly Pro Val Gly Lys Ala Leu Ser Asp Ala Cys 
    450                 455                 460                 


Met Gly Thr Ile Ala Ser Phe Leu Ser Lys Tyr Gln Asp Ile Ile Ile 
465                 470                 475                 480 


Glu His Gln Lys Val Val Lys Gly Asn Gln Lys Arg Leu Glu Ser Leu 
                485                 490                 495     


Arg Glu Leu Ala Gly Lys Glu Asn Leu Glu Tyr Pro Ser Val Thr Leu 
            500                 505                 510         


Pro Pro Gln Pro His Thr Lys Glu Gly Val Asp Ala Tyr Asn Glu Val 
        515                 520                 525             


Ile Ala Arg Val Arg Met Trp Val Asn Leu Asn Leu Trp Gln Lys Leu 
    530                 535                 540                 


Lys Leu Ser Arg Asp Asp Ala Lys Pro Leu Leu Arg Leu Lys Gly Phe 
545                 550                 555                 560 


Pro Ser Phe Pro Val Val Glu Arg Arg Glu Asn Glu Val Asp Trp Trp 
                565                 570                 575     


Asn Thr Ile Asn Glu Val Lys Lys Leu Ile Asp Ala Lys Arg Asp Met 
            580                 585                 590         


Gly Arg Val Phe Trp Ser Gly Val Thr Ala Glu Lys Arg Asn Thr Ile 
        595                 600                 605             


Leu Glu Gly Tyr Asn Tyr Leu Pro Asn Glu Asn Asp His Lys Lys Arg 
    610                 615                 620                 


Glu Gly Ser Leu Glu Asn Pro Lys Lys Pro Ala Lys Arg Gln Phe Gly 
625                 630                 635                 640 


Asp Leu Leu Leu Tyr Leu Glu Lys Lys Tyr Ala Gly Asp Trp Gly Lys 
                645                 650                 655     


Val Phe Asp Glu Ala Trp Glu Arg Ile Asp Lys Lys Ile Ala Gly Leu 
            660                 665                 670         


Thr Ser His Ile Glu Arg Glu Glu Ala Arg Asn Ala Glu Asp Ala Gln 
        675                 680                 685             


Ser Lys Ala Val Leu Thr Asp Trp Leu Arg Ala Lys Ala Ser Phe Val 
    690                 695                 700                 


Leu Glu Arg Leu Lys Glu Met Asp Glu Lys Glu Phe Tyr Ala Cys Glu 
705                 710                 715                 720 


Ile Gln Leu Gln Lys Trp Tyr Gly Asp Leu Arg Gly Asn Pro Phe Ala 
                725                 730                 735     


Val Glu Ala Glu Asn Arg Val Val Asp Ile Ser Ala Phe Ser Ile Gly 
            740                 745                 750         


Ser Asp Gly His Ser Ile Gln Tyr Arg Asn Leu Leu Ala Trp Lys Tyr 
        755                 760                 765             


Leu Glu Asn Gly Lys Arg Glu Phe Tyr Leu Leu Met Asn Tyr Gly Lys 
    770                 775                 780                 


Lys Gly Arg Ile Arg Phe Thr Asp Gly Thr Asp Ile Lys Lys Ser Gly 
785                 790                 795                 800 


Lys Trp Gln Gly Leu Leu Tyr Gly Gly Gly Lys Ala Lys Val Ile Asp 
                805                 810                 815     


Leu Thr Phe Asp Pro Asp Asp Glu Gln Leu Ile Ile Leu Pro Leu Ala 
            820                 825                 830         


Phe Gly Thr Arg Gln Gly Arg Glu Phe Ile Trp Asn Asp Leu Leu Ser 
        835                 840                 845             


Leu Glu Thr Gly Leu Ile Lys Leu Ala Asn Gly Arg Val Ile Glu Lys 
    850                 855                 860                 


Thr Ile Tyr Asn Lys Lys Ile Gly Arg Asp Glu Pro Ala Leu Phe Val 
865                 870                 875                 880 


Ala Leu Thr Phe Glu Arg Arg Glu Val Val Asp Pro Ser Asn Ile Lys 
                885                 890                 895     


Pro Val Asn Leu Ile Gly Val Ala Arg Gly Glu Asn Ile Pro Ala Val 
            900                 905                 910         


Ile Ala Thr Leu Asp Pro Glu Gly Cys Pro Leu Pro Glu Phe Lys Asp 
        915                 920                 925             


Ser Ser Gly Gly Pro Thr Asp Ile Leu Arg Ile Gly Glu Gly Tyr Lys 
    930                 935                 940                 


Glu Lys Gln Arg Ala Ile Gln Ala Ala Lys Glu Val Glu Gln Arg Arg 
945                 950                 955                 960 


Ala Gly Gly Tyr Ser Arg Lys Phe Ala Ser Lys Ser Arg Asn Leu Ala 
                965                 970                 975     


Asp Asp Met Val Arg Asn Ser Ala Arg Asp Leu Phe Tyr His Ala Val 
            980                 985                 990         


Thr His Asp Ala Val Leu Val Phe  Ala Asn Leu Ser Arg  Gly Phe Gly 
        995                 1000                 1005             


Arg Gln  Gly Lys Arg Thr Phe  Met Thr Glu Arg Gln  Tyr Thr Lys 
    1010                 1015                 1020             


Met Glu  Asp Trp Leu Thr Ala  Lys Leu Ala Tyr Glu  Gly Leu Thr 
    1025                 1030                 1035             


Ser Lys  Thr Tyr Leu Ser Lys  Thr Leu Ala Gln Tyr  Thr Ser Lys 
    1040                 1045                 1050             


Thr Cys  Ser Asn Cys Gly Phe  Thr Ile Thr Thr Ala  Asp Tyr Asp 
    1055                 1060                 1065             


Gly Met  Leu Val Arg Leu Lys  Lys Thr Ser Asp Gly  Trp Ala Thr 
    1070                 1075                 1080             


Thr Leu  Asn Asn Lys Glu Leu  Lys Ala Glu Gly Gln  Ile Thr Tyr 
    1085                 1090                 1095             


Tyr Asn  Arg Tyr Lys Arg Gln  Thr Val Glu Lys Glu  Leu Ser Ala 
    1100                 1105                 1110             


Glu Leu  Asp Arg Leu Ser Glu  Glu Ser Gly Asn Asn  Asp Ile Ser 
    1115                 1120                 1125             


Lys Trp  Thr Lys Gly Arg Arg  Asp Glu Ala Leu Phe  Leu Leu Lys 
    1130                 1135                 1140             


Lys Arg  Phe Ser His Gly Ser  Arg Pro Val Gln Glu  Gln Phe Val 
    1145                 1150                 1155             


Cys Leu  Asp Cys Gly His Glu  Val His Ala Ala Glu  Gln Ala Ala 
    1160                 1165                 1170             


Leu Asn  Ile Ala Arg Ser Trp  Leu Phe Leu Asn Ser  Asn Ser Thr 
    1175                 1180                 1185             


Glu Phe  Lys Ser Tyr Lys Ser  Gly Lys Gln Pro Phe  Val Gly Ala 
    1190                 1195                 1200             


Trp Gln  Ala Phe Tyr Lys Arg  Arg Leu Lys Glu Val  Trp Lys Pro 
    1205                 1210                 1215             


Asn Ala  Gly Ala 
    1220         


<210>  211
<211>  1231
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI (longer linker)

<400>  211

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Gly Gly Ser Ala Gly Gly Thr Gly Gly Ser Gly Thr 
225                 230                 235                 240 


Glu Lys Arg Ile Asn Lys Ile Arg Lys Lys Leu Ser Ala Asp Asn Ala 
                245                 250                 255     


Thr Lys Pro Val Ser Arg Ser Gly Pro Met Lys Thr Leu Leu Val Arg 
            260                 265                 270         


Val Met Thr Asp Asp Leu Lys Lys Arg Leu Glu Lys Arg Arg Lys Lys 
        275                 280                 285             


Pro Glu Val Met Pro Gln Val Ile Ser Asn Asn Ala Ala Asn Asn Leu 
    290                 295                 300                 


Arg Met Leu Leu Asp Asp Tyr Thr Lys Met Lys Glu Ala Ile Leu Gln 
305                 310                 315                 320 


Val Tyr Trp Gln Glu Phe Lys Asp Asp His Val Gly Leu Met Cys Lys 
                325                 330                 335     


Phe Ala Gln Pro Ala Ser Lys Lys Ile Asp Gln Asn Lys Leu Lys Pro 
            340                 345                 350         


Glu Met Asp Glu Lys Gly Asn Leu Thr Thr Ala Gly Phe Ala Cys Ser 
        355                 360                 365             


Gln Cys Gly Gln Pro Leu Phe Val Tyr Lys Leu Glu Gln Val Ser Glu 
    370                 375                 380                 


Lys Gly Lys Ala Tyr Thr Asn Tyr Phe Gly Arg Cys Asn Val Ala Glu 
385                 390                 395                 400 


His Glu Lys Leu Ile Leu Leu Ala Gln Leu Lys Pro Glu Lys Asp Ser 
                405                 410                 415     


Asp Glu Ala Val Thr Tyr Ser Leu Gly Ala Ser Lys Phe Gly Gln Arg 
            420                 425                 430         


Ala Leu Asp Phe Tyr Ser Ile His Val Thr Lys Glu Ser Thr His Pro 
        435                 440                 445             


Val Lys Pro Leu Ala Gln Ile Ala Gly Asn Arg Tyr Ala Ser Gly Pro 
    450                 455                 460                 


Val Gly Lys Ala Leu Ser Asp Ala Cys Met Gly Thr Ile Ala Ser Phe 
465                 470                 475                 480 


Leu Ser Lys Tyr Gln Asp Ile Ile Ile Glu His Gln Lys Val Val Lys 
                485                 490                 495     


Gly Asn Gln Lys Arg Leu Glu Ser Leu Arg Glu Leu Ala Gly Lys Glu 
            500                 505                 510         


Asn Leu Glu Tyr Pro Ser Val Thr Leu Pro Pro Gln Pro His Thr Lys 
        515                 520                 525             


Glu Gly Val Asp Ala Tyr Asn Glu Val Ile Ala Arg Val Arg Met Trp 
    530                 535                 540                 


Val Asn Leu Asn Leu Trp Gln Lys Leu Lys Leu Ser Arg Asp Asp Ala 
545                 550                 555                 560 


Lys Pro Leu Leu Arg Leu Lys Gly Phe Pro Ser Phe Pro Val Val Glu 
                565                 570                 575     


Arg Arg Glu Asn Glu Val Asp Trp Trp Asn Thr Ile Asn Glu Val Lys 
            580                 585                 590         


Lys Leu Ile Asp Ala Lys Arg Asp Met Gly Arg Val Phe Trp Ser Gly 
        595                 600                 605             


Val Thr Ala Glu Lys Arg Asn Thr Ile Leu Glu Gly Tyr Asn Tyr Leu 
    610                 615                 620                 


Pro Asn Glu Asn Asp His Lys Lys Arg Glu Gly Ser Leu Glu Asn Pro 
625                 630                 635                 640 


Lys Lys Pro Ala Lys Arg Gln Phe Gly Asp Leu Leu Leu Tyr Leu Glu 
                645                 650                 655     


Lys Lys Tyr Ala Gly Asp Trp Gly Lys Val Phe Asp Glu Ala Trp Glu 
            660                 665                 670         


Arg Ile Asp Lys Lys Ile Ala Gly Leu Thr Ser His Ile Glu Arg Glu 
        675                 680                 685             


Glu Ala Arg Asn Ala Glu Asp Ala Gln Ser Lys Ala Val Leu Thr Asp 
    690                 695                 700                 


Trp Leu Arg Ala Lys Ala Ser Phe Val Leu Glu Arg Leu Lys Glu Met 
705                 710                 715                 720 


Asp Glu Lys Glu Phe Tyr Ala Cys Glu Ile Gln Leu Gln Lys Trp Tyr 
                725                 730                 735     


Gly Asp Leu Arg Gly Asn Pro Phe Ala Val Glu Ala Glu Asn Arg Val 
            740                 745                 750         


Val Asp Ile Ser Ala Phe Ser Ile Gly Ser Asp Gly His Ser Ile Gln 
        755                 760                 765             


Tyr Arg Asn Leu Leu Ala Trp Lys Tyr Leu Glu Asn Gly Lys Arg Glu 
    770                 775                 780                 


Phe Tyr Leu Leu Met Asn Tyr Gly Lys Lys Gly Arg Ile Arg Phe Thr 
785                 790                 795                 800 


Asp Gly Thr Asp Ile Lys Lys Ser Gly Lys Trp Gln Gly Leu Leu Tyr 
                805                 810                 815     


Gly Gly Gly Lys Ala Lys Val Ile Asp Leu Thr Phe Asp Pro Asp Asp 
            820                 825                 830         


Glu Gln Leu Ile Ile Leu Pro Leu Ala Phe Gly Thr Arg Gln Gly Arg 
        835                 840                 845             


Glu Phe Ile Trp Asn Asp Leu Leu Ser Leu Glu Thr Gly Leu Ile Lys 
    850                 855                 860                 


Leu Ala Asn Gly Arg Val Ile Glu Lys Thr Ile Tyr Asn Lys Lys Ile 
865                 870                 875                 880 


Gly Arg Asp Glu Pro Ala Leu Phe Val Ala Leu Thr Phe Glu Arg Arg 
                885                 890                 895     


Glu Val Val Asp Pro Ser Asn Ile Lys Pro Val Asn Leu Ile Gly Val 
            900                 905                 910         


Ala Arg Gly Glu Asn Ile Pro Ala Val Ile Ala Thr Leu Asp Pro Glu 
        915                 920                 925             


Gly Cys Pro Leu Pro Glu Phe Lys Asp Ser Ser Gly Gly Pro Thr Asp 
    930                 935                 940                 


Ile Leu Arg Ile Gly Glu Gly Tyr Lys Glu Lys Gln Arg Ala Ile Gln 
945                 950                 955                 960 


Ala Ala Lys Glu Val Glu Gln Arg Arg Ala Gly Gly Tyr Ser Arg Lys 
                965                 970                 975     


Phe Ala Ser Lys Ser Arg Asn Leu Ala Asp Asp Met Val Arg Asn Ser 
            980                 985                 990         


Ala Arg Asp Leu Phe Tyr His Ala  Val Thr His Asp Ala  Val Leu Val 
        995                 1000                 1005             


Phe Ala  Asn Leu Ser Arg Gly  Phe Gly Arg Gln Gly  Lys Arg Thr 
    1010                 1015                 1020             


Phe Met  Thr Glu Arg Gln Tyr  Thr Lys Met Glu Asp  Trp Leu Thr 
    1025                 1030                 1035             


Ala Lys  Leu Ala Tyr Glu Gly  Leu Thr Ser Lys Thr  Tyr Leu Ser 
    1040                 1045                 1050             


Lys Thr  Leu Ala Gln Tyr Thr  Ser Lys Thr Cys Ser  Asn Cys Gly 
    1055                 1060                 1065             


Phe Thr  Ile Thr Thr Ala Asp  Tyr Asp Gly Met Leu  Val Arg Leu 
    1070                 1075                 1080             


Lys Lys  Thr Ser Asp Gly Trp  Ala Thr Thr Leu Asn  Asn Lys Glu 
    1085                 1090                 1095             


Leu Lys  Ala Glu Gly Gln Ile  Thr Tyr Tyr Asn Arg  Tyr Lys Arg 
    1100                 1105                 1110             


Gln Thr  Val Glu Lys Glu Leu  Ser Ala Glu Leu Asp  Arg Leu Ser 
    1115                 1120                 1125             


Glu Glu  Ser Gly Asn Asn Asp  Ile Ser Lys Trp Thr  Lys Gly Arg 
    1130                 1135                 1140             


Arg Asp  Glu Ala Leu Phe Leu  Leu Lys Lys Arg Phe  Ser His Gly 
    1145                 1150                 1155             


Ser Arg  Pro Val Gln Glu Gln  Phe Val Cys Leu Asp  Cys Gly His 
    1160                 1165                 1170             


Glu Val  His Ala Ala Glu Gln  Ala Ala Leu Asn Ile  Ala Arg Ser 
    1175                 1180                 1185             


Trp Leu  Phe Leu Asn Ser Asn  Ser Thr Glu Phe Lys  Ser Tyr Lys 
    1190                 1195                 1200             


Ser Gly  Lys Gln Pro Phe Val  Gly Ala Trp Gln Ala  Phe Tyr Lys 
    1205                 1210                 1215             


Arg Arg  Leu Lys Glu Val Trp  Lys Pro Asn Ala Gly  Ala 
    1220                 1225                 1230     


<210>  212
<211>  1216
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI (shorter linker, small flexible amino acids and 
       uncharged)

<400>  212

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Gly Gly Ser Ala Gly Gly Ser Gly Ser Gly Gly 
    210                 215                 220                 


Thr Glu Lys Arg Ile Asn Lys Ile Arg Lys Lys Leu Ser Ala Asp Asn 
225                 230                 235                 240 


Ala Thr Lys Pro Val Ser Arg Ser Gly Pro Met Lys Thr Leu Leu Val 
                245                 250                 255     


Arg Val Met Thr Asp Asp Leu Lys Lys Arg Leu Glu Lys Arg Arg Lys 
            260                 265                 270         


Lys Pro Glu Val Met Pro Gln Val Ile Ser Asn Asn Ala Ala Asn Asn 
        275                 280                 285             


Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys Met Lys Glu Ala Ile Leu 
    290                 295                 300                 


Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp His Val Gly Leu Met Cys 
305                 310                 315                 320 


Lys Phe Ala Gln Pro Ala Ser Lys Lys Ile Asp Gln Asn Lys Leu Lys 
                325                 330                 335     


Pro Glu Met Asp Glu Lys Gly Asn Leu Thr Thr Ala Gly Phe Ala Cys 
            340                 345                 350         


Ser Gln Cys Gly Gln Pro Leu Phe Val Tyr Lys Leu Glu Gln Val Ser 
        355                 360                 365             


Glu Lys Gly Lys Ala Tyr Thr Asn Tyr Phe Gly Arg Cys Asn Val Ala 
    370                 375                 380                 


Glu His Glu Lys Leu Ile Leu Leu Ala Gln Leu Lys Pro Glu Lys Asp 
385                 390                 395                 400 


Ser Asp Glu Ala Val Thr Tyr Ser Leu Gly Ala Ser Lys Phe Gly Gln 
                405                 410                 415     


Arg Ala Leu Asp Phe Tyr Ser Ile His Val Thr Lys Glu Ser Thr His 
            420                 425                 430         


Pro Val Lys Pro Leu Ala Gln Ile Ala Gly Asn Arg Tyr Ala Ser Gly 
        435                 440                 445             


Pro Val Gly Lys Ala Leu Ser Asp Ala Cys Met Gly Thr Ile Ala Ser 
    450                 455                 460                 


Phe Leu Ser Lys Tyr Gln Asp Ile Ile Ile Glu His Gln Lys Val Val 
465                 470                 475                 480 


Lys Gly Asn Gln Lys Arg Leu Glu Ser Leu Arg Glu Leu Ala Gly Lys 
                485                 490                 495     


Glu Asn Leu Glu Tyr Pro Ser Val Thr Leu Pro Pro Gln Pro His Thr 
            500                 505                 510         


Lys Glu Gly Val Asp Ala Tyr Asn Glu Val Ile Ala Arg Val Arg Met 
        515                 520                 525             


Trp Val Asn Leu Asn Leu Trp Gln Lys Leu Lys Leu Ser Arg Asp Asp 
    530                 535                 540                 


Ala Lys Pro Leu Leu Arg Leu Lys Gly Phe Pro Ser Phe Pro Val Val 
545                 550                 555                 560 


Glu Arg Arg Glu Asn Glu Val Asp Trp Trp Asn Thr Ile Asn Glu Val 
                565                 570                 575     


Lys Lys Leu Ile Asp Ala Lys Arg Asp Met Gly Arg Val Phe Trp Ser 
            580                 585                 590         


Gly Val Thr Ala Glu Lys Arg Asn Thr Ile Leu Glu Gly Tyr Asn Tyr 
        595                 600                 605             


Leu Pro Asn Glu Asn Asp His Lys Lys Arg Glu Gly Ser Leu Glu Asn 
    610                 615                 620                 


Pro Lys Lys Pro Ala Lys Arg Gln Phe Gly Asp Leu Leu Leu Tyr Leu 
625                 630                 635                 640 


Glu Lys Lys Tyr Ala Gly Asp Trp Gly Lys Val Phe Asp Glu Ala Trp 
                645                 650                 655     


Glu Arg Ile Asp Lys Lys Ile Ala Gly Leu Thr Ser His Ile Glu Arg 
            660                 665                 670         


Glu Glu Ala Arg Asn Ala Glu Asp Ala Gln Ser Lys Ala Val Leu Thr 
        675                 680                 685             


Asp Trp Leu Arg Ala Lys Ala Ser Phe Val Leu Glu Arg Leu Lys Glu 
    690                 695                 700                 


Met Asp Glu Lys Glu Phe Tyr Ala Cys Glu Ile Gln Leu Gln Lys Trp 
705                 710                 715                 720 


Tyr Gly Asp Leu Arg Gly Asn Pro Phe Ala Val Glu Ala Glu Asn Arg 
                725                 730                 735     


Val Val Asp Ile Ser Ala Phe Ser Ile Gly Ser Asp Gly His Ser Ile 
            740                 745                 750         


Gln Tyr Arg Asn Leu Leu Ala Trp Lys Tyr Leu Glu Asn Gly Lys Arg 
        755                 760                 765             


Glu Phe Tyr Leu Leu Met Asn Tyr Gly Lys Lys Gly Arg Ile Arg Phe 
    770                 775                 780                 


Thr Asp Gly Thr Asp Ile Lys Lys Ser Gly Lys Trp Gln Gly Leu Leu 
785                 790                 795                 800 


Tyr Gly Gly Gly Lys Ala Lys Val Ile Asp Leu Thr Phe Asp Pro Asp 
                805                 810                 815     


Asp Glu Gln Leu Ile Ile Leu Pro Leu Ala Phe Gly Thr Arg Gln Gly 
            820                 825                 830         


Arg Glu Phe Ile Trp Asn Asp Leu Leu Ser Leu Glu Thr Gly Leu Ile 
        835                 840                 845             


Lys Leu Ala Asn Gly Arg Val Ile Glu Lys Thr Ile Tyr Asn Lys Lys 
    850                 855                 860                 


Ile Gly Arg Asp Glu Pro Ala Leu Phe Val Ala Leu Thr Phe Glu Arg 
865                 870                 875                 880 


Arg Glu Val Val Asp Pro Ser Asn Ile Lys Pro Val Asn Leu Ile Gly 
                885                 890                 895     


Val Ala Arg Gly Glu Asn Ile Pro Ala Val Ile Ala Thr Leu Asp Pro 
            900                 905                 910         


Glu Gly Cys Pro Leu Pro Glu Phe Lys Asp Ser Ser Gly Gly Pro Thr 
        915                 920                 925             


Asp Ile Leu Arg Ile Gly Glu Gly Tyr Lys Glu Lys Gln Arg Ala Ile 
    930                 935                 940                 


Gln Ala Ala Lys Glu Val Glu Gln Arg Arg Ala Gly Gly Tyr Ser Arg 
945                 950                 955                 960 


Lys Phe Ala Ser Lys Ser Arg Asn Leu Ala Asp Asp Met Val Arg Asn 
                965                 970                 975     


Ser Ala Arg Asp Leu Phe Tyr His Ala Val Thr His Asp Ala Val Leu 
            980                 985                 990         


Val Phe Ala Asn Leu Ser Arg Gly  Phe Gly Arg Gln Gly  Lys Arg Thr 
        995                 1000                 1005             


Phe Met  Thr Glu Arg Gln Tyr  Thr Lys Met Glu Asp  Trp Leu Thr 
    1010                 1015                 1020             


Ala Lys  Leu Ala Tyr Glu Gly  Leu Thr Ser Lys Thr  Tyr Leu Ser 
    1025                 1030                 1035             


Lys Thr  Leu Ala Gln Tyr Thr  Ser Lys Thr Cys Ser  Asn Cys Gly 
    1040                 1045                 1050             


Phe Thr  Ile Thr Thr Ala Asp  Tyr Asp Gly Met Leu  Val Arg Leu 
    1055                 1060                 1065             


Lys Lys  Thr Ser Asp Gly Trp  Ala Thr Thr Leu Asn  Asn Lys Glu 
    1070                 1075                 1080             


Leu Lys  Ala Glu Gly Gln Ile  Thr Tyr Tyr Asn Arg  Tyr Lys Arg 
    1085                 1090                 1095             


Gln Thr  Val Glu Lys Glu Leu  Ser Ala Glu Leu Asp  Arg Leu Ser 
    1100                 1105                 1110             


Glu Glu  Ser Gly Asn Asn Asp  Ile Ser Lys Trp Thr  Lys Gly Arg 
    1115                 1120                 1125             


Arg Asp  Glu Ala Leu Phe Leu  Leu Lys Lys Arg Phe  Ser His Gly 
    1130                 1135                 1140             


Ser Arg  Pro Val Gln Glu Gln  Phe Val Cys Leu Asp  Cys Gly His 
    1145                 1150                 1155             


Glu Val  His Ala Ala Glu Gln  Ala Ala Leu Asn Ile  Ala Arg Ser 
    1160                 1165                 1170             


Trp Leu  Phe Leu Asn Ser Asn  Ser Thr Glu Phe Lys  Ser Tyr Lys 
    1175                 1180                 1185             


Ser Gly  Lys Gln Pro Phe Val  Gly Ala Trp Gln Ala  Phe Tyr Lys 
    1190                 1195                 1200             


Arg Arg  Leu Lys Glu Val Trp  Lys Pro Asn Ala Gly  Ala 
    1205                 1210                 1215     


<210>  213
<211>  1221
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI (linker composed of small uncharged amino acids, and 
       contains nuclear localization tag)

<400>  213

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Gly Gly Ser Ala Pro Lys Lys Lys Arg Lys Val 
    210                 215                 220                 


Gly Gly Thr Gly Gly Thr Glu Lys Arg Ile Asn Lys Ile Arg Lys Lys 
225                 230                 235                 240 


Leu Ser Ala Asp Asn Ala Thr Lys Pro Val Ser Arg Ser Gly Pro Met 
                245                 250                 255     


Lys Thr Leu Leu Val Arg Val Met Thr Asp Asp Leu Lys Lys Arg Leu 
            260                 265                 270         


Glu Lys Arg Arg Lys Lys Pro Glu Val Met Pro Gln Val Ile Ser Asn 
        275                 280                 285             


Asn Ala Ala Asn Asn Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys Met 
    290                 295                 300                 


Lys Glu Ala Ile Leu Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp His 
305                 310                 315                 320 


Val Gly Leu Met Cys Lys Phe Ala Gln Pro Ala Ser Lys Lys Ile Asp 
                325                 330                 335     


Gln Asn Lys Leu Lys Pro Glu Met Asp Glu Lys Gly Asn Leu Thr Thr 
            340                 345                 350         


Ala Gly Phe Ala Cys Ser Gln Cys Gly Gln Pro Leu Phe Val Tyr Lys 
        355                 360                 365             


Leu Glu Gln Val Ser Glu Lys Gly Lys Ala Tyr Thr Asn Tyr Phe Gly 
    370                 375                 380                 


Arg Cys Asn Val Ala Glu His Glu Lys Leu Ile Leu Leu Ala Gln Leu 
385                 390                 395                 400 


Lys Pro Glu Lys Asp Ser Asp Glu Ala Val Thr Tyr Ser Leu Gly Ala 
                405                 410                 415     


Ser Lys Phe Gly Gln Arg Ala Leu Asp Phe Tyr Ser Ile His Val Thr 
            420                 425                 430         


Lys Glu Ser Thr His Pro Val Lys Pro Leu Ala Gln Ile Ala Gly Asn 
        435                 440                 445             


Arg Tyr Ala Ser Gly Pro Val Gly Lys Ala Leu Ser Asp Ala Cys Met 
    450                 455                 460                 


Gly Thr Ile Ala Ser Phe Leu Ser Lys Tyr Gln Asp Ile Ile Ile Glu 
465                 470                 475                 480 


His Gln Lys Val Val Lys Gly Asn Gln Lys Arg Leu Glu Ser Leu Arg 
                485                 490                 495     


Glu Leu Ala Gly Lys Glu Asn Leu Glu Tyr Pro Ser Val Thr Leu Pro 
            500                 505                 510         


Pro Gln Pro His Thr Lys Glu Gly Val Asp Ala Tyr Asn Glu Val Ile 
        515                 520                 525             


Ala Arg Val Arg Met Trp Val Asn Leu Asn Leu Trp Gln Lys Leu Lys 
    530                 535                 540                 


Leu Ser Arg Asp Asp Ala Lys Pro Leu Leu Arg Leu Lys Gly Phe Pro 
545                 550                 555                 560 


Ser Phe Pro Val Val Glu Arg Arg Glu Asn Glu Val Asp Trp Trp Asn 
                565                 570                 575     


Thr Ile Asn Glu Val Lys Lys Leu Ile Asp Ala Lys Arg Asp Met Gly 
            580                 585                 590         


Arg Val Phe Trp Ser Gly Val Thr Ala Glu Lys Arg Asn Thr Ile Leu 
        595                 600                 605             


Glu Gly Tyr Asn Tyr Leu Pro Asn Glu Asn Asp His Lys Lys Arg Glu 
    610                 615                 620                 


Gly Ser Leu Glu Asn Pro Lys Lys Pro Ala Lys Arg Gln Phe Gly Asp 
625                 630                 635                 640 


Leu Leu Leu Tyr Leu Glu Lys Lys Tyr Ala Gly Asp Trp Gly Lys Val 
                645                 650                 655     


Phe Asp Glu Ala Trp Glu Arg Ile Asp Lys Lys Ile Ala Gly Leu Thr 
            660                 665                 670         


Ser His Ile Glu Arg Glu Glu Ala Arg Asn Ala Glu Asp Ala Gln Ser 
        675                 680                 685             


Lys Ala Val Leu Thr Asp Trp Leu Arg Ala Lys Ala Ser Phe Val Leu 
    690                 695                 700                 


Glu Arg Leu Lys Glu Met Asp Glu Lys Glu Phe Tyr Ala Cys Glu Ile 
705                 710                 715                 720 


Gln Leu Gln Lys Trp Tyr Gly Asp Leu Arg Gly Asn Pro Phe Ala Val 
                725                 730                 735     


Glu Ala Glu Asn Arg Val Val Asp Ile Ser Ala Phe Ser Ile Gly Ser 
            740                 745                 750         


Asp Gly His Ser Ile Gln Tyr Arg Asn Leu Leu Ala Trp Lys Tyr Leu 
        755                 760                 765             


Glu Asn Gly Lys Arg Glu Phe Tyr Leu Leu Met Asn Tyr Gly Lys Lys 
    770                 775                 780                 


Gly Arg Ile Arg Phe Thr Asp Gly Thr Asp Ile Lys Lys Ser Gly Lys 
785                 790                 795                 800 


Trp Gln Gly Leu Leu Tyr Gly Gly Gly Lys Ala Lys Val Ile Asp Leu 
                805                 810                 815     


Thr Phe Asp Pro Asp Asp Glu Gln Leu Ile Ile Leu Pro Leu Ala Phe 
            820                 825                 830         


Gly Thr Arg Gln Gly Arg Glu Phe Ile Trp Asn Asp Leu Leu Ser Leu 
        835                 840                 845             


Glu Thr Gly Leu Ile Lys Leu Ala Asn Gly Arg Val Ile Glu Lys Thr 
    850                 855                 860                 


Ile Tyr Asn Lys Lys Ile Gly Arg Asp Glu Pro Ala Leu Phe Val Ala 
865                 870                 875                 880 


Leu Thr Phe Glu Arg Arg Glu Val Val Asp Pro Ser Asn Ile Lys Pro 
                885                 890                 895     


Val Asn Leu Ile Gly Val Ala Arg Gly Glu Asn Ile Pro Ala Val Ile 
            900                 905                 910         


Ala Thr Leu Asp Pro Glu Gly Cys Pro Leu Pro Glu Phe Lys Asp Ser 
        915                 920                 925             


Ser Gly Gly Pro Thr Asp Ile Leu Arg Ile Gly Glu Gly Tyr Lys Glu 
    930                 935                 940                 


Lys Gln Arg Ala Ile Gln Ala Ala Lys Glu Val Glu Gln Arg Arg Ala 
945                 950                 955                 960 


Gly Gly Tyr Ser Arg Lys Phe Ala Ser Lys Ser Arg Asn Leu Ala Asp 
                965                 970                 975     


Asp Met Val Arg Asn Ser Ala Arg Asp Leu Phe Tyr His Ala Val Thr 
            980                 985                 990         


His Asp Ala Val Leu Val Phe Ala  Asn Leu Ser Arg Gly  Phe Gly Arg 
        995                 1000                 1005             


Gln Gly  Lys Arg Thr Phe Met  Thr Glu Arg Gln Tyr  Thr Lys Met 
    1010                 1015                 1020             


Glu Asp  Trp Leu Thr Ala Lys  Leu Ala Tyr Glu Gly  Leu Thr Ser 
    1025                 1030                 1035             


Lys Thr  Tyr Leu Ser Lys Thr  Leu Ala Gln Tyr Thr  Ser Lys Thr 
    1040                 1045                 1050             


Cys Ser  Asn Cys Gly Phe Thr  Ile Thr Thr Ala Asp  Tyr Asp Gly 
    1055                 1060                 1065             


Met Leu  Val Arg Leu Lys Lys  Thr Ser Asp Gly Trp  Ala Thr Thr 
    1070                 1075                 1080             


Leu Asn  Asn Lys Glu Leu Lys  Ala Glu Gly Gln Ile  Thr Tyr Tyr 
    1085                 1090                 1095             


Asn Arg  Tyr Lys Arg Gln Thr  Val Glu Lys Glu Leu  Ser Ala Glu 
    1100                 1105                 1110             


Leu Asp  Arg Leu Ser Glu Glu  Ser Gly Asn Asn Asp  Ile Ser Lys 
    1115                 1120                 1125             


Trp Thr  Lys Gly Arg Arg Asp  Glu Ala Leu Phe Leu  Leu Lys Lys 
    1130                 1135                 1140             


Arg Phe  Ser His Gly Ser Arg  Pro Val Gln Glu Gln  Phe Val Cys 
    1145                 1150                 1155             


Leu Asp  Cys Gly His Glu Val  His Ala Ala Glu Gln  Ala Ala Leu 
    1160                 1165                 1170             


Asn Ile  Ala Arg Ser Trp Leu  Phe Leu Asn Ser Asn  Ser Thr Glu 
    1175                 1180                 1185             


Phe Lys  Ser Tyr Lys Ser Gly  Lys Gln Pro Phe Val  Gly Ala Trp 
    1190                 1195                 1200             


Gln Ala  Phe Tyr Lys Arg Arg  Leu Lys Glu Val Trp  Lys Pro Asn 
    1205                 1210                 1215             


Ala Gly  Ala 
    1220     


<210>  214
<211>  361
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  Human RHO gene exon 1 sequence PRO23HIS mutation

<400>  214
atgaatggca cagaaggccc taacttctac gtgcccttct ccaatgcgac gggtgtggta       60

cgcagccact tcgagtaccc acagtactac ctggctgagc catggcagtt ctccatgctg      120

gccgcctaca tgtttctgct gatcgtgctg ggcttcccca tcaacttcct cacgctctac      180

gtcaccgtcc agcacaagaa gctgcgcacg cctctcaact acatcctgct caacctagcc      240

gtggctgacc tcttcatggt cctaggtggc ttcaccagca ccctctacac ctctctgcat      300

ggatacttcg tcttcgggcc cacaggatgc aatttggagg gcttctttgc caccctgggc      360

g                                                                      361


<210>  215
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI RHO ASL1 SCNA

<400>  215
cauuggagaa gggcacguag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  216
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI RHO SR1 SCNA

<400>  216
gcagccacuu cgaguaccca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  217
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI RHO ASL2 SCNA

<400>  217
uguggguacu cgaaguggcu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  218
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI RHO SR2 SCNA

<400>  218
gccauggcag uucuccaugc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  219
<211>  179
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  Human COMP gene exon 13, deletion in ASP 469

<400>  219
ggatggagac ggacatcagg actctcggga caactgtccc acggtgccta acagtgccca       60

ggaggactca gaccacgatg gccagggtga tgcctgcgac gacgacgaca atgacggagt      120

ccctgacagt cgggacaact gccgcctggt gcctaacccc ggccaggagg acgcggaca       179


<210>  220
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI COMP ASL1 SCNA

<400>  220
aucguggucu gaguccuccu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  221
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI COMP SR1 SCNA

<400>  221
ugcgacgacg acgacaauga guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  222
<211>  76
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  Human PDCD1 gene exon 1

<400>  222
atgcagatcc cacaggcgcc ctggccagtc gtctgggcgg tgctacaact gggctggcgg       60

ccaggatggt tcttag                                                       76


<210>  223
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI PDCD1 ASL1 SCNA

<400>  223
ggccagggcg ccugugggau guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  224
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI PDCD1 SR1 SCNA

<400>  224
gctacaactg ggctggcggc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  225
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI PDCD1 ASL2 SCNA

<400>  225
cgacuggcca gggcgccugu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  226
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI PDCD1 SR2 SCNA

<400>  226
cuggcggcca ggaugguucu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  227
<211>  109
<212>  DNA
<213>  Homo sapiens

<400>  227
atggcttgcc ttggatttca gcggcacaag gctcagctga acctggctac caggacctgg       60

ccctgcactc tcctgttttt tcttctcttc atccctgtct tctgcaaag                  109


<210>  228
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI CTL4 ASL1 SCNA

<400>  228
gtgccgctga aatccaaggc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  229
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI CTL4 SR1 SCNA

<400>  229
ctggctacca ggacctggcc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  230
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI CTL4 ASL2 SCNA

<400>  230
caggtcctgg tagccaggtt guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  231
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Guuuuuucuucucuucauccguuuuagagcuagaaauagcaaguuaaaauaaggcuaguccguua
       ucaacuugaaaaaguggcaccgagucggugc

<400>  231
gttttttctt ctcttcatcc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  232
<211>  87
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  Human Siglec-10 gene exon 1

<400>  232
caggcaggcc ctgcgcctcc tatgcggaga tgctactgcc actgctgctg tcctcgctgc       60

tgggcggtga gtgggccaag ggccgga                                           87


<210>  233
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 ASL1 SCNA

<400>  233
gttttttctt ctcttcatcc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  234
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 SR1 SCNA

<400>  234
gttttttctt ctcttcatcc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  235
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 ASL2 SCNA

<400>  235
gcatctccgc ataggaggcg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  236
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 SR2 SCNA

<400>  236
gcatctccgc ataggaggcg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  237
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 ASL3 SCNA

<400>  237
gtagcatctc cgcataggag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  238
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 SR3 SCNA

<400>  238
gtagcatctc cgcataggag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  239
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 ASL4 SCNA

<400>  239
gtggcagtag catctccgca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  240
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 SR4 SCNA

<400>  240
tgctgggcgg tgagtgggcc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  241
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 ASL5 SCNA

<400>  241
cagcagtggc agtagcatct guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  242
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI Siglec-10 SR5 SCNA

<400>  242
ggcggtgagt gggccaaggg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  243
<211>  104
<212>  DNA
<213>  Bos taurus


<220>
<221>  misc_feature
<223>  Bovine ITGB3 gene exon 1

<400>  243
gacgagatgc gagcgcggcg gctctgggca gcggtgctgg tggtgggggc gctggcgggc       60

gtcggcgtcg gaggtgagtg gggctcccgc gcggtgggga ccca                       104


<210>  244
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 ASL1 SCNA

<400>  244
gctgcccaga gccgccgcgc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  245
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 SR1 SCNA

<400>  245
ggcgcuggcg ggcgucggcg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  246
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 ASL2 SCNA

<400>  246
caccgcugcc cagagccgcc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  247
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 SR2 SCNA

<400>  247
cuggcgggcg ucggcgucgg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  248
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 ASL3 SCNA

<400>  248
agcaccgcug cccagagccg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  249
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 SR3 SCNA

<400>  249
ggcgggcguc ggcgucggag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  250
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 ASL4 SCNA

<400>  250
accaccagca ccgcugccca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  251
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 SR4 SCNA

<400>  251
cgucggcguc ggaggugagu guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  252
<211>  96
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 ASL5 SCNA

<400>  252
cgcccccacc accagcaccg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  253
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI ITGB3 SR5 SCNA

<400>  253
gtcggaggtg agtggggctc guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugc                                 96


<210>  254
<211>  2768
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  Homo sapiens rhodopsin (RHO), mRNA, NM_000539

<400>  254
agagtcatcc agctggagcc ctgagtggct gagctcaggc cttcgcagca ttcttgggtg       60

ggagcagcca cgggtcagcc acaagggcca cagccatgaa tggcacagaa ggccctaact      120

tctacgtgcc cttctccaat gcgacgggtg tggtacgcag ccccttcgag tacccacagt      180

actacctggc tgagccatgg cagttctcca tgctggccgc ctacatgttt ctgctgatcg      240

tgctgggctt ccccatcaac ttcctcacgc tctacgtcac cgtccagcac aagaagctgc      300

gcacgcctct caactacatc ctgctcaacc tagccgtggc tgacctcttc atggtcctag      360

gtggcttcac cagcaccctc tacacctctc tgcatggata cttcgtcttc gggcccacag      420

gatgcaattt ggagggcttc tttgccaccc tgggcggtga aattgccctg tggtccttgg      480

tggtcctggc catcgagcgg tacgtggtgg tgtgtaagcc catgagcaac ttccgcttcg      540

gggagaacca tgccatcatg ggcgttgcct tcacctgggt catggcgctg gcctgcgccg      600

cacccccact cgccggctgg tccaggtaca tccccgaggg cctgcagtgc tcgtgtggaa      660

tcgactacta cacgctcaag ccggaggtca acaacgagtc ttttgtcatc tacatgttcg      720

tggtccactt caccatcccc atgattatca tctttttctg ctatgggcag ctcgtcttca      780

ccgtcaagga ggccgctgcc cagcagcagg agtcagccac cacacagaag gcagagaagg      840

aggtcacccg catggtcatc atcatggtca tcgctttcct gatctgctgg gtgccctacg      900

ccagcgtggc attctacatc ttcacccacc agggctccaa cttcggtccc atcttcatga      960

ccatcccagc gttctttgcc aagagcgccg ccatctacaa ccctgtcatc tatatcatga     1020

tgaacaagca gttccggaac tgcatgctca ccaccatctg ctgcggcaag aacccactgg     1080

gtgacgatga ggcctctgct accgtgtcca agacggagac gagccaggtg gccccggcct     1140

aagacctgcc taggactctg tggccgacta taggcgtctc ccatccccta caccttcccc     1200

cagccacagc catcccacca ggagcagcgc ctgtgcagaa tgaacgaagt cacataggct     1260

ccttaatttt tttttttttt ttaagaaata attaatgagg ctcctcactc acctgggaca     1320

gcctgagaag ggacatccac caagacctac tgatctggag tcccacgttc cccaaggcca     1380

gcgggatgtg tgcccctcct cctcccaact catctttcag gaacacgagg attcttgctt     1440

tctggaaaag tgtcccagct tagggataag tgtctagcac agaatggggc acacagtagg     1500

tgcttaataa atgctggatg gatgcaggaa ggaatggagg aatgaatggg aagggagaac     1560

atatctatcc tctcagaccc tcgcagcagc agcaactcat acttggctaa tgatatggag     1620

cagttgtttt tccctccctg ggcctcactt tcttctccta taaaatggaa atcccagatc     1680

cctggtcctg ccgacacgca gctactgaga agaccaaaag aggtgtgtgt gtgtctatgt     1740

gtgtgtttca gcactttgta aatagcaaga agctgtacag attctagtta atgttgtgaa     1800

taacatcaat taatgtaact agttaattac tatgattatc acctcctgat agtgaacatt     1860

ttgagattgg gcattcagat gatggggttt cacccaacct tggggcaggt ttttaaaaat     1920

tagctaggca tcaaggccag accagggctg ggggttgggc tgtaggcagg gacagtcaca     1980

ggaatgcaga atgcagtcat cagacctgaa aaaacaacac tgggggaggg ggacggtgaa     2040

ggccaagttc ccaatgaggg tgagattggg cctggggtct cacccctagt gtggggcccc     2100

aggtcccgtg cctccccttc ccaatgtggc ctatggagag acaggccttt ctctcagcct     2160

ctggaagcca cctgctcttt tgctctagca cctgggtccc agcatctaga gcatggagcc     2220

tctagaagcc atgctcaccc gcccacattt aattaacagc tgagtccctg atgtcatcct     2280

tatctcgaag agcttagaaa caaagagtgg gaaattccac tgggcctacc ttccttgggg     2340

atgttcatgg gccccagttt ccagtttccc ttgccagaca agcccatctt cagcagttgc     2400

tagtccattc tccattctgg agaatctgct ccaaaaagct ggccacatct ctgaggtgtc     2460

agaattaagc tgcctcagta actgctcccc cttctccata taagcaaagc cagaagctct     2520

agctttaccc agctctgcct ggagactaag gcaaattggg ccattaaaag ctcagctcct     2580

atgttggtat taacggtggt gggttttgtt gctttcacac tctatccaca ggatagattg     2640

aaactgccag cttccacctg atccctgacc ctgggatggc tggattgagc aatgagcaga     2700

gccaagcagc acagagtccc ctggggctag aggtggagga ggcagtcctg ggaatgggaa     2760

aaacccca                                                              2768


<210>  255
<211>  348
<212>  PRT
<213>  Homo sapiens


<220>
<221>  MISC_FEATURE
<223>  Homo sapiens rhodopsin (RHO), amino acid, NM_000539

<400>  255

Met Asn Gly Thr Glu Gly Pro Asn Phe Tyr Val Pro Phe Ser Asn Ala 
1               5                   10                  15      


Thr Gly Val Val Arg Ser Pro Phe Glu Tyr Pro Gln Tyr Tyr Leu Ala 
            20                  25                  30          


Glu Pro Trp Gln Phe Ser Met Leu Ala Ala Tyr Met Phe Leu Leu Ile 
        35                  40                  45              


Val Leu Gly Phe Pro Ile Asn Phe Leu Thr Leu Tyr Val Thr Val Gln 
    50                  55                  60                  


His Lys Lys Leu Arg Thr Pro Leu Asn Tyr Ile Leu Leu Asn Leu Ala 
65                  70                  75                  80  


Val Ala Asp Leu Phe Met Val Leu Gly Gly Phe Thr Ser Thr Leu Tyr 
                85                  90                  95      


Thr Ser Leu His Gly Tyr Phe Val Phe Gly Pro Thr Gly Cys Asn Leu 
            100                 105                 110         


Glu Gly Phe Phe Ala Thr Leu Gly Gly Glu Ile Ala Leu Trp Ser Leu 
        115                 120                 125             


Val Val Leu Ala Ile Glu Arg Tyr Val Val Val Cys Lys Pro Met Ser 
    130                 135                 140                 


Asn Phe Arg Phe Gly Glu Asn His Ala Ile Met Gly Val Ala Phe Thr 
145                 150                 155                 160 


Trp Val Met Ala Leu Ala Cys Ala Ala Pro Pro Leu Ala Gly Trp Ser 
                165                 170                 175     


Arg Tyr Ile Pro Glu Gly Leu Gln Cys Ser Cys Gly Ile Asp Tyr Tyr 
            180                 185                 190         


Thr Leu Lys Pro Glu Val Asn Asn Glu Ser Phe Val Ile Tyr Met Phe 
        195                 200                 205             


Val Val His Phe Thr Ile Pro Met Ile Ile Ile Phe Phe Cys Tyr Gly 
    210                 215                 220                 


Gln Leu Val Phe Thr Val Lys Glu Ala Ala Ala Gln Gln Gln Glu Ser 
225                 230                 235                 240 


Ala Thr Thr Gln Lys Ala Glu Lys Glu Val Thr Arg Met Val Ile Ile 
                245                 250                 255     


Met Val Ile Ala Phe Leu Ile Cys Trp Val Pro Tyr Ala Ser Val Ala 
            260                 265                 270         


Phe Tyr Ile Phe Thr His Gln Gly Ser Asn Phe Gly Pro Ile Phe Met 
        275                 280                 285             


Thr Ile Pro Ala Phe Phe Ala Lys Ser Ala Ala Ile Tyr Asn Pro Val 
    290                 295                 300                 


Ile Tyr Ile Met Met Asn Lys Gln Phe Arg Asn Cys Met Leu Thr Thr 
305                 310                 315                 320 


Ile Cys Cys Gly Lys Asn Pro Leu Gly Asp Asp Glu Ala Ser Ala Thr 
                325                 330                 335     


Val Ser Lys Thr Glu Thr Ser Gln Val Ala Pro Ala 
            340                 345             


<210>  256
<211>  194
<212>  PRT
<213>  Flavobacterium okeanokoites


<220>
<221>  MISC_FEATURE
<223>  FokI

<400>  256

Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
1               5                   10                  15      


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
            20                  25                  30          


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
        35                  40                  45              


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
    50                  55                  60                  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
65                  70                  75                  80  


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
                85                  90                  95      


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
            100                 105                 110         


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
        115                 120                 125             


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
    130                 135                 140                 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
145                 150                 155                 160 


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
                165                 170                 175     


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
            180                 185                 190         


Asn Phe 
        


<210>  257
<211>  1368
<212>  PRT
<213>  Streptococcus pyogenes


<220>
<221>  MISC_FEATURE
<223>  Sp Cas9

<400>  257

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


<210>  258
<211>  1375
<212>  PRT
<213>  Streptococcus canis


<220>
<221>  MISC_FEATURE
<223>  Sc Cas9

<400>  258

Met Glu Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu Met 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Arg 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp Ser 
                85                  90                  95      


Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala Tyr 
        115                 120                 125             


His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp 
    130                 135                 140                 


Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn Ala 
                165                 170                 175     


Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp Ala 
        195                 200                 205             


Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu Lys 
    210                 215                 220                 


Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys 
                325                 330                 335     


Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys 
            340                 345                 350         


Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys 
        355                 360                 365             


His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys 
    370                 375                 380                 


Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu 
385                 390                 395                 400 


Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp 
                405                 410                 415     


Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile 
            420                 425                 430         


Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu 
        435                 440                 445             


Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro 
    450                 455                 460                 


Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu 
465                 470                 475                 480 


Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala 
                485                 490                 495     


Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu 
            500                 505                 510         


Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe 
        515                 520                 525             


Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met 
    530                 535                 540                 


Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp 
545                 550                 555                 560 


Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu 
                565                 570                 575     


Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly 
            580                 585                 590         


Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu 
        595                 600                 605             


Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp 
    610                 615                 620                 


Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu 
625                 630                 635                 640 


Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys 
                645                 650                 655     


Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu 
            660                 665                 670         


Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr 
        675                 680                 685             


Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met 
    690                 695                 700                 


Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys 
705                 710                 715                 720 


Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp 
                725                 730                 735     


Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys 
            740                 745                 750         


Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn Ile 
        755                 760                 765             


Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu Gln 
    770                 775                 780                 


Gln Ser Arg Glu Arg Lys Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu 
785                 790                 795                 800 


Glu Ser Gln Ile Leu Lys Glu Asn Pro Val Glu Asn Thr Gln Leu Gln 
                805                 810                 815     


Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr 
            820                 825                 830         


Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp 
        835                 840                 845             


His Ile Val Pro Gln Ser Phe Ile Lys Asp Asp Ser Ile Asp Asn Lys 
    850                 855                 860                 


Val Leu Thr Arg Ser Val Glu Asn Arg Gly Lys Ser Asp Asn Val Pro 
865                 870                 875                 880 


Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu 
                885                 890                 895     


Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala 
            900                 905                 910         


Glu Arg Gly Gly Leu Ser Glu Ala Asp Lys Ala Gly Phe Ile Lys Arg 
        915                 920                 925             


Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Arg Ile Leu 
    930                 935                 940                 


Asp Ser Arg Met Asn Thr Lys Arg Asp Lys Asn Asp Lys Pro Ile Arg 
945                 950                 955                 960 


Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg 
                965                 970                 975     


Lys Asp Phe Gln Leu Tyr Lys Val Arg Asp Ile Asn Asn Tyr His His 
            980                 985                 990         


Ala His Asp Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
        995                 1000                 1005             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1010                 1015                 1020             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1025                 1030                 1035             


Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1040                 1045                 1050             


Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1055                 1060                 1065             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp 
    1070                 1075                 1080             


Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg Lys Val  Leu Ala Met 
    1085                 1090                 1095             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1100                 1105                 1110             


Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu 
    1115                 1120                 1125             


Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe 
    1130                 1135                 1140             


Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val 
    1145                 1150                 1155             


Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser Val Lys  Val Leu Val 
    1160                 1165                 1170             


Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile 
    1175                 1180                 1185             


Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu 
    1190                 1195                 1200             


Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1205                 1210                 1215             


Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn 
    1220                 1225                 1230             


Glu Leu  Val Leu Pro Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr 
    1235                 1240                 1245             


Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile 
    1250                 1255                 1260             


Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile 
    1265                 1270                 1275             


Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn 
    1280                 1285                 1290             


Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile 
    1295                 1300                 1305             


Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe 
    1310                 1315                 1320             


Gly Ala  Ser Gly Gly Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln 
    1325                 1330                 1335             


Gly Arg  Leu Arg Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1340                 1345                 1350             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1355                 1360                 1365             


Leu Ser  Gln Leu Gly Gly Asp  
    1370                 1375 


<210>  259
<211>  46
<212>  PRT
<213>  Escherichia coli


<220>
<221>  MISC_FEATURE
<223>  E. coli Lac repressor fragment 1-46

<400>  259

Met Lys Pro Val Thr Leu Tyr Asp Val Ala Glu Tyr Ala Gly Val Ser 
1               5                   10                  15      


Tyr Gln Thr Val Ser Arg Val Val Asn Gln Ala Ser His Val Ser Ala 
            20                  25                  30          


Lys Thr Arg Glu Lys Val Glu Ala Ala Met Ala Glu Leu Asn 
        35                  40                  45      


<210>  260
<211>  52
<212>  PRT
<213>  Human immunodeficiency virus


<220>
<221>  MISC_FEATURE
<223>  HIV integrase fragment 219-270

<400>  260

Met Ile Gln Asn Phe Arg Val Tyr Tyr Arg Asp Ser Arg Asn Pro Leu 
1               5                   10                  15      


Trp Lys Gly Pro Ala Lys Leu Leu Trp Lys Gly Glu Gly Ala Val Val 
            20                  25                  30          


Ile Gln Asp Asn Ser Asp Ile Lys Val Val Pro Arg Arg Lys Ala Lys 
        35                  40                  45              


Ile Ile Arg Asp 
    50          


<210>  261
<211>  64
<212>  PRT
<213>  Sulfolobus solfataricus


<220>
<221>  MISC_FEATURE
<223>  Residues 1 to 64 of the Sso7D DNA-binding protein of Sulfolobus 
       solfataricus

<400>  261

Met Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp 
1               5                   10                  15      


Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly Lys Met Ile Ser Phe 
            20                  25                  30          


Thr Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg Gly Ala Val Ser Glu 
        35                  40                  45              


Lys Asp Ala Pro Lys Glu Leu Leu Gln Met Leu Glu Lys Gln Lys Lys 
    50                  55                  60                  


<210>  262
<211>  64
<212>  PRT
<213>  Sulfolobus tokodaii


<220>
<221>  MISC_FEATURE
<223>  residues 1 to 64 of the Sto7D DNA-binding protein from Sulfolobus
       tokodaii

<400>  262

Met Val Thr Val Lys Phe Lys Tyr Lys Gly Glu Glu Lys Glu Val Asp 
1               5                   10                  15      


Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly Lys Met Ile Ser Phe 
            20                  25                  30          


Thr Tyr Asp Asp Asn Gly Lys Thr Gly Arg Gly Ala Val Ser Glu Lys 
        35                  40                  45              


Asp Ala Pro Lys Glu Leu Leu Gln Met Leu Glu Lys Ser Gly Lys Lys 
    50                  55                  60                  


<210>  263
<211>  74
<212>  PRT
<213>  Arabidopsis thaliana


<220>
<221>  MISC_FEATURE
<223>  StkC domain comprise residues 232-305 of Arabidopsis MBD7 
       methyl-CpG-binding domain

<400>  263

Ser Val Ser Met Val His Ser Glu Arg Leu Pro Leu Leu Met Asn Arg 
1               5                   10                  15      


Asn Gly Ile Arg Phe Gln Ser Glu Val Ile Asp Pro Asn Pro Pro Lys 
            20                  25                  30          


Lys Val Lys Trp Val Leu Thr Gly Ser Gly Gly Asn Met Phe Thr Ala 
        35                  40                  45              


Asn Val Arg Gly Ser Asn Val Ser Ser Leu Val Lys His Thr Trp Ser 
    50                  55                  60                  


Glu Ala Phe Val Ser Leu Ile Glu Asp Arg 
65                  70                  


<210>  264
<211>  73
<212>  PRT
<213>  Homo sapiens


<220>
<221>  MISC_FEATURE
<223>  human HMG4. Comprising residues 2 to 79 of human HMGB4

<400>  264

Ser Asp Lys Pro Lys Arg Pro Leu Ser Ala Tyr Met Leu Trp Leu Asn 
1               5                   10                  15      


Ser Ala Arg Glu Ser Ile Lys Arg Glu Asn Pro Gly Ile Lys Val Thr 
            20                  25                  30          


Glu Val Ala Lys Arg Gly Gly Glu Leu Trp Arg Ala Met Lys Asp Lys 
        35                  40                  45              


Ser Glu Trp Glu Ala Lys Ala Ala Lys Ala Lys Asp Asp Tyr Asp Arg 
    50                  55                  60                  


Ala Val Lys Glu Phe Glu Ala Asn Gly 
65                  70              


<210>  265
<211>  25
<212>  PRT
<213>  Mus musculus


<220>
<221>  MISC_FEATURE
<223>  Testis zinc finger 3 (non-specific zinc finger)

<400>  265

Ala Pro Tyr Arg Cys Pro Cys Arg Ala Gly Cys Pro Ser Leu Ala Ser 
1               5                   10                  15      


Met Gln Ala His Met Arg Gly His Ser 
            20                  25  


<210>  266
<211>  1339
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-StkC2

<400>  266

Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu Met 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Arg 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp Ser 
                85                  90                  95      


Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala Tyr 
        115                 120                 125             


His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp 
    130                 135                 140                 


Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn Ala 
                165                 170                 175     


Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp Ala 
        195                 200                 205             


Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu Lys 
    210                 215                 220                 


Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys 
                325                 330                 335     


Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys 
            340                 345                 350         


Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys 
        355                 360                 365             


His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys 
    370                 375                 380                 


Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu 
385                 390                 395                 400 


Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp 
                405                 410                 415     


Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile 
            420                 425                 430         


Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu 
        435                 440                 445             


Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro 
    450                 455                 460                 


Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu 
465                 470                 475                 480 


Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala 
                485                 490                 495     


Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu 
            500                 505                 510         


Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe 
        515                 520                 525             


Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met 
    530                 535                 540                 


Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp 
545                 550                 555                 560 


Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu 
                565                 570                 575     


Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly 
            580                 585                 590         


Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu 
        595                 600                 605             


Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp 
    610                 615                 620                 


Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu 
625                 630                 635                 640 


Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys 
                645                 650                 655     


Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu 
            660                 665                 670         


Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr 
        675                 680                 685             


Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met 
    690                 695                 700                 


Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys 
705                 710                 715                 720 


Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp 
                725                 730                 735     


Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys 
            740                 745                 750         


Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn Ile 
        755                 760                 765             


Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu Gln 
    770                 775                 780                 


Gln Ser Arg Glu Arg Lys Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu 
785                 790                 795                 800 


Glu Ser Gln Ile Leu Lys Glu Asn Pro Val Glu Asn Thr Gln Leu Gln 
                805                 810                 815     


Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr 
            820                 825                 830         


Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp 
        835                 840                 845             


Ala Ile Val Pro Gln Ser Phe Ile Lys Asp Asp Ser Ile Asp Asn Lys 
    850                 855                 860                 


Val Leu Thr Arg Ser Val Glu Asn Arg Gly Lys Ser Asp Asn Val Pro 
865                 870                 875                 880 


Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu 
                885                 890                 895     


Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala 
            900                 905                 910         


Glu Arg Gly Gly Leu Ser Glu Ala Asp Lys Ala Gly Phe Ile Lys Arg 
        915                 920                 925             


Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Arg Ile Leu 
    930                 935                 940                 


Asp Ser Arg Met Asn Thr Lys Arg Asp Lys Asn Asp Lys Pro Ile Arg 
945                 950                 955                 960 


Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg 
                965                 970                 975     


Lys Asp Phe Gln Leu Tyr Lys Val Arg Asp Ile Asn Asn Tyr His His 
            980                 985                 990         


Ala His Asp Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
        995                 1000                 1005             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1010                 1015                 1020             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1025                 1030                 1035             


Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1040                 1045                 1050             


Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1055                 1060                 1065             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp 
    1070                 1075                 1080             


Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg Lys Val  Leu Ala Met 
    1085                 1090                 1095             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1100                 1105                 1110             


Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu 
    1115                 1120                 1125             


Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe 
    1130                 1135                 1140             


Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val 
    1145                 1150                 1155             


Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser Val Lys  Val Leu Val 
    1160                 1165                 1170             


Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile 
    1175                 1180                 1185             


Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu 
    1190                 1195                 1200             


Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1205                 1210                 1215             


Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Ser 
    1220                 1225                 1230             


Val Ser  Met Val His Ser Glu  Arg Leu Pro Leu Leu  Met Asn Arg 
    1235                 1240                 1245             


Asn Gly  Ile Arg Phe Gln Ser  Glu Val Ile Asp Pro  Asn Pro Pro 
    1250                 1255                 1260             


Lys Lys  Val Lys Trp Val Leu  Thr Gly Ser Gly Gly  Asn Met Phe 
    1265                 1270                 1275             


Thr Ala  Asn Val Arg Gly Ser  Asn Val Ser Ser Leu  Val Lys His 
    1280                 1285                 1290             


Thr Trp  Ser Glu Ala Phe Val  Ser Leu Ile Glu Asp  Arg Tyr Gln 
    1295                 1300                 1305             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1310                 1315                 1320             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1325                 1330                 1335             


Asp 
    


<210>  267
<211>  1435
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-HMGB4

<400>  267

Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu Met 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Arg 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp Ser 
                85                  90                  95      


Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala Tyr 
        115                 120                 125             


His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp 
    130                 135                 140                 


Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn Ala 
                165                 170                 175     


Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp Ala 
        195                 200                 205             


Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu Lys 
    210                 215                 220                 


Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys 
                325                 330                 335     


Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys 
            340                 345                 350         


Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys 
        355                 360                 365             


His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys 
    370                 375                 380                 


Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu 
385                 390                 395                 400 


Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp 
                405                 410                 415     


Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile 
            420                 425                 430         


Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu 
        435                 440                 445             


Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro 
    450                 455                 460                 


Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu 
465                 470                 475                 480 


Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala 
                485                 490                 495     


Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu 
            500                 505                 510         


Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe 
        515                 520                 525             


Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met 
    530                 535                 540                 


Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp 
545                 550                 555                 560 


Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu 
                565                 570                 575     


Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly 
            580                 585                 590         


Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu 
        595                 600                 605             


Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp 
    610                 615                 620                 


Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu 
625                 630                 635                 640 


Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys 
                645                 650                 655     


Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu 
            660                 665                 670         


Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr 
        675                 680                 685             


Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met 
    690                 695                 700                 


Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys 
705                 710                 715                 720 


Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp 
                725                 730                 735     


Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys 
            740                 745                 750         


Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn Ile 
        755                 760                 765             


Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu Gln 
    770                 775                 780                 


Gln Ser Arg Glu Arg Lys Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu 
785                 790                 795                 800 


Glu Ser Gln Ile Leu Lys Glu Asn Pro Val Glu Asn Thr Gln Leu Gln 
                805                 810                 815     


Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr 
            820                 825                 830         


Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp 
        835                 840                 845             


Ala Ile Val Pro Gln Ser Phe Ile Lys Asp Asp Ser Ile Asp Asn Lys 
    850                 855                 860                 


Val Leu Thr Arg Ser Val Glu Asn Arg Gly Lys Ser Asp Asn Val Pro 
865                 870                 875                 880 


Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu 
                885                 890                 895     


Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala 
            900                 905                 910         


Glu Arg Gly Gly Leu Ser Glu Ala Asp Lys Ala Gly Phe Ile Lys Arg 
        915                 920                 925             


Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Arg Ile Leu 
    930                 935                 940                 


Asp Ser Arg Met Asn Thr Lys Arg Asp Lys Asn Asp Lys Pro Ile Arg 
945                 950                 955                 960 


Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg 
                965                 970                 975     


Lys Asp Phe Gln Leu Tyr Lys Val Arg Asp Ile Asn Asn Tyr His His 
            980                 985                 990         


Ala His Asp Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
        995                 1000                 1005             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1010                 1015                 1020             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1025                 1030                 1035             


Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1040                 1045                 1050             


Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1055                 1060                 1065             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp 
    1070                 1075                 1080             


Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg Lys Val  Leu Ala Met 
    1085                 1090                 1095             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1100                 1105                 1110             


Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu 
    1115                 1120                 1125             


Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe 
    1130                 1135                 1140             


Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val 
    1145                 1150                 1155             


Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser Val Lys  Val Leu Val 
    1160                 1165                 1170             


Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile 
    1175                 1180                 1185             


Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu 
    1190                 1195                 1200             


Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1205                 1210                 1215             


Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn 
    1220                 1225                 1230             


Glu Leu  Val Leu Pro Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr 
    1235                 1240                 1245             


Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile 
    1250                 1255                 1260             


Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile 
    1265                 1270                 1275             


Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn 
    1280                 1285                 1290             


Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile 
    1295                 1300                 1305             


Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe 
    1310                 1315                 1320             


Gly Ala  Ser Gly Gly Phe Ser  Asp Lys Pro Lys Arg  Pro Leu Ser 
    1325                 1330                 1335             


Ala Tyr  Met Leu Trp Leu Asn  Ser Ala Arg Glu Ser  Ile Lys Arg 
    1340                 1345                 1350             


Glu Asn  Pro Gly Ile Lys Val  Thr Glu Val Ala Lys  Arg Gly Gly 
    1355                 1360                 1365             


Glu Leu  Trp Arg Ala Met Lys  Asp Lys Ser Glu Trp  Glu Ala Lys 
    1370                 1375                 1380             


Ala Ala  Lys Ala Lys Asp Asp  Tyr Asp Arg Ala Val  Lys Glu Phe 
    1385                 1390                 1395             


Glu Ala  Asn Gly Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1400                 1405                 1410             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1415                 1420                 1425             


Leu Ser  Gln Leu Gly Gly Asp  
    1430                 1435 


<210>  268
<211>  1375
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Ancestral Cas9

<400>  268

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Met 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp 
    130                 135                 140                 


Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Ala 
                165                 170                 175     


Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys 
                325                 330                 335     


Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Ser 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Ile Gly Ile Lys 
        355                 360                 365             


His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu Glu Phe Tyr Lys 
    370                 375                 380                 


Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu 
385                 390                 395                 400 


Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp 
                405                 410                 415     


Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile 
            420                 425                 430         


Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu 
        435                 440                 445             


Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro 
    450                 455                 460                 


Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Asp 
465                 470                 475                 480 


Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala 
                485                 490                 495     


Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu 
            500                 505                 510         


Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe 
        515                 520                 525             


Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met 
    530                 535                 540                 


Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu Ala Ile Val Asp 
545                 550                 555                 560 


Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu 
                565                 570                 575     


Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly 
            580                 585                 590         


Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu 
        595                 600                 605             


Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp 
    610                 615                 620                 


Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu 
625                 630                 635                 640 


Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys 
                645                 650                 655     


Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu 
            660                 665                 670         


Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr 
        675                 680                 685             


Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met 
    690                 695                 700                 


Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys 
705                 710                 715                 720 


Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp 
                725                 730                 735     


Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys 
            740                 745                 750         


Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn Ile 
        755                 760                 765             


Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys 
    770                 775                 780                 


Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu 
785                 790                 795                 800 


Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln 
                805                 810                 815     


Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Lys Asp Met Tyr 
            820                 825                 830         


Thr Gly Asp Glu Leu Asp Ile Asn Arg Leu Tyr Gln Tyr Asp Val Asp 
        835                 840                 845             


His Ile Ile Pro Gln Ser Phe Ile Lys Asp Asn Ser Ile Asp Asn Lys 
    850                 855                 860                 


Val Leu Thr Ser Ser Lys Glu Asn Arg Gly Lys Ser Asp Asp Val Pro 
865                 870                 875                 880 


Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu 
                885                 890                 895     


Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala 
            900                 905                 910         


Glu Arg Gly Gly Leu Ser Glu Ala Asp Lys Ala Gly Phe Ile Lys Arg 
        915                 920                 925             


Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu 
    930                 935                 940                 


Asp Ser Arg Met Asn Thr Lys Arg Asp Lys Asn Asp Lys Pro Ile Arg 
945                 950                 955                 960 


Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg 
                965                 970                 975     


Lys Asp Phe Gln Leu Tyr Lys Val Arg Glu Ile Asn Asn Tyr His His 
            980                 985                 990         


Ala His Asp Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
        995                 1000                 1005             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1010                 1015                 1020             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1025                 1030                 1035             


Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1040                 1045                 1050             


Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1055                 1060                 1065             


Arg Pro  Leu Ile Glu Thr Asn  Glu Glu Thr Gly Glu  Ile Val Trp 
    1070                 1075                 1080             


Asp Lys  Glu Lys Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1085                 1090                 1095             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1100                 1105                 1110             


Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Gly Glu Ser  Asp Lys Leu 
    1115                 1120                 1125             


Ile Pro  Arg Lys Lys Gly Trp  Asp Ser Lys Lys Tyr  Gly Gly Phe 
    1130                 1135                 1140             


Gly Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1145                 1150                 1155             


Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Thr Val Lys  Glu Leu Val 
    1160                 1165                 1170             


Gly Ile  Thr Ile Met Glu Lys  Ser Ser Phe Glu Lys  Asp Pro Ile 
    1175                 1180                 1185             


Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp Ile Gln  Lys Asp Leu 
    1190                 1195                 1200             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1205                 1210                 1215             


Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn 
    1220                 1225                 1230             


Glu Leu  Val Leu Pro Gln His  Leu Val Thr Leu Leu  Tyr His Ala 
    1235                 1240                 1245             


His Lys  Ile Glu Ser Ser Lys  Glu Ser Glu Asn Gln  Ala Tyr Ile 
    1250                 1255                 1260             


Glu Gln  His Arg His Glu Phe  Asp Glu Ile Leu Glu  Tyr Ile Ile 
    1265                 1270                 1275             


Asp Phe  Ser Glu Lys Tyr Ile  Leu Ala Glu Lys Asn  Ile Ser Lys 
    1280                 1285                 1290             


Val Lys  Ser Leu Phe Asn Lys  Gln Phe Glu Asp Ser  Asp Ile Ser 
    1295                 1300                 1305             


Glu Leu  Ser Asn Ser Phe Ile  Asn Leu Leu Thr Leu  Thr Ser Phe 
    1310                 1315                 1320             


Gly Ala  Pro Ala Ala Phe Lys  Phe Leu Asp Thr Thr  Ile Lys Gln 
    1325                 1330                 1335             


Gly Arg  Lys Arg Tyr Gln Ser  Thr Lys Glu Val Leu  Asp Ala Thr 
    1340                 1345                 1350             


Leu Ile  His Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1355                 1360                 1365             


Leu Ser  Gln Leu Gly Gly Asp  
    1370                 1375 


<210>  269
<211>  986
<212>  PRT
<213>  Planctomycetes CasX


<220>
<221>  MISC_FEATURE
<223>  CasX

<400>  269

Met Glu Lys Arg Ile Asn Lys Ile Arg Lys Lys Leu Ser Ala Asp Asn 
1               5                   10                  15      


Ala Thr Lys Pro Val Ser Arg Ser Gly Pro Met Lys Thr Leu Leu Val 
            20                  25                  30          


Arg Val Met Thr Asp Asp Leu Lys Lys Arg Leu Glu Lys Arg Arg Lys 
        35                  40                  45              


Lys Pro Glu Val Met Pro Gln Val Ile Ser Asn Asn Ala Ala Asn Asn 
    50                  55                  60                  


Leu Arg Met Leu Leu Asp Asp Tyr Thr Lys Met Lys Glu Ala Ile Leu 
65                  70                  75                  80  


Gln Val Tyr Trp Gln Glu Phe Lys Asp Asp His Val Gly Leu Met Cys 
                85                  90                  95      


Lys Phe Ala Gln Pro Ala Ser Lys Lys Ile Asp Gln Asn Lys Leu Lys 
            100                 105                 110         


Pro Glu Met Asp Glu Lys Gly Asn Leu Thr Thr Ala Gly Phe Ala Cys 
        115                 120                 125             


Ser Gln Cys Gly Gln Pro Leu Phe Val Tyr Lys Leu Glu Gln Val Ser 
    130                 135                 140                 


Glu Lys Gly Lys Ala Tyr Thr Asn Tyr Phe Gly Arg Cys Asn Val Ala 
145                 150                 155                 160 


Glu His Glu Lys Leu Ile Leu Leu Ala Gln Leu Lys Pro Glu Lys Asp 
                165                 170                 175     


Ser Asp Glu Ala Val Thr Tyr Ser Leu Gly Lys Phe Gly Gln Arg Ala 
            180                 185                 190         


Leu Asp Phe Tyr Ser Ile His Val Thr Lys Glu Ser Thr His Pro Val 
        195                 200                 205             


Lys Pro Leu Ala Gln Ile Ala Gly Asn Arg Tyr Ala Ser Gly Pro Val 
    210                 215                 220                 


Gly Lys Ala Leu Ser Asp Ala Cys Met Gly Thr Ile Ala Ser Phe Leu 
225                 230                 235                 240 


Ser Lys Tyr Gln Asp Ile Ile Ile Glu His Gln Lys Val Val Lys Gly 
                245                 250                 255     


Asn Gln Lys Arg Leu Glu Ser Leu Arg Glu Leu Ala Gly Lys Glu Asn 
            260                 265                 270         


Leu Glu Tyr Pro Ser Val Thr Leu Pro Pro Gln Pro His Thr Lys Glu 
        275                 280                 285             


Gly Val Asp Ala Tyr Asn Glu Val Ile Ala Arg Val Arg Met Trp Val 
    290                 295                 300                 


Asn Leu Asn Leu Trp Gln Lys Leu Lys Leu Ser Arg Asp Asp Ala Lys 
305                 310                 315                 320 


Pro Leu Leu Arg Leu Lys Gly Phe Pro Ser Phe Pro Val Val Glu Arg 
                325                 330                 335     


Arg Glu Asn Glu Val Asp Trp Trp Asn Thr Ile Asn Glu Val Lys Lys 
            340                 345                 350         


Leu Ile Asp Ala Lys Arg Asp Met Gly Arg Val Phe Trp Ser Gly Val 
        355                 360                 365             


Thr Ala Glu Lys Arg Asn Thr Ile Leu Glu Gly Tyr Asn Tyr Leu Pro 
    370                 375                 380                 


Asn Glu Asn Asp His Lys Lys Arg Glu Gly Ser Leu Glu Asn Pro Lys 
385                 390                 395                 400 


Lys Pro Ala Lys Arg Gln Phe Gly Asp Leu Leu Leu Tyr Leu Glu Lys 
                405                 410                 415     


Lys Tyr Ala Gly Asp Trp Gly Lys Val Phe Asp Glu Ala Trp Glu Arg 
            420                 425                 430         


Ile Asp Lys Lys Ile Ala Gly Leu Thr Ser His Ile Glu Arg Glu Glu 
        435                 440                 445             


Ala Arg Asn Ala Glu Asp Ala Gln Ser Lys Ala Val Leu Thr Asp Trp 
    450                 455                 460                 


Leu Arg Ala Lys Ala Ser Phe Val Leu Glu Arg Leu Lys Glu Met Asp 
465                 470                 475                 480 


Glu Lys Glu Phe Tyr Ala Cys Glu Ile Gln Leu Gln Lys Trp Tyr Gly 
                485                 490                 495     


Asp Leu Arg Gly Asn Pro Phe Ala Val Glu Ala Glu Asn Arg Val Val 
            500                 505                 510         


Asp Ile Ser Gly Phe Ser Ile Gly Ser Asp Gly His Ser Ile Gln Tyr 
        515                 520                 525             


Arg Asn Leu Leu Ala Trp Lys Tyr Leu Glu Asn Gly Lys Arg Glu Phe 
    530                 535                 540                 


Tyr Leu Leu Met Asn Tyr Gly Lys Lys Gly Arg Ile Arg Phe Thr Asp 
545                 550                 555                 560 


Gly Thr Asp Ile Lys Lys Ser Gly Lys Trp Gln Gly Leu Leu Tyr Gly 
                565                 570                 575     


Gly Gly Lys Ala Lys Val Ile Asp Leu Thr Phe Asp Pro Asp Asp Glu 
            580                 585                 590         


Gln Leu Ile Ile Leu Pro Leu Ala Phe Gly Thr Arg Gln Gly Arg Glu 
        595                 600                 605             


Phe Ile Trp Asn Asp Leu Leu Ser Leu Glu Thr Gly Leu Ile Lys Leu 
    610                 615                 620                 


Ala Asn Gly Arg Val Ile Glu Lys Thr Ile Tyr Asn Lys Lys Ile Gly 
625                 630                 635                 640 


Arg Asp Glu Pro Ala Leu Phe Val Ala Leu Thr Phe Glu Arg Arg Glu 
                645                 650                 655     


Val Val Asp Pro Ser Asn Ile Lys Pro Val Asn Leu Ile Gly Val Asp 
            660                 665                 670         


Arg Gly Glu Asn Ile Pro Ala Val Ile Ala Leu Thr Asp Pro Glu Gly 
        675                 680                 685             


Cys Pro Leu Pro Glu Phe Lys Asp Ser Ser Gly Gly Pro Thr Asp Ile 
    690                 695                 700                 


Leu Arg Ile Gly Glu Gly Tyr Lys Glu Lys Gln Arg Ala Ile Gln Ala 
705                 710                 715                 720 


Ala Lys Glu Val Glu Gln Arg Arg Ala Gly Gly Tyr Ser Arg Lys Phe 
                725                 730                 735     


Ala Ser Lys Ser Arg Asn Leu Ala Asp Asp Met Val Arg Asn Ser Ala 
            740                 745                 750         


Arg Asp Leu Phe Tyr His Ala Val Thr His Asp Ala Val Leu Val Phe 
        755                 760                 765             


Glu Asn Leu Ser Arg Gly Phe Gly Arg Gln Gly Lys Arg Thr Phe Met 
    770                 775                 780                 


Thr Glu Arg Gln Tyr Thr Lys Met Glu Asp Trp Leu Thr Ala Lys Leu 
785                 790                 795                 800 


Ala Tyr Glu Gly Leu Thr Ser Lys Thr Tyr Leu Ser Lys Thr Leu Ala 
                805                 810                 815     


Gln Tyr Thr Ser Lys Thr Cys Ser Asn Cys Gly Phe Thr Ile Thr Thr 
            820                 825                 830         


Ala Asp Tyr Asp Gly Met Leu Val Arg Leu Lys Lys Thr Ser Asp Gly 
        835                 840                 845             


Trp Ala Thr Thr Leu Asn Asn Lys Glu Leu Lys Ala Glu Gly Gln Ile 
    850                 855                 860                 


Thr Tyr Tyr Asn Arg Tyr Lys Arg Gln Thr Val Glu Lys Glu Leu Ser 
865                 870                 875                 880 


Ala Glu Leu Asp Arg Leu Ser Glu Glu Ser Gly Asn Asn Asp Ile Ser 
                885                 890                 895     


Lys Trp Thr Lys Gly Arg Arg Asp Glu Ala Leu Phe Leu Leu Lys Lys 
            900                 905                 910         


Arg Phe Ser His Arg Pro Val Gln Glu Gln Phe Val Cys Leu Asp Cys 
        915                 920                 925             


Gly His Glu Val His Ala Asp Glu Gln Ala Ala Leu Asn Ile Ala Arg 
    930                 935                 940                 


Ser Trp Leu Phe Leu Asn Ser Asn Ser Thr Glu Phe Lys Ser Tyr Lys 
945                 950                 955                 960 


Ser Gly Lys Gln Pro Phe Val Gly Ala Trp Gln Ala Phe Tyr Lys Arg 
                965                 970                 975     


Arg Leu Lys Glu Val Trp Lys Pro Asn Ala 
            980                 985     


<210>  270
<211>  7959
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Plasmid construct 11235, expression dScCas9-FokI-NNG

<400>  270
ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc       60

attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga      120

gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt      180

gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt      240

gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg      300

acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca      360

aggccgcata tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg      420

cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt      480

gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca      540

atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc      600

aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta      660

catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac      720

catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg      780

atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg      840

ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt      900

acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta      960

ccggtatggc ccctaagaag aagagaaaag tcggcggctc tggcagccag ctggtcaagt     1020

ctgagctgga agagaagaaa agcgagctgc ggcacaagct gaaatacgtg ccccacgagt     1080

acatcgagct gatcgagatc gccagaaaca gcacccagga cagaatcctg gaaatgaagg     1140

tcatggaatt ctttatgaag gtgtacggct accggggcaa gcacctcgga ggatctagaa     1200

agcctgacgg cgccatctat accgtgggca gccctatcga ttacggcgtg atcgtggata     1260

ccaaggccta cagcggcggc tacaatctgc ctattggaca ggccgacgag atgcagagat     1320

acgtggaaga aaaccagaca cggaacaagc acatcaaccc caacgagtgg tggaaagtgt     1380

accccagcag cgtgaccgag ttcaagttcc tgttcgtgtc cggccacttc aagggcaact     1440

acaaggccca gctgacccgg ctgaaccaca tcacaaattg caacggcgct gtgctgtctg     1500

tggaagaact gctgatcggc ggcgagatga tcaaggccgg aacactgaca ctggaagaag     1560

tgcggcggaa gttcaacaac ggcgagatca acttcagcgg cagcgagaca cctggcacat     1620

ctgaatctgc cacacctgag agcatggaaa agaagtacag catcggcctg gccatcggca     1680

caaattctgt tggctgggcc gtgatcaccg acgactacaa ggtgcccagc aagaaattca     1740

aggtgctggg caacaccaac cggaagtcca tcaagaaaaa cctgatgggc gccctgctgt     1800

tcgattctgg cgaaacagcc gaggccacca gactgaagag aaccgccaga agaagataca     1860

cccggcggaa gaaccggatc agatacctgc aagagatctt cgccaacgag atggccaagc     1920

tggacgacag cttctttcag agactggaag agtccttcct ggtggaagag gacaagaaga     1980

acgagcggca ccccatcttc ggcaatctgg ctgatgaggt ggcctaccac agaaactacc     2040

ccaccatcta ccacctgagg aagaagctgg ccgactctcc tgagaaggcc gacctgagac     2100

tgatctatct ggccctggct cacatcatca agttccgggg ccacttcctg atcgagggca     2160

agctgaatgc cgagaacagc gacgtggcca aactgttcta ccagctgatc cagacctaca     2220

accagctgtt cgaggaaagc cctctggacg agattgaggt ggacgccaag ggaatcctga     2280

gcgccagact gagcaagagc aagcggctgg aaaagctgat cgccgtgttt cccaacgaga     2340

agaagaatgg cctgttcggc aacatcattg ccctggcact gggcctgaca cctaacttca     2400

agtccaactt cgacctgacc gaggacgcca aactgcagct gagcaaggac acctacgacg     2460

acgatctgga tgagctgctg ggccagatcg gagatcagta cgccgatctg ttcagcgccg     2520

ccaagaatct gtctgacgcc atcctgctga gcgacatcct gagatccaac agcgaagtga     2580

caaaggcccc tctgagcgcc tccatggtca agagatatga cgagcaccac caggacctgg     2640

ctctgctgaa aacccttgtg cggcagcagt tccccgagaa gtatgccgag atcttcaagg     2700

acgacaccaa gaacggctac gccggctatg ttggcatcgg catcaagcac agaaagcgga     2760

ccaccaagct ggccacacaa gaggaattct acaagttcat caagcccatc ctcgagaaga     2820

tggacggcgc cgaagaactc ctggctaagc tgaacaggga cgacctgctg cggaagcaga     2880

gaaccttcga caatggcagc atccctcacc agatccacct gaaagagctg cacgccattc     2940

tgcggcggca agaagagttc tacccattcc tgaaagaaaa ccgggaaaag attgagaaga     3000

tcctgacctt caggatcccc tactacgtgg gaccactggc cagaggcaat agcagattcg     3060

cttggctgac cagaaagagc gaggaagcca tcacaccctg gaacttcgag gaagtggtgg     3120

ataagggcgc cagcgctcag tccttcatcg agcggatgac caatttcgac gagcagctgc     3180

ccaacaagaa ggtgctgcct aagcacagcc tgctgtacga gtacttcacc gtgtacaatg     3240

agctgaccaa agtcaaatac gtgacagagc ggatgcggaa gcctgagttt ctgagcggcg     3300

agcagaaaaa ggccatcgtg gatctgctgt tcaagaccaa cagaaaagtg accgtgaagc     3360

agctgaaaga ggactacttc aagaaaatcg agtgcttcga cagcgtcgag atcatcggcg     3420

tggaagatag attcaacgcc agcctgggca cataccacga tctgctgaag attatcaagg     3480

acaaggactt cctggacaac gaggaaaacg aggacatcct ggaagatatc gtgctgaccc     3540

tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacatac gcccacctgt     3600

tcgacgacaa agtgatgaag caactgaagc ggcggcacta cacaggctgg ggcagactgt     3660

ccagaaagat gattaacggc atccgggaca agcagagcgg caagaccatc ctggactttc     3720

tgaagtccga cggcttcagc aaccggaact tcatgcagct gattcacgac gacagcctca     3780

ccttcaaaga ggaaatcgag aaggcccagg tttccggcca gggcgattct ctgcacgagc     3840

agattgctga tctggccgga tctcccgcca ttaagaaggg catcctgcag acagtgaaga     3900

tcgtggacga actcgtgaaa gtgatgggcc acaagcctga gaacatcgtg atcgaaatgg     3960

ccagagagaa ccagaccacc accaagggcc tgcagcagtc cagagagaga aagaagcgga     4020

tcgaagaggg catcaaagag ctggaaagcc agatcctcaa agagaacccc gtcgagaaca     4080

cccagctgca gaacgagaag ctgtacctgt actacctgca gaatggacgg gatatgtacg     4140

tggaccaaga actggacatc aaccggctgt ccgactacga tgtggatgcc atcgtgcccc     4200

agagcttcat caaggatgac agcatcgaca acaaggtcct gaccagatcc gtggaaaaca     4260

gaggcaagag cgacaacgtg ccctccgaag aggtggtcaa gaagatgaag aattactggc     4320

gacagctgct gaacgccaag ctgatcaccc agagaaagtt cgacaacctg accaaggccg     4380

agagaggcgg actgtctgaa gctgataagg ccggcttcat taagcggcag ctggtggaaa     4440

cccggcagat cacaaagcac gtggcccgga tcctggacag cagaatgaac accaagcggg     4500

acaagaacga caagcctatc cgggaagtga aagtcatcac cctgaagtcc aagctggtgt     4560

ccgatttccg gaaggatttc cagctgtaca aagtgcgcga catcaacaac taccatcacg     4620

cccacgacgc ctacctgaat gctgttgtgg gaacagccct gatcaagaag taccctaagc     4680

tggaatccga gttcgtgtac ggcgactaca aagtgtacga cgtgcggaag atgatcgcca     4740

agagcgagca agagattggc aaggccaccg ccaagcggtt cttctactcc aacatcatga     4800

actttttcaa gaccgaagtg aagctcgcca acggcgaaat ccggaagagg cccctgattg     4860

agacaaatgg cgagacaggc gaggtcgtgt ggaacaaaga gaaggacttc gccaccgtgc     4920

ggaaagtgct ggccatgcct caagtgaata tcgtgaaaaa gaccgaggtg cagaccggcg     4980

gcttctccaa agagagcatc ctgtctaaga gagagagcgc caagctcatc cctcggaaga     5040

aaggctggga tacacggaag tacggcggat ttggaagccc taccgtggcc tactctattc     5100

tggtggtggc caaggtggaa aagggcaaag ccaagaaact gaagtctgtg aaggtgctcg     5160

tggggatcac catcatggaa aaaggcagct acgagaagga ccctatcggc tttctggaag     5220

ccaagggcta caaggacatc aaaaaagagc tgatctttaa gctccccaag tacagcctgt     5280

tcgaactgga aaacggcaga cgcagaatgc tggcctctgc cacagaactg cagaaagcca     5340

atgaactggt gctgccacaa cacctcgtgc ggctgctgta ctacacccag aatatcagcg     5400

ccaccaccgg cagcaacaac ctgggatata tcgagcagca ccgggaagag ttcaaagaga     5460

ttttcgagaa aatcatcgac ttcagcgaga agtacatcct gaagaacaaa gtgaacagca     5520

acctcaagag cagcttcgat gagcagttcg ccgtgtccga ctctatcctg ctgtctaaca     5580

gcttcgtgtc cctgctgaag tacacctcct ttggcgcctc tggcggcttc acctttctgg     5640

atctggacgt gaaacagggg agactgagat accagaccgt gactgaggtg ctggacgcca     5700

ctctgatcta ccagagcatc accggcctgt acgagacaag aaccgatctc tctcagctcg     5760

gaggcgacta aaagcttctg tgccttctag ttgccagcca tctgttgttt gcccctcccc     5820

cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga     5880

aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga     5940

cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat     6000

ggacaaaccc acaactacgc gtacttgcgg ccgcatattc ctaggatatt ctgggcctca     6060

tgggccttcc tttcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa     6120

catggtcata gctgtttcct tgcgtattgg gcgctctccg cttcctcgct cactgactcg     6180

ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc cagcaaaagg     6240

ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg     6300

agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat     6360

accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta     6420

ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct     6480

gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc     6540

ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa     6600

gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg     6660

taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag     6720

tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt     6780

gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta     6840

cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc     6900

agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca     6960

cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa     7020

cttggtctga cagttattag aaaaattcat ccagcagacg ataaaacgca atacgctggc     7080

tatccggtgc cgcaatgcca tacagcacca gaaaacgatc cgcccattcg ccgcccagtt     7140

cttccgcaat atcacgggtg gccagcgcaa tatcctgata acgatccgcc acgcccagac     7200

ggccgcaatc aataaagccg ctaaaacggc cattttccac cataatgttc ggcaggcacg     7260

catcaccatg ggtcaccacc agatcttcgc catccggcat gctcgctttc agacgcgcaa     7320

acagctctgc cggtgccagg ccctgatgtt cttcatccag atcatcctga tccaccaggc     7380

ccgcttccat acgggtacgc gcacgttcaa tacgatgttt cgcctgatga tcaaacggac     7440

aggtcgccgg gtccagggta tgcagacgac gcatggcatc cgccataatg ctcacttttt     7500

ctgccggcgc cagatggcta gacagcagat cctgacccgg cacttcgccc agcagcagcc     7560

aatcacggcc cgcttcggtc accacatcca gcaccgccgc acacggaaca ccggtggtgg     7620

ccagccagct cagacgcgcc gcttcatcct gcagctcgtt cagcgcaccg ctcagatcgg     7680

ttttcacaaa cagcaccgga cgaccctgcg cgctcagacg aaacaccgcc gcatcagagc     7740

agccaatggt ctgctgcgcc caatcatagc caaacagacg ttccacccac gctgccgggc     7800

tacccgcatg caggccatcc tgttcaatca tactcttcct ttttcaatat tattgaagca     7860

tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac     7920

aaataggggt tccgcgcaca tttccccgaa aagtgccac                            7959


<210>  271
<211>  760
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCas14-FokI

<400>  271

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
225                 230                 235                 240 


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
                245                 250                 255     


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
            260                 265                 270         


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
        275                 280                 285             


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
    290                 295                 300                 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
305                 310                 315                 320 


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
                325                 330                 335     


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
            340                 345                 350         


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
        355                 360                 365             


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
    370                 375                 380                 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
385                 390                 395                 400 


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
                405                 410                 415     


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
            420                 425                 430         


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
        435                 440                 445             


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
    450                 455                 460                 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
465                 470                 475                 480 


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
                485                 490                 495     


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
            500                 505                 510         


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
        515                 520                 525             


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
    530                 535                 540                 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
545                 550                 555                 560 


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
                565                 570                 575     


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
            580                 585                 590         


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
        595                 600                 605             


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
    610                 615                 620                 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
625                 630                 635                 640 


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
                645                 650                 655     


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
            660                 665                 670         


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
        675                 680                 685             


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
    690                 695                 700                 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
705                 710                 715                 720 


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
                725                 730                 735     


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
            740                 745                 750         


Leu Lys Ser Thr Lys Glu Glu Pro 
        755                 760 


<210>  272
<211>  818
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  StkC-dCas14-FokI

<400>  272

Met Ala Val Thr Ser Val Ser Met Val His Ser Glu Arg Leu Pro Leu 
1               5                   10                  15      


Leu Met Asn Arg Asn Gly Ile Arg Phe Gln Ser Glu Val Ile Asp Pro 
            20                  25                  30          


Asn Pro Pro Lys Lys Val Lys Trp Val Leu Thr Gly Ser Gly Gly Asn 
        35                  40                  45              


Met Phe Thr Ala Asn Val Arg Gly Ser Asn Val Ser Ser Leu Val Lys 
    50                  55                  60                  


His Thr Trp Ser Glu Ala Phe Val Ser Leu Ile Glu Asp Arg Val Lys 
65                  70                  75                  80  


Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr 
                85                  90                  95      


Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr 
            100                 105                 110         


Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met Lys Val 
        115                 120                 125             


Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro Asp Gly 
    130                 135                 140                 


Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile Val Asp 
145                 150                 155                 160 


Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala Asp 
                165                 170                 175     


Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile 
            180                 185                 190         


Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe 
        195                 200                 205             


Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys Ala Gln 
    210                 215                 220                 


Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val Leu Ser 
225                 230                 235                 240 


Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly Thr Leu 
                245                 250                 255     


Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile Asn Phe 
            260                 265                 270         


Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Gly 
        275                 280                 285             


Thr Met Ala Lys Asn Thr Ile Thr Lys Thr Leu Lys Leu Arg Ile Val 
    290                 295                 300                 


Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys Ile Val Ala Asp Glu Lys 
305                 310                 315                 320 


Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys Asn Lys Asp Lys Val Lys 
                325                 330                 335     


Glu Ala Cys Ser Lys His Leu Lys Val Ala Ala Tyr Cys Thr Thr Gln 
            340                 345                 350         


Val Glu Arg Asn Ala Cys Leu Phe Cys Lys Ala Arg Lys Leu Asp Asp 
        355                 360                 365             


Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe Pro Asp Ala Val Phe Trp 
    370                 375                 380                 


Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu Gln Lys Gln Ala Ala Glu 
385                 390                 395                 400 


Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr Tyr Glu Ile Phe Ile Lys 
                405                 410                 415     


Gly Lys Gly Ile Ala Asn Ala Ser Ser Val Glu His Tyr Leu Ser Asp 
            420                 425                 430         


Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe Lys Asn Ala Ala Ile Ala 
        435                 440                 445             


Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn Phe Arg Leu Lys Glu Leu 
    450                 455                 460                 


Lys Asn Met Lys Ser Gly Leu Pro Thr Thr Lys Ser Asp Asn Phe Pro 
465                 470                 475                 480 


Ile Pro Leu Val Lys Gln Lys Gly Gly Gln Tyr Thr Gly Phe Glu Ile 
                485                 490                 495     


Ser Asn His Asn Ser Asp Phe Ile Ile Lys Ile Pro Phe Gly Arg Trp 
            500                 505                 510         


Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg Pro Trp Glu Lys Phe Asp 
        515                 520                 525             


Phe Glu Gln Val Gln Lys Ser Pro Lys Pro Ile Ser Leu Leu Leu Ser 
    530                 535                 540                 


Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp Ser Lys Asp Glu Gly Thr 
545                 550                 555                 560 


Glu Ala Glu Ile Lys Lys Val Met Asn Gly Asp Tyr Gln Thr Ser Tyr 
                565                 570                 575     


Ile Glu Val Lys Arg Gly Ser Lys Ile Cys Glu Lys Ser Ala Trp Met 
            580                 585                 590         


Leu Asn Leu Ser Ile Asp Val Pro Lys Ile Asp Lys Gly Val Asp Pro 
        595                 600                 605             


Ser Ile Ile Gly Gly Ile Ala Val Gly Val Lys Ser Pro Leu Val Cys 
    610                 615                 620                 


Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser Ile Ser Asp Asn Asp Leu 
625                 630                 635                 640 


Phe His Phe Asn Lys Lys Met Phe Ala Arg Arg Arg Ile Leu Leu Lys 
                645                 650                 655     


Lys Asn Arg His Lys Arg Ala Gly His Gly Ala Lys Asn Lys Leu Lys 
            660                 665                 670         


Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu Arg Phe Arg Lys Lys Leu 
        675                 680                 685             


Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp Phe Phe Ile Lys Asn Lys 
    690                 695                 700                 


Val Gly Thr Val Gln Met Glu Asn Leu Glu Ser Met Lys Arg Lys Glu 
705                 710                 715                 720 


Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly Phe Trp Pro Tyr Ala Glu 
                725                 730                 735     


Met Gln Asn Lys Ile Glu Phe Lys Leu Lys Gln Tyr Gly Ile Glu Ile 
            740                 745                 750         


Arg Lys Val Ala Pro Asn Asn Thr Ser Lys Thr Cys Ser Lys Cys Gly 
        755                 760                 765             


His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr Arg Lys Lys Asn Lys Phe 
    770                 775                 780                 


Pro His Phe Lys Cys Glu Lys Cys Asn Phe Lys Glu Asn Ala Asp Tyr 
785                 790                 795                 800 


Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys Leu Lys Ser Thr Lys Glu 
                805                 810                 815     


Glu Pro 
        


<210>  273
<211>  795
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HIVINT-dCas14-FokI

<400>  273

Met Ala Val Thr Ile Gln Asn Phe Arg Val Tyr Tyr Arg Asp Ser Arg 
1               5                   10                  15      


Asn Pro Leu Trp Lys Gly Pro Ala Lys Leu Leu Trp Lys Gly Glu Gly 
            20                  25                  30          


Ala Trp Ile Gln Asp Asn Ser Asp Ile Lys Trp Pro Arg Arg Lys Ala 
        35                  40                  45              


Lys Ile Ile Arg Asp Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu 
    50                  55                  60                  


Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile 
65                  70                  75                  80  


Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val 
                85                  90                  95      


Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly 
            100                 105                 110         


Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile 
        115                 120                 125             


Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn 
    130                 135                 140                 


Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn 
145                 150                 155                 160 


Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr 
                165                 170                 175     


Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe 
            180                 185                 190         


Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn 
        195                 200                 205             


Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu 
    210                 215                 220                 


Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe 
225                 230                 235                 240 


Asn Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Lys Lys Lys 
                245                 250                 255     


Arg Lys Val Pro Gly Thr Ser Glu Gly Thr Met Ala Lys Asn Thr Ile 
            260                 265                 270         


Thr Lys Thr Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu 
        275                 280                 285             


Val Glu Lys Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala 
    290                 295                 300                 


Leu Glu Lys Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu 
305                 310                 315                 320 


Lys Val Ala Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu 
                325                 330                 335     


Phe Cys Lys Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg 
            340                 345                 350         


Gly Gln Phe Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe 
        355                 360                 365             


Arg Gln Leu Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile 
    370                 375                 380                 


Glu Leu Tyr Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala 
385                 390                 395                 400 


Ser Ser Val Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala 
                405                 410                 415     


Glu Leu Phe Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile 
            420                 425                 430         


Lys Ser Asn Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu 
        435                 440                 445             


Pro Thr Thr Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys 
    450                 455                 460                 


Gly Gly Gln Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe 
465                 470                 475                 480 


Ile Ile Lys Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp 
                485                 490                 495     


Lys Tyr Arg Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser 
            500                 505                 510         


Pro Lys Pro Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn 
        515                 520                 525             


Lys Gly Trp Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val 
    530                 535                 540                 


Met Asn Gly Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser 
545                 550                 555                 560 


Lys Ile Cys Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val 
                565                 570                 575     


Pro Lys Ile Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala 
            580                 585                 590         


Val Gly Val Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser 
        595                 600                 605             


Arg Tyr Ser Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met 
    610                 615                 620                 


Phe Ala Arg Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala 
625                 630                 635                 640 


Gly His Gly Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu 
                645                 650                 655     


Lys Ser Glu Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu 
            660                 665                 670         


Ile Ala Asp Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu 
        675                 680                 685             


Asn Leu Glu Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg 
    690                 695                 700                 


Leu Arg Gly Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe 
705                 710                 715                 720 


Lys Leu Lys Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn 
                725                 730                 735     


Thr Ser Lys Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn 
            740                 745                 750         


Phe Glu Tyr Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys 
        755                 760                 765             


Cys Asn Phe Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser 
    770                 775                 780                 


Asn Pro Lys Leu Lys Ser Thr Lys Glu Glu Pro 
785                 790                 795 


<210>  274
<211>  810
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SS07D-dCas14-FokI

<400>  274

Met Ala Val Thr Met Ala Thr Val Lys Phe Lys Tyr Lys Gly Glu Glu 
1               5                   10                  15      


Lys Glu Val Asp Ile Ser Lys Ile Lys Lys Val Trp Arg Val Gly Lys 
            20                  25                  30          


Met Ile Ser Phe Thr Tyr Asp Glu Gly Gly Gly Lys Thr Gly Arg Gly 
        35                  40                  45              


Ala Val Ser Glu Lys Asp Ala Pro Lys Glu Leu Leu Gln Met Leu Glu 
    50                  55                  60                  


Lys Gln Lys Lys Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu 
65                  70                  75                  80  


Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu 
                85                  90                  95      


Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met 
            100                 105                 110         


Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly 
        115                 120                 125             


Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp 
    130                 135                 140                 


Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu 
145                 150                 155                 160 


Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln 
                165                 170                 175     


Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro 
            180                 185                 190         


Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys 
        195                 200                 205             


Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys 
    210                 215                 220                 


Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met 
225                 230                 235                 240 


Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn 
                245                 250                 255     


Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Lys Lys Lys Arg 
            260                 265                 270         


Lys Val Pro Gly Thr Ser Glu Gly Thr Met Ala Lys Asn Thr Ile Thr 
        275                 280                 285             


Lys Thr Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val 
    290                 295                 300                 


Glu Lys Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu 
305                 310                 315                 320 


Glu Lys Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys 
                325                 330                 335     


Val Ala Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe 
            340                 345                 350         


Cys Lys Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly 
        355                 360                 365             


Gln Phe Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg 
    370                 375                 380                 


Gln Leu Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu 
385                 390                 395                 400 


Leu Tyr Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser 
                405                 410                 415     


Ser Val Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu 
            420                 425                 430         


Leu Phe Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys 
        435                 440                 445             


Ser Asn Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro 
    450                 455                 460                 


Thr Thr Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly 
465                 470                 475                 480 


Gly Gln Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile 
                485                 490                 495     


Ile Lys Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys 
            500                 505                 510         


Tyr Arg Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro 
        515                 520                 525             


Lys Pro Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys 
    530                 535                 540                 


Gly Trp Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met 
545                 550                 555                 560 


Asn Gly Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys 
                565                 570                 575     


Ile Cys Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro 
            580                 585                 590         


Lys Ile Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val 
        595                 600                 605             


Gly Val Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg 
    610                 615                 620                 


Tyr Ser Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe 
625                 630                 635                 640 


Ala Arg Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly 
                645                 650                 655     


His Gly Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys 
            660                 665                 670         


Ser Glu Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile 
        675                 680                 685             


Ala Asp Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn 
    690                 695                 700                 


Leu Glu Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu 
705                 710                 715                 720 


Arg Gly Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys 
                725                 730                 735     


Leu Lys Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr 
            740                 745                 750         


Ser Lys Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe 
        755                 760                 765             


Glu Tyr Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys 
    770                 775                 780                 


Asn Phe Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn 
785                 790                 795                 800 


Pro Lys Leu Lys Ser Thr Lys Glu Glu Pro 
                805                 810 


<210>  275
<211>  856
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dCas14-FokI

<400>  275

Met Gly Pro Lys Lys Lys Arg Lys Val Met Pro Lys Arg Lys Val Ser 
1               5                   10                  15      


Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg 
            20                  25                  30          


Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala 
        35                  40                  45              


Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys 
    50                  55                  60                  


Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys 
65                  70                  75                  80  


Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro 
                85                  90                  95      


Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Pro Lys Lys 
            100                 105                 110         


Lys Arg Lys Val Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu 
        115                 120                 125             


Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu 
    130                 135                 140                 


Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met 
145                 150                 155                 160 


Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly 
                165                 170                 175     


Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp 
            180                 185                 190         


Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu 
        195                 200                 205             


Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln 
    210                 215                 220                 


Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro 
225                 230                 235                 240 


Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys 
                245                 250                 255     


Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys 
            260                 265                 270         


Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met 
        275                 280                 285             


Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn 
    290                 295                 300                 


Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu 
305                 310                 315                 320 


Ser Ala Thr Pro Glu Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
                325                 330                 335     


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
            340                 345                 350         


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
        355                 360                 365             


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
    370                 375                 380                 


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
385                 390                 395                 400 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
                405                 410                 415     


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
            420                 425                 430         


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
        435                 440                 445             


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
    450                 455                 460                 


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
465                 470                 475                 480 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
                485                 490                 495     


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
            500                 505                 510         


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
        515                 520                 525             


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
    530                 535                 540                 


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
545                 550                 555                 560 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
                565                 570                 575     


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
            580                 585                 590         


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
        595                 600                 605             


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
    610                 615                 620                 


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
625                 630                 635                 640 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
                645                 650                 655     


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
            660                 665                 670         


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
        675                 680                 685             


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
    690                 695                 700                 


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
705                 710                 715                 720 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
                725                 730                 735     


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
            740                 745                 750         


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
        755                 760                 765             


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
    770                 775                 780                 


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
785                 790                 795                 800 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
                805                 810                 815     


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
            820                 825                 830         


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
        835                 840                 845             


Leu Lys Ser Thr Lys Glu Glu Pro 
    850                 855     


<210>  276
<211>  939
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dCas14-FokI-HMGB1

<400>  276

Met Gly Pro Lys Lys Lys Arg Lys Val Met Pro Lys Arg Lys Val Ser 
1               5                   10                  15      


Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg 
            20                  25                  30          


Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala 
        35                  40                  45              


Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys 
    50                  55                  60                  


Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys 
65                  70                  75                  80  


Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro 
                85                  90                  95      


Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Pro Lys Lys 
            100                 105                 110         


Lys Arg Lys Val Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu 
        115                 120                 125             


Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu 
    130                 135                 140                 


Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met 
145                 150                 155                 160 


Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly 
                165                 170                 175     


Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp 
            180                 185                 190         


Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu 
        195                 200                 205             


Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln 
    210                 215                 220                 


Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro 
225                 230                 235                 240 


Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys 
                245                 250                 255     


Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys 
            260                 265                 270         


Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met 
        275                 280                 285             


Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn 
    290                 295                 300                 


Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu 
305                 310                 315                 320 


Ser Ala Thr Pro Glu Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
                325                 330                 335     


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
            340                 345                 350         


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
        355                 360                 365             


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
    370                 375                 380                 


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
385                 390                 395                 400 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
                405                 410                 415     


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
            420                 425                 430         


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
        435                 440                 445             


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
    450                 455                 460                 


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
465                 470                 475                 480 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
                485                 490                 495     


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
            500                 505                 510         


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
        515                 520                 525             


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
    530                 535                 540                 


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
545                 550                 555                 560 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
                565                 570                 575     


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
            580                 585                 590         


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
        595                 600                 605             


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
    610                 615                 620                 


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
625                 630                 635                 640 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
                645                 650                 655     


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
            660                 665                 670         


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
        675                 680                 685             


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
    690                 695                 700                 


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
705                 710                 715                 720 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
                725                 730                 735     


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
            740                 745                 750         


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
        755                 760                 765             


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
    770                 775                 780                 


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
785                 790                 795                 800 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
                805                 810                 815     


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
            820                 825                 830         


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
        835                 840                 845             


Leu Lys Ser Thr Lys Glu Glu Pro Gly Lys Gly Asp Pro Lys Lys Pro 
    850                 855                 860                 


Arg Gly Lys Met Ser Ser Tyr Ala Phe Phe Val Gln Thr Cys Arg Glu 
865                 870                 875                 880 


Glu His Lys Lys Lys His Pro Asp Ala Ser Val Asn Phe Ser Glu Phe 
                885                 890                 895     


Ser Lys Lys Cys Ser Glu Arg Trp Lys Thr Met Ser Ala Lys Glu Lys 
            900                 905                 910         


Gly Lys Phe Glu Asp Met Ala Lys Ala Asp Lys Ala Arg Tyr Glu Arg 
        915                 920                 925             


Glu Met Lys Thr Tyr Ile Pro Pro Lys Gly Glu 
    930                 935                 


<210>  277
<211>  942
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dCas14-FokI-HMGB3

<400>  277

Met Gly Pro Lys Lys Lys Arg Lys Val Met Pro Lys Arg Lys Val Ser 
1               5                   10                  15      


Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg 
            20                  25                  30          


Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala 
        35                  40                  45              


Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys 
    50                  55                  60                  


Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys 
65                  70                  75                  80  


Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro 
                85                  90                  95      


Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Pro Lys Lys 
            100                 105                 110         


Lys Arg Lys Val Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu 
        115                 120                 125             


Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu 
    130                 135                 140                 


Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met 
145                 150                 155                 160 


Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly 
                165                 170                 175     


Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp 
            180                 185                 190         


Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu 
        195                 200                 205             


Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln 
    210                 215                 220                 


Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro 
225                 230                 235                 240 


Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys 
                245                 250                 255     


Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys 
            260                 265                 270         


Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met 
        275                 280                 285             


Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn 
    290                 295                 300                 


Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu 
305                 310                 315                 320 


Ser Ala Thr Pro Glu Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
                325                 330                 335     


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
            340                 345                 350         


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
        355                 360                 365             


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
    370                 375                 380                 


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
385                 390                 395                 400 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
                405                 410                 415     


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
            420                 425                 430         


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
        435                 440                 445             


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
    450                 455                 460                 


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
465                 470                 475                 480 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
                485                 490                 495     


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
            500                 505                 510         


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
        515                 520                 525             


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
    530                 535                 540                 


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
545                 550                 555                 560 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
                565                 570                 575     


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
            580                 585                 590         


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
        595                 600                 605             


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
    610                 615                 620                 


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
625                 630                 635                 640 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
                645                 650                 655     


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
            660                 665                 670         


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
        675                 680                 685             


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
    690                 695                 700                 


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
705                 710                 715                 720 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
                725                 730                 735     


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
            740                 745                 750         


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
        755                 760                 765             


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
    770                 775                 780                 


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
785                 790                 795                 800 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
                805                 810                 815     


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
            820                 825                 830         


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
        835                 840                 845             


Leu Lys Ser Thr Lys Glu Glu Pro Gly Ser Ser Gly Ser Ser Gly Met 
    850                 855                 860                 


Ala Lys Gly Asp Pro Lys Lys Pro Lys Gly Lys Met Ser Ala Tyr Ala 
865                 870                 875                 880 


Phe Phe Val Gln Thr Cys Arg Glu Glu His Lys Lys Lys Asn Pro Glu 
                885                 890                 895     


Val Pro Val Asn Phe Ala Glu Phe Ser Lys Lys Cys Ser Glu Arg Trp 
            900                 905                 910         


Lys Thr Met Ser Gly Lys Glu Lys Ser Lys Phe Asp Glu Met Ala Lys 
        915                 920                 925             


Ala Asp Lys Val Arg Tyr Asp Arg Glu Met Lys Asp Tyr Gly 
    930                 935                 940         


<210>  278
<211>  934
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dCas14-FokI-HMGB4

<400>  278

Met Gly Pro Lys Lys Lys Arg Lys Val Met Pro Lys Arg Lys Val Ser 
1               5                   10                  15      


Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg 
            20                  25                  30          


Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala 
        35                  40                  45              


Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys 
    50                  55                  60                  


Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys 
65                  70                  75                  80  


Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro 
                85                  90                  95      


Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Pro Lys Lys 
            100                 105                 110         


Lys Arg Lys Val Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu 
        115                 120                 125             


Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu 
    130                 135                 140                 


Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met 
145                 150                 155                 160 


Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly 
                165                 170                 175     


Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp 
            180                 185                 190         


Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu 
        195                 200                 205             


Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln 
    210                 215                 220                 


Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro 
225                 230                 235                 240 


Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys 
                245                 250                 255     


Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys 
            260                 265                 270         


Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met 
        275                 280                 285             


Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn 
    290                 295                 300                 


Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu 
305                 310                 315                 320 


Ser Ala Thr Pro Glu Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
                325                 330                 335     


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
            340                 345                 350         


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
        355                 360                 365             


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
    370                 375                 380                 


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
385                 390                 395                 400 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
                405                 410                 415     


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
            420                 425                 430         


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
        435                 440                 445             


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
    450                 455                 460                 


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
465                 470                 475                 480 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
                485                 490                 495     


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
            500                 505                 510         


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
        515                 520                 525             


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
    530                 535                 540                 


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
545                 550                 555                 560 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
                565                 570                 575     


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
            580                 585                 590         


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
        595                 600                 605             


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
    610                 615                 620                 


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
625                 630                 635                 640 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
                645                 650                 655     


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
            660                 665                 670         


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
        675                 680                 685             


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
    690                 695                 700                 


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
705                 710                 715                 720 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
                725                 730                 735     


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
            740                 745                 750         


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
        755                 760                 765             


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
    770                 775                 780                 


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
785                 790                 795                 800 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
                805                 810                 815     


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
            820                 825                 830         


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
        835                 840                 845             


Leu Lys Ser Thr Lys Glu Glu Pro Gly Lys Glu Ile Gln Leu Lys Pro 
    850                 855                 860                 


Lys Ala Asn Val Ser Ser Tyr Val His Phe Leu Leu Asn Tyr Arg Asn 
865                 870                 875                 880 


Lys Phe Lys Glu Gln Gln Pro Asn Thr Tyr Val Gly Phe Lys Glu Phe 
                885                 890                 895     


Ser Arg Lys Cys Ser Glu Lys Trp Arg Ser Ile Ser Lys His Glu Lys 
            900                 905                 910         


Ala Lys Tyr Glu Ala Leu Ala Lys Leu Asp Lys Ala Arg Tyr Gln Glu 
        915                 920                 925             


Glu Met Met Asn Tyr Val 
    930                 


<210>  279
<211>  843
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCas14-FokI-HMGB1

<400>  279

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
225                 230                 235                 240 


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
                245                 250                 255     


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
            260                 265                 270         


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
        275                 280                 285             


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
    290                 295                 300                 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
305                 310                 315                 320 


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
                325                 330                 335     


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
            340                 345                 350         


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
        355                 360                 365             


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
    370                 375                 380                 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
385                 390                 395                 400 


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
                405                 410                 415     


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
            420                 425                 430         


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
        435                 440                 445             


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
    450                 455                 460                 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
465                 470                 475                 480 


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
                485                 490                 495     


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
            500                 505                 510         


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
        515                 520                 525             


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
    530                 535                 540                 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
545                 550                 555                 560 


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
                565                 570                 575     


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
            580                 585                 590         


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
        595                 600                 605             


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
    610                 615                 620                 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
625                 630                 635                 640 


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
                645                 650                 655     


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
            660                 665                 670         


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
        675                 680                 685             


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
    690                 695                 700                 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
705                 710                 715                 720 


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
                725                 730                 735     


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
            740                 745                 750         


Leu Lys Ser Thr Lys Glu Glu Pro Gly Lys Gly Asp Pro Lys Lys Pro 
        755                 760                 765             


Arg Gly Lys Met Ser Ser Tyr Ala Phe Phe Val Gln Thr Cys Arg Glu 
    770                 775                 780                 


Glu His Lys Lys Lys His Pro Asp Ala Ser Val Asn Phe Ser Glu Phe 
785                 790                 795                 800 


Ser Lys Lys Cys Ser Glu Arg Trp Lys Thr Met Ser Ala Lys Glu Lys 
                805                 810                 815     


Gly Lys Phe Glu Asp Met Ala Lys Ala Asp Lys Ala Arg Tyr Glu Arg 
            820                 825                 830         


Glu Met Lys Thr Tyr Ile Pro Pro Lys Gly Glu 
        835                 840             


<210>  280
<211>  846
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCas14-FokI-HMGB3

<400>  280

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
225                 230                 235                 240 


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
                245                 250                 255     


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
            260                 265                 270         


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
        275                 280                 285             


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
    290                 295                 300                 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
305                 310                 315                 320 


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
                325                 330                 335     


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
            340                 345                 350         


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
        355                 360                 365             


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
    370                 375                 380                 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
385                 390                 395                 400 


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
                405                 410                 415     


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
            420                 425                 430         


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
        435                 440                 445             


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
    450                 455                 460                 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
465                 470                 475                 480 


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
                485                 490                 495     


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
            500                 505                 510         


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
        515                 520                 525             


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
    530                 535                 540                 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
545                 550                 555                 560 


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
                565                 570                 575     


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
            580                 585                 590         


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
        595                 600                 605             


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
    610                 615                 620                 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
625                 630                 635                 640 


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
                645                 650                 655     


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
            660                 665                 670         


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
        675                 680                 685             


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
    690                 695                 700                 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
705                 710                 715                 720 


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
                725                 730                 735     


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
            740                 745                 750         


Leu Lys Ser Thr Lys Glu Glu Pro Gly Ser Ser Gly Ser Ser Gly Met 
        755                 760                 765             


Ala Lys Gly Asp Pro Lys Lys Pro Lys Gly Lys Met Ser Ala Tyr Ala 
    770                 775                 780                 


Phe Phe Val Gln Thr Cys Arg Glu Glu His Lys Lys Lys Asn Pro Glu 
785                 790                 795                 800 


Val Pro Val Asn Phe Ala Glu Phe Ser Lys Lys Cys Ser Glu Arg Trp 
                805                 810                 815     


Lys Thr Met Ser Gly Lys Glu Lys Ser Lys Phe Asp Glu Met Ala Lys 
            820                 825                 830         


Ala Asp Lys Val Arg Tyr Asp Arg Glu Met Lys Asp Tyr Gly 
        835                 840                 845     


<210>  281
<211>  838
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCas14-FokI-HMGB4

<400>  281

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Met Ala Lys Asn Thr Ile Thr Lys Thr 
225                 230                 235                 240 


Leu Lys Leu Arg Ile Val Arg Pro Tyr Asn Ser Ala Glu Val Glu Lys 
                245                 250                 255     


Ile Val Ala Asp Glu Lys Asn Asn Arg Glu Lys Ile Ala Leu Glu Lys 
            260                 265                 270         


Asn Lys Asp Lys Val Lys Glu Ala Cys Ser Lys His Leu Lys Val Ala 
        275                 280                 285             


Ala Tyr Cys Thr Thr Gln Val Glu Arg Asn Ala Cys Leu Phe Cys Lys 
    290                 295                 300                 


Ala Arg Lys Leu Asp Asp Lys Phe Tyr Gln Lys Leu Arg Gly Gln Phe 
305                 310                 315                 320 


Pro Asp Ala Val Phe Trp Gln Glu Ile Ser Glu Ile Phe Arg Gln Leu 
                325                 330                 335     


Gln Lys Gln Ala Ala Glu Ile Tyr Asn Gln Ser Leu Ile Glu Leu Tyr 
            340                 345                 350         


Tyr Glu Ile Phe Ile Lys Gly Lys Gly Ile Ala Asn Ala Ser Ser Val 
        355                 360                 365             


Glu His Tyr Leu Ser Asp Val Cys Tyr Thr Arg Ala Ala Glu Leu Phe 
    370                 375                 380                 


Lys Asn Ala Ala Ile Ala Ser Gly Leu Arg Ser Lys Ile Lys Ser Asn 
385                 390                 395                 400 


Phe Arg Leu Lys Glu Leu Lys Asn Met Lys Ser Gly Leu Pro Thr Thr 
                405                 410                 415     


Lys Ser Asp Asn Phe Pro Ile Pro Leu Val Lys Gln Lys Gly Gly Gln 
            420                 425                 430         


Tyr Thr Gly Phe Glu Ile Ser Asn His Asn Ser Asp Phe Ile Ile Lys 
        435                 440                 445             


Ile Pro Phe Gly Arg Trp Gln Val Lys Lys Glu Ile Asp Lys Tyr Arg 
    450                 455                 460                 


Pro Trp Glu Lys Phe Asp Phe Glu Gln Val Gln Lys Ser Pro Lys Pro 
465                 470                 475                 480 


Ile Ser Leu Leu Leu Ser Thr Gln Arg Arg Lys Arg Asn Lys Gly Trp 
                485                 490                 495     


Ser Lys Asp Glu Gly Thr Glu Ala Glu Ile Lys Lys Val Met Asn Gly 
            500                 505                 510         


Asp Tyr Gln Thr Ser Tyr Ile Glu Val Lys Arg Gly Ser Lys Ile Cys 
        515                 520                 525             


Glu Lys Ser Ala Trp Met Leu Asn Leu Ser Ile Asp Val Pro Lys Ile 
    530                 535                 540                 


Asp Lys Gly Val Asp Pro Ser Ile Ile Gly Gly Ile Ala Val Gly Val 
545                 550                 555                 560 


Lys Ser Pro Leu Val Cys Ala Ile Asn Asn Ala Phe Ser Arg Tyr Ser 
                565                 570                 575     


Ile Ser Asp Asn Asp Leu Phe His Phe Asn Lys Lys Met Phe Ala Arg 
            580                 585                 590         


Arg Arg Ile Leu Leu Lys Lys Asn Arg His Lys Arg Ala Gly His Gly 
        595                 600                 605             


Ala Lys Asn Lys Leu Lys Pro Ile Thr Ile Leu Thr Glu Lys Ser Glu 
    610                 615                 620                 


Arg Phe Arg Lys Lys Leu Ile Glu Arg Trp Ala Cys Glu Ile Ala Asp 
625                 630                 635                 640 


Phe Phe Ile Lys Asn Lys Val Gly Thr Val Gln Met Glu Asn Leu Glu 
                645                 650                 655     


Ser Met Lys Arg Lys Glu Asp Ser Tyr Phe Asn Ile Arg Leu Arg Gly 
            660                 665                 670         


Phe Trp Pro Tyr Ala Glu Met Gln Asn Lys Ile Glu Phe Lys Leu Lys 
        675                 680                 685             


Gln Tyr Gly Ile Glu Ile Arg Lys Val Ala Pro Asn Asn Thr Ser Lys 
    690                 695                 700                 


Thr Cys Ser Lys Cys Gly His Leu Asn Asn Tyr Phe Asn Phe Glu Tyr 
705                 710                 715                 720 


Arg Lys Lys Asn Lys Phe Pro His Phe Lys Cys Glu Lys Cys Asn Phe 
                725                 730                 735     


Lys Glu Asn Ala Asp Tyr Asn Ala Ala Leu Asn Ile Ser Asn Pro Lys 
            740                 745                 750         


Leu Lys Ser Thr Lys Glu Glu Pro Gly Lys Glu Ile Gln Leu Lys Pro 
        755                 760                 765             


Lys Ala Asn Val Ser Ser Tyr Val His Phe Leu Leu Asn Tyr Arg Asn 
    770                 775                 780                 


Lys Phe Lys Glu Gln Gln Pro Asn Thr Tyr Val Gly Phe Lys Glu Phe 
785                 790                 795                 800 


Ser Arg Lys Cys Ser Glu Lys Trp Arg Ser Ile Ser Lys His Glu Lys 
                805                 810                 815     


Ala Lys Tyr Glu Ala Leu Ala Lys Leu Asp Lys Ala Arg Tyr Gln Glu 
            820                 825                 830         


Glu Met Met Asn Tyr Val 
        835             


<210>  282
<211>  225
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dCas14-FokI Siglec SL SCNA

<400>  282
gggcuucacu gauaaagugg agaaccgcuu caccaaaagc ugucccuuag gggauuagaa       60

cuugagugaa ggugggcugc uugcaucagc cuaaugucga gaagugcuuu cuucggaaag      120

uaacccucga aacaaauuca uuuuuccucu ccaauucugc acaagaaagu ugcagaaccc      180

gaauagacga augaaggaau gcaacgggtg gagagaggaa gctat                      225


<210>  283
<211>  225
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dCas14-FokI Siglec ASR SCNA

<400>  283
gggcuucacu gauaaagugg agaaccgcuu caccaaaagc ugucccuuag gggauuagaa       60

cuugagugaa ggugggcugc uugcaucagc cuaaugucga gaagugcuuu cuucggaaag      120

uaacccucga aacaaauuca uuuuuccucu ccaauucugc acaagaaagu ugcagaaccc      180

gaauagacga augaaggaau gcaacagaaa gaacccatcg ttcat                      225


<210>  284
<211>  1602
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Ancestral dCas9-FokI

<400>  284

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Gly Thr Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr 
225                 230                 235                 240 


Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser 
                245                 250                 255     


Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys Lys 
            260                 265                 270         


Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala 
        275                 280                 285             


Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn 
    290                 295                 300                 


Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu 
305                 310                 315                 320 


Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu 
                325                 330                 335     


Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp Glu 
            340                 345                 350         


Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys 
        355                 360                 365             


Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala 
    370                 375                 380                 


Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp 
385                 390                 395                 400 


Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val 
                405                 410                 415     


Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser Gly 
            420                 425                 430         


Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg 
        435                 440                 445             


Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly Leu 
    450                 455                 460                 


Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys 
465                 470                 475                 480 


Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp 
                485                 490                 495     


Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln 
            500                 505                 510         


Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu 
        515                 520                 525             


Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro Leu 
    530                 535                 540                 


Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala 
545                 550                 555                 560 


Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu 
                565                 570                 575     


Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Ile 
            580                 585                 590         


Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu Glu 
        595                 600                 605             


Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu 
    610                 615                 620                 


Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg 
625                 630                 635                 640 


Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu 
                645                 650                 655     


His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Asp 
            660                 665                 670         


Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr 
        675                 680                 685             


Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg 
    690                 695                 700                 


Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp 
705                 710                 715                 720 


Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp 
                725                 730                 735     


Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr 
            740                 745                 750         


Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr 
        755                 760                 765             


Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu Ala 
    770                 775                 780                 


Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln 
785                 790                 795                 800 


Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu 
                805                 810                 815     


Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His 
            820                 825                 830         


Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu 
        835                 840                 845             


Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu 
    850                 855                 860                 


Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu Phe 
865                 870                 875                 880 


Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp 
                885                 890                 895     


Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser 
            900                 905                 910         


Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg 
        915                 920                 925             


Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu 
    930                 935                 940                 


Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln 
945                 950                 955                 960 


Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln 
                965                 970                 975     


Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro 
            980                 985                 990         


Glu Asn Ile Val Ile Glu Met Ala  Arg Glu Asn Gln Thr  Thr Gln Lys 
        995                 1000                 1005             


Gly Gln  Lys Asn Ser Arg Glu  Arg Met Lys Arg Ile  Glu Glu Gly 
    1010                 1015                 1020             


Ile Lys  Glu Leu Gly Ser Gln  Ile Leu Lys Glu His  Pro Val Glu 
    1025                 1030                 1035             


Asn Thr  Gln Leu Gln Asn Glu  Lys Leu Tyr Leu Tyr  Tyr Leu Gln 
    1040                 1045                 1050             


Asn Gly  Lys Asp Met Tyr Thr  Gly Asp Glu Leu Asp  Ile Asn Arg 
    1055                 1060                 1065             


Leu Tyr  Gln Tyr Asp Val Asp  Ala Ile Ile Pro Gln  Ser Phe Ile 
    1070                 1075                 1080             


Lys Asp  Asn Ser Ile Asp Asn  Lys Val Leu Thr Ser  Ser Lys Glu 
    1085                 1090                 1095             


Asn Arg  Gly Lys Ser Asp Asp  Val Pro Ser Glu Glu  Val Val Lys 
    1100                 1105                 1110             


Lys Met  Lys Asn Tyr Trp Arg  Gln Leu Leu Asn Ala  Lys Leu Ile 
    1115                 1120                 1125             


Thr Gln  Arg Lys Phe Asp Asn  Leu Thr Lys Ala Glu  Arg Gly Gly 
    1130                 1135                 1140             


Leu Ser  Glu Ala Asp Lys Ala  Gly Phe Ile Lys Arg  Gln Leu Val 
    1145                 1150                 1155             


Glu Thr  Arg Gln Ile Thr Lys  His Val Ala Gln Ile  Leu Asp Ser 
    1160                 1165                 1170             


Arg Met  Asn Thr Lys Arg Asp  Lys Asn Asp Lys Pro  Ile Arg Glu 
    1175                 1180                 1185             


Val Lys  Val Ile Thr Leu Lys  Ser Lys Leu Val Ser  Asp Phe Arg 
    1190                 1195                 1200             


Lys Asp  Phe Gln Leu Tyr Lys  Val Arg Glu Ile Asn  Asn Tyr His 
    1205                 1210                 1215             


His Ala  His Asp Ala Tyr Leu  Asn Ala Val Val Gly  Thr Ala Leu 
    1220                 1225                 1230             


Ile Lys  Lys Tyr Pro Lys Leu  Glu Ser Glu Phe Val  Tyr Gly Asp 
    1235                 1240                 1245             


Tyr Lys  Val Tyr Asp Val Arg  Lys Met Ile Ala Lys  Ser Glu Gln 
    1250                 1255                 1260             


Glu Ile  Gly Lys Ala Thr Ala  Lys Arg Phe Phe Tyr  Ser Asn Ile 
    1265                 1270                 1275             


Met Asn  Phe Phe Lys Thr Glu  Val Lys Leu Ala Asn  Gly Glu Ile 
    1280                 1285                 1290             


Arg Lys  Arg Pro Leu Ile Glu  Thr Asn Glu Glu Thr  Gly Glu Ile 
    1295                 1300                 1305             


Val Trp  Asp Lys Glu Lys Asp  Phe Ala Thr Val Arg  Lys Val Leu 
    1310                 1315                 1320             


Ser Met  Pro Gln Val Asn Ile  Val Lys Lys Thr Glu  Val Gln Thr 
    1325                 1330                 1335             


Gly Gly  Phe Ser Lys Glu Ser  Ile Leu Ser Lys Gly  Glu Ser Asp 
    1340                 1345                 1350             


Lys Leu  Ile Pro Arg Lys Lys  Gly Trp Asp Ser Lys  Lys Tyr Gly 
    1355                 1360                 1365             


Gly Phe  Gly Ser Pro Thr Val  Ala Tyr Ser Val Leu  Val Val Ala 
    1370                 1375                 1380             


Lys Val  Glu Lys Gly Lys Ala  Lys Lys Leu Lys Thr  Val Lys Glu 
    1385                 1390                 1395             


Leu Val  Gly Ile Thr Ile Met  Glu Lys Ser Ser Phe  Glu Lys Asp 
    1400                 1405                 1410             


Pro Ile  Gly Phe Leu Glu Ala  Lys Gly Tyr Lys Asp  Ile Gln Lys 
    1415                 1420                 1425             


Asp Leu  Ile Ile Lys Leu Pro  Lys Tyr Ser Leu Phe  Glu Leu Glu 
    1430                 1435                 1440             


Asn Gly  Arg Arg Arg Met Leu  Ala Ser Ala Thr Glu  Leu Gln Lys 
    1445                 1450                 1455             


Ala Asn  Glu Leu Val Leu Pro  Gln His Leu Val Thr  Leu Leu Tyr 
    1460                 1465                 1470             


His Ala  His Lys Ile Glu Ser  Ser Lys Glu Ser Glu  Asn Gln Ala 
    1475                 1480                 1485             


Tyr Ile  Glu Gln His Arg His  Glu Phe Asp Glu Ile  Leu Glu Tyr 
    1490                 1495                 1500             


Ile Ile  Asp Phe Ser Glu Lys  Tyr Ile Leu Ala Glu  Lys Asn Ile 
    1505                 1510                 1515             


Ser Lys  Val Lys Ser Leu Phe  Asn Lys Gln Phe Glu  Asp Ser Asp 
    1520                 1525                 1530             


Ile Ser  Glu Leu Ser Asn Ser  Phe Ile Asn Leu Leu  Thr Leu Thr 
    1535                 1540                 1545             


Ser Phe  Gly Ala Pro Ala Ala  Phe Lys Phe Leu Asp  Thr Thr Ile 
    1550                 1555                 1560             


Lys Gln  Gly Arg Lys Arg Tyr  Gln Ser Thr Lys Glu  Val Leu Asp 
    1565                 1570                 1575             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1580                 1585                 1590             


Thr Asp  Leu Ser Gln Leu Gly  Gly Asp 
    1595                 1600         


<210>  285
<211>  154
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  Human CTLA4 exon and 5' untranslated sequence

<400>  285
ttttgctcta cttcctgaag acctgaacac cgctcccata aagccatggc ttgccttgga       60

tttcagcggc acaaggctca gctgaacctg gctaccagga cctggccctg cactctcctg      120

ttttttcttc tcttcatccc tgtcttctgc aaag                                  154


<210>  286
<211>  122
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI CTLA4 SL SCNA

<400>  286
ggcgcgttta ttccattact ttggagccag tcccagcgac tatgtcgtat ggacgaagcg       60

cttatttatc ggagagaaac cgataagtaa aacgcatcaa agugaagacc ugaacaccgc      120

uc                                                                     122


<210>  287
<211>  122
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dCasX-FokI CTLA4 ASR SCNA

<400>  287
ggcgcgttta ttccattact ttggagccag tcccagcgac tatgtcgtat ggacgaagcg       60

cttatttatc ggagagaaac cgataagtaa aacgcatcaa aggccgcuga aauccaaggc      120

aa                                                                     122


<210>  288
<211>  1610
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-CspCI

<400>  288

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Gly Met Leu Asn Pro Pro Tyr Ser Gln Ser Lys Ser Asp Ala Glu 
            20                  25                  30          


Leu His Glu Leu Tyr Phe Val Lys Gln Met Leu Asp Thr Leu Thr Pro 
        35                  40                  45              


Gly Gly Val Gly Ile Ala Ile Val Pro Met Ser Ser Ala Ile Ser Pro 
    50                  55                  60                  


Asn Pro Met Arg Glu Glu Leu Met Lys Tyr His Ser Leu Asp Ala Val 
65                  70                  75                  80  


Met Ser Met Pro Gln Glu Leu Phe Tyr Pro Val Gly Thr Val Thr Cys 
                85                  90                  95      


Val Met Val Trp Ile Ala Gly Val Pro His Glu Gln Met Ser Lys Lys 
            100                 105                 110         


Thr Trp Phe Gly Tyr Trp Arg Asp Asp Gly Phe Val Lys Thr Lys His 
        115                 120                 125             


Lys Gly Arg Ile Asp Met Asn Gly Thr Trp Pro Asp Ile Arg Asp Arg 
    130                 135                 140                 


Trp Ile Glu Met Tyr Arg Asn Arg Glu Val His Ala Gly Glu Ser Ile 
145                 150                 155                 160 


Met Gln Lys Val Gly Pro Asp Asp Glu Trp Cys Ala Glu Ala Tyr Met 
                165                 170                 175     


Glu Thr Asp Tyr Ser Val Leu Thr Gln Ser Asp Phe Glu Lys Val Val 
            180                 185                 190         


Gln Ser Tyr Ala Leu Phe Lys Leu Phe Gly Gln Gly Ser Ser Gln Ser 
        195                 200                 205             


Glu Val Lys Gly Ala Thr Asp Ala Glu Asp Ser Gly Ser Glu Thr Pro 
    210                 215                 220                 


Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr 
225                 230                 235                 240 


Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile 
                245                 250                 255     


Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn 
            260                 265                 270         


Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe 
        275                 280                 285             


Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg 
    290                 295                 300                 


Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile 
305                 310                 315                 320 


Phe Ala Asn Glu Met Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu 
                325                 330                 335     


Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro 
            340                 345                 350         


Ile Phe Gly Asn Leu Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro 
        355                 360                 365             


Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala 
    370                 375                 380                 


Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg 
385                 390                 395                 400 


Gly His Phe Leu Ile Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val 
                405                 410                 415     


Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu 
            420                 425                 430         


Glu Ser Pro Leu Asp Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser 
        435                 440                 445             


Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe 
    450                 455                 460                 


Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala 
465                 470                 475                 480 


Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp 
                485                 490                 495     


Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu 
            500                 505                 510         


Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala 
        515                 520                 525             


Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn 
    530                 535                 540                 


Ser Glu Val Thr Lys Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr 
545                 550                 555                 560 


Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln 
                565                 570                 575     


Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn 
            580                 585                 590         


Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr 
        595                 600                 605             


Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile 
    610                 615                 620                 


Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg 
625                 630                 635                 640 


Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro 
                645                 650                 655     


His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu 
            660                 665                 670         


Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile 
        675                 680                 685             


Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn 
    690                 695                 700                 


Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro 
705                 710                 715                 720 


Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe 
                725                 730                 735     


Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val 
            740                 745                 750         


Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu 
        755                 760                 765             


Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe 
    770                 775                 780                 


Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr 
785                 790                 795                 800 


Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys 
                805                 810                 815     


Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe 
            820                 825                 830         


Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp 
        835                 840                 845             


Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile 
    850                 855                 860                 


Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg 
865                 870                 875                 880 


Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu 
                885                 890                 895     


Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile 
            900                 905                 910         


Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu 
        915                 920                 925             


Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp 
    930                 935                 940                 


Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly 
945                 950                 955                 960 


Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro 
                965                 970                 975     


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
            980                 985                 990         


Val Lys Val Met Gly His Lys Pro  Glu Asn Ile Val Ile  Glu Met Ala 
        995                 1000                 1005             


Arg Glu  Asn Gln Thr Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu 
    1010                 1015                 1020             


Arg Lys  Lys Arg Ile Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln 
    1025                 1030                 1035             


Ile Leu  Lys Glu Asn Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
    1040                 1045                 1050             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1055                 1060                 1065             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1070                 1075                 1080             


Ala Ile  Val Pro Gln Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn 
    1085                 1090                 1095             


Lys Val  Leu Thr Arg Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn 
    1100                 1105                 1110             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1115                 1120                 1125             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1130                 1135                 1140             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala 
    1145                 1150                 1155             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1160                 1165                 1170             


His Val  Ala Arg Ile Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp 
    1175                 1180                 1185             


Lys Asn  Asp Lys Pro Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1190                 1195                 1200             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys 
    1205                 1210                 1215             


Val Arg  Asp Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1220                 1225                 1230             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1235                 1240                 1245             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1250                 1255                 1260             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1265                 1270                 1275             


Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1280                 1285                 1290             


Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1295                 1300                 1305             


Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp 
    1310                 1315                 1320             


Phe Ala  Thr Val Arg Lys Val  Leu Ala Met Pro Gln  Val Asn Ile 
    1325                 1330                 1335             


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1340                 1345                 1350             


Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys 
    1355                 1360                 1365             


Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val 
    1370                 1375                 1380             


Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ala 
    1385                 1390                 1395             


Lys Lys  Leu Lys Ser Val Lys  Val Leu Val Gly Ile  Thr Ile Met 
    1400                 1405                 1410             


Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala 
    1415                 1420                 1425             


Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro 
    1430                 1435                 1440             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu 
    1445                 1450                 1455             


Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro 
    1460                 1465                 1470             


Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala 
    1475                 1480                 1485             


Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu 
    1490                 1495                 1500             


Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys 
    1505                 1510                 1515             


Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe 
    1520                 1525                 1530             


Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser 
    1535                 1540                 1545             


Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly 
    1550                 1555                 1560             


Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr 
    1565                 1570                 1575             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1580                 1585                 1590             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1595                 1600                 1605             


Gly Asp  
    1610 


<210>  289
<211>  1586
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-BsgI

<400>  289

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Phe Lys Ser Glu Phe Glu Leu Asn Met Ala Arg His Leu Ser Glu 
            20                  25                  30          


Met Glu Ile Leu Asn Tyr Ile Thr Glu Phe Glu Ile Gly Thr Gly Asn 
        35                  40                  45              


Phe Val Trp Asn Gln Asn Lys Asp Asp Ile Tyr Asn Gln Tyr Val Glu 
    50                  55                  60                  


Asp Ser Ile Pro Leu Ile Met Gly Pro Ser Ile Asn Leu Asp Gly Ile 
65                  70                  75                  80  


Asn Leu Glu Asn Ser Lys Tyr Asn Tyr Cys Val Val Ser Asn Glu Asn 
                85                  90                  95      


Lys Tyr Ile Arg Asn Asp Ile Val Ile Leu Tyr Arg Arg Met Ser Pro 
            100                 105                 110         


Ile Gly Asn Glu Ile Arg Met Ile Ala Thr Ile Ile Asn Thr Asp Ser 
        115                 120                 125             


Ile Pro Lys Tyr Val Leu Glu Asn His Val Asn Phe Ile Lys His Glu 
    130                 135                 140                 


Asn Tyr Asn Met Arg Lys Ile Gln Met Met Leu Gln Phe Ile Thr Ser 
145                 150                 155                 160 


Glu Asp Phe Glu Thr Met Ile Asp Ile Phe Cys Gln Thr Asn Gln Val 
                165                 170                 175     


Ser Ser Asn Asp Leu Phe Thr Ile Phe Glu Val Leu Met Ser Leu Arg 
            180                 185                 190         


Asn His Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
        195                 200                 205             


Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr 
    210                 215                 220                 


Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser 
225                 230                 235                 240 


Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys 
                245                 250                 255     


Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala 
            260                 265                 270         


Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn 
        275                 280                 285             


Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu 
    290                 295                 300                 


Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu 
305                 310                 315                 320 


Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu 
                325                 330                 335     


Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys 
            340                 345                 350         


Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala 
        355                 360                 365             


Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys 
    370                 375                 380                 


Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile 
385                 390                 395                 400 


Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu 
                405                 410                 415     


Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg 
            420                 425                 430         


Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu 
        435                 440                 445             


Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys 
    450                 455                 460                 


Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp 
465                 470                 475                 480 


Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln 
                485                 490                 495     


Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu 
            500                 505                 510         


Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu 
        515                 520                 525             


Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala 
    530                 535                 540                 


Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu 
545                 550                 555                 560 


Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile 
                565                 570                 575     


Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu 
            580                 585                 590         


Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu 
        595                 600                 605             


Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg 
    610                 615                 620                 


Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu 
625                 630                 635                 640 


His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu 
                645                 650                 655     


Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr 
            660                 665                 670         


Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg 
        675                 680                 685             


Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp 
    690                 695                 700                 


Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp 
705                 710                 715                 720 


Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr 
                725                 730                 735     


Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr 
            740                 745                 750         


Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala 
        755                 760                 765             


Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln 
    770                 775                 780                 


Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu 
785                 790                 795                 800 


Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His 
                805                 810                 815     


Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu 
            820                 825                 830         


Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu 
        835                 840                 845             


Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe 
    850                 855                 860                 


Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp 
865                 870                 875                 880 


Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser 
                885                 890                 895     


Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg 
            900                 905                 910         


Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu 
        915                 920                 925             


Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln 
    930                 935                 940                 


Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln 
945                 950                 955                 960 


Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro 
                965                 970                 975     


Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys 
            980                 985                 990         


Gly Leu Gln Gln Ser Arg Glu Arg  Lys Lys Arg Ile Glu  Glu Gly Ile 
        995                 1000                 1005             


Lys Glu  Leu Glu Ser Gln Ile  Leu Lys Glu Asn Pro  Val Glu Asn 
    1010                 1015                 1020             


Thr Gln  Leu Gln Asn Glu Lys  Leu Tyr Leu Tyr Tyr  Leu Gln Asn 
    1025                 1030                 1035             


Gly Arg  Asp Met Tyr Val Asp  Gln Glu Leu Asp Ile  Asn Arg Leu 
    1040                 1045                 1050             


Ser Asp  Tyr Asp Val Asp Ala  Ile Val Pro Gln Ser  Phe Ile Lys 
    1055                 1060                 1065             


Asp Asp  Ser Ile Asp Asn Lys  Val Leu Thr Arg Ser  Val Glu Asn 
    1070                 1075                 1080             


Arg Gly  Lys Ser Asp Asn Val  Pro Ser Glu Glu Val  Val Lys Lys 
    1085                 1090                 1095             


Met Lys  Asn Tyr Trp Arg Gln  Leu Leu Asn Ala Lys  Leu Ile Thr 
    1100                 1105                 1110             


Gln Arg  Lys Phe Asp Asn Leu  Thr Lys Ala Glu Arg  Gly Gly Leu 
    1115                 1120                 1125             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1130                 1135                 1140             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1145                 1150                 1155             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1160                 1165                 1170             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1175                 1180                 1185             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1190                 1195                 1200             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1205                 1210                 1215             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1220                 1225                 1230             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1235                 1240                 1245             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1250                 1255                 1260             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1265                 1270                 1275             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1280                 1285                 1290             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1295                 1300                 1305             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1310                 1315                 1320             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1325                 1330                 1335             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1340                 1345                 1350             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1355                 1360                 1365             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1370                 1375                 1380             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1385                 1390                 1395             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1400                 1405                 1410             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1415                 1420                 1425             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1430                 1435                 1440             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1445                 1450                 1455             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1460                 1465                 1470             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1475                 1480                 1485             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1490                 1495                 1500             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1505                 1510                 1515             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1520                 1525                 1530             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1535                 1540                 1545             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1550                 1555                 1560             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1565                 1570                 1575             


Asp Leu  Ser Gln Leu Gly Gly  Asp 
    1580                 1585     


<210>  290
<211>  1565
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-BbvI

<400>  290

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Ala Leu Asn His Glu Phe Thr Arg His Lys Lys Lys Val Val Glu Met 
            20                  25                  30          


Leu Asn His Ile Ala Pro Gly Gly Asn Ala Trp Ser Glu Ser Ile Pro 
        35                  40                  45              


Glu Glu Leu Arg Leu Asn Val Lys Lys Val Arg Met Ser Gln Ile Tyr 
    50                  55                  60                  


Arg Arg Leu His Pro Asp Gln Pro Ser Tyr Thr Val Thr Gly Ser Gly 
65                  70                  75                  80  


Gly Gly Gly Thr His Gly Tyr His Trp Glu Glu Pro Arg Ala Leu Thr 
                85                  90                  95      


Asn Arg Glu Arg Ala Arg Leu Gln Thr Phe Pro Asp Asp Tyr Glu Phe 
            100                 105                 110         


Ile Gly Lys Lys Glu Met Val Arg Lys Gln Ile Gly Met Ala Val Pro 
        115                 120                 125             


Pro Asp Gly Ala Lys Ile Ile Leu Glu Ala Val Leu Lys Thr Phe Ala 
    130                 135                 140                 


Arg Ile Glu Tyr Pro Ser Ile Asn Ser Lys Trp Asp Phe Glu Ser Val 
145                 150                 155                 160 


Ser Ala Glu Gln Val Ile Glu Glu Val Gln Glu Ile Met Ser Gly Ser 
                165                 170                 175     


Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Thr Glu 
            180                 185                 190         


Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp 
        195                 200                 205             


Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Phe Lys Val 
    210                 215                 220                 


Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu Met Gly Ala 
225                 230                 235                 240 


Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg 
                245                 250                 255     


Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Arg Tyr Leu 
            260                 265                 270         


Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp Ser Phe Phe 
        275                 280                 285             


Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys Asn Glu 
    290                 295                 300                 


Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala Tyr His Arg 
305                 310                 315                 320 


Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp Ser Pro 
                325                 330                 335     


Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Ile Ile 
            340                 345                 350         


Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn Ala Glu Asn 
        355                 360                 365             


Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr Tyr Asn Gln 
    370                 375                 380                 


Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp Ala Lys Gly 
385                 390                 395                 400 


Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu Lys Leu Ile 
                405                 410                 415     


Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly Asn Ile Ile 
            420                 425                 430         


Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu 
        435                 440                 445             


Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp 
    450                 455                 460                 


Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe 
465                 470                 475                 480 


Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu 
                485                 490                 495     


Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala Ser Met Val 
            500                 505                 510         


Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu Lys Thr Leu 
        515                 520                 525             


Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe Lys Asp Asp 
    530                 535                 540                 


Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile Lys His Arg 
545                 550                 555                 560 


Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr Lys Phe Ile 
                565                 570                 575     


Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys 
            580                 585                 590         


Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly 
        595                 600                 605             


Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu Arg 
    610                 615                 620                 


Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys Ile 
625                 630                 635                 640 


Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala 
                645                 650                 655     


Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu Ala 
            660                 665                 670         


Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala 
        675                 680                 685             


Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn 
    690                 695                 700                 


Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val 
705                 710                 715                 720 


Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys 
                725                 730                 735     


Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu 
            740                 745                 750         


Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr 
        755                 760                 765             


Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu 
    770                 775                 780                 


Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile 
785                 790                 795                 800 


Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu 
                805                 810                 815     


Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile 
            820                 825                 830         


Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met 
        835                 840                 845             


Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg 
    850                 855                 860                 


Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu 
865                 870                 875                 880 


Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu 
                885                 890                 895     


Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln 
            900                 905                 910         


Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala 
        915                 920                 925             


Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val 
    930                 935                 940                 


Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile 
945                 950                 955                 960 


Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly Leu Gln Gln Ser 
                965                 970                 975     


Arg Glu Arg Lys Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Glu Ser 
            980                 985                 990         


Gln Ile Leu Lys Glu Asn Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
        995                 1000                 1005             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1010                 1015                 1020             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1025                 1030                 1035             


Ala Ile  Val Pro Gln Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn 
    1040                 1045                 1050             


Lys Val  Leu Thr Arg Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn 
    1055                 1060                 1065             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1070                 1075                 1080             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1085                 1090                 1095             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala 
    1100                 1105                 1110             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1115                 1120                 1125             


His Val  Ala Arg Ile Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp 
    1130                 1135                 1140             


Lys Asn  Asp Lys Pro Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1145                 1150                 1155             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys 
    1160                 1165                 1170             


Val Arg  Asp Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1175                 1180                 1185             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1190                 1195                 1200             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1205                 1210                 1215             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1220                 1225                 1230             


Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1235                 1240                 1245             


Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1250                 1255                 1260             


Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp 
    1265                 1270                 1275             


Phe Ala  Thr Val Arg Lys Val  Leu Ala Met Pro Gln  Val Asn Ile 
    1280                 1285                 1290             


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1295                 1300                 1305             


Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys 
    1310                 1315                 1320             


Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val 
    1325                 1330                 1335             


Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ala 
    1340                 1345                 1350             


Lys Lys  Leu Lys Ser Val Lys  Val Leu Val Gly Ile  Thr Ile Met 
    1355                 1360                 1365             


Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala 
    1370                 1375                 1380             


Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro 
    1385                 1390                 1395             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu 
    1400                 1405                 1410             


Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro 
    1415                 1420                 1425             


Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala 
    1430                 1435                 1440             


Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu 
    1445                 1450                 1455             


Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys 
    1460                 1465                 1470             


Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe 
    1475                 1480                 1485             


Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser 
    1490                 1495                 1500             


Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly 
    1505                 1510                 1515             


Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr 
    1520                 1525                 1530             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1535                 1540                 1545             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1550                 1555                 1560             


Gly Asp  
    1565 


<210>  291
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  ScCas9 "Sc loop"

<400>  291

Ile Lys His Arg Lys Arg Thr Thr Lys Leu 
1               5                   10  


<210>  292
<211>  58
<212>  PRT
<213>  Streptococcus canis

<400>  292

Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln 
1               5                   10                  15      


Glu Ile Gly Lys Ala Thr Ala Lys Arg Phe Phe Tyr Ser Asn Ile Met 
            20                  25                  30          


Asn Phe Phe Lys Thr Glu Val Lys Leu Ala Asn Gly Glu Ile Arg Lys 
        35                  40                  45              


Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr 
    50                  55              


<210>  293
<211>  3814
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CAR-T NY-ESO-1 cassette for integration at PD-1 locus

<400>  293
agagggggcc acccaccttc cccaggcagg gaggcccggc ccccagggag atggggggga       60

tgggggagga gaagggcctg cccccacccg gcagcctcag gaggggcagc tcgggcggga      120

tatggaaaga ggccacagca gtgagcagag acacagagga ggaaggggcc ctgagctggg      180

gagaccccca cggggtaggg cgtgggggcc acgggcccac ctcctcccca tctcctctgt      240

ctccctgtct ctgtctctct ctccctcccc caccctctcc ccagtcctac cccctcctca      300

cccctcctcc cccagcactg cctctgtcac tctcgcccac gtggatgtgg aggaagaggg      360

ggcgggagca aggggcgggc accctccctt caacctgacc tgggacagtt tcccttccgc      420

tcacctccgc ctgagcagtg gagaaggcgg cactctggtg gggctgctcc aggcatgcag      480

atcccacagg cgccctggcc ttcatagccc atatatggag ttccgcgtta cataacttac      540

ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac      600

gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt      660

acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat      720

tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga      780

ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt      840

ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca      900

ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg      960

tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta     1020

tataagcaga gctggtttag tgaaccgtca gatcctcgag atgctgctgc tggtgaccag     1080

cctgctgctg tgtgagctgc cccaccccgc ctttctggaa gtacaacttc ttgagagcgg     1140

agggggcctc gtgcaacctg gggggtcact gcgactttcc tgcgccgcat ccggcttcac     1200

gtttagcacg taccaaatgt cctgggtccg acaggcccca ggcaaaggcc tcgaatgggt     1260

gagtggcatt gtgagcagcg gcggtagtac cgcatatgct gactccgtca agggccggtt     1320

caccatctcc cgcgataatt ccaagaatac cctgtatttg caaatgaata gtttgagagc     1380

tgaagacact gccgtctatt attgcgcggg cgaactcctg ccttactatg gtatggacgt     1440

gtggggtcag ggtaccaccg taacagtctc ttcaggtggt ggaggatctg ggggcggagg     1500

atccggtgga ggaggatcac aatccgaact tacgcagcct cgatccgtat ccgggagccc     1560

tgggcagagt gtgacaatat cctgtaccgg gacatcaagg gatgtaggcg gatataacta     1620

cgtctcttgg taccaacaac acccgggtaa ggctccgaag ctgataatcc atgacgtgat     1680

agagcgatcc tccggggtgc cggatagatt ctcagggtcc aaaagtggca atacagctag     1740

tctcacaata tcaggactcc aagctgaaga tgaggccgat tattactgtt ggtcctttgc     1800

agggtcttat tatgtttttg ggacaggaac tgatgtcacg gtgctcggta gcgagagcaa     1860

gtacggccct ccctgccccc cttgccctgc ccccgagttc ctgggcggac ccagcgtgtt     1920

cctgttcccc cccaagccca aggacaccct gatgatcagc cggacccccg aggtgacctg     1980

tgtggtggtg gacgtgtccc aggaggaccc cgaggtccag ttcaactggt acgtggacgg     2040

cgtggaggtg cacaacgcca agaccaagcc ccgggaggag cagttcaata gcacctaccg     2100

ggtggtgtcc gtgctgaccg tgctgcacca ggactggctg aacggcaagg aatacaagtg     2160

taaggtgtcc aacaagggcc tgcccagcag catcgagaaa accatcagca aggccaaggg     2220

ccagcctcgg gagccccagg tgtacaccct gccccctagc caagaggaga tgaccaagaa     2280

tcaggtgtcc ctgacctgcc tggtgaaggg cttctacccc agcgacatcg ccgtggagtg     2340

ggagagcaac ggccagcccg agaacaacta caagaccacc ccccctgtgc tggacagcga     2400

cggcagcttc ttcctgtaca gcaggctgac cgtggacaag agccggtggc aggagggcaa     2460

cgtctttagc tgctccgtga tgcacgaggc cctgcacaac cactacaccc agaagagcct     2520

gtccctgagc ctgggcaaga tgatctacat ctgggcccct ctggccggca cctgtggcgt     2580

gctgctgctg agcctggtca tcaccctgta ctgcaaccac cggaatagga gcaagcggag     2640

cagaggcggc cacagcgact acatgaacat gaccccccgg aggcctggcc ccacccggaa     2700

gcactaccag ccctacgccc ctcccaggga cttcgccgcc taccggagcc gggtgaagtt     2760

cagccggagc gccgacgccc ctgcctacca gcagggccag aaccagctgt acaacgagct     2820

gaacctgggc cggagggagg agtacgacgt gctggacaag cggagaggcc gggaccctga     2880

gatgggcggc aagccccgga gaaagaaccc tcaggagggc ctgtataacg aactgcagaa     2940

agacaagatg gccgaggcct acagcgagat cggcatgaag ggcgagcggc ggaggggcaa     3000

gggccacgac ggcctgtacc agggcctgag caccgccacc aaggatacct acgacgccct     3060

gcacatgcag gccctgcccc ccagatgagt gtgccttcta gttgccagcc atctgttgtt     3120

tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa     3180

taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg     3240

gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg     3300

gtgggctcta tggcgctaca actgggctgg cggccaggat ggttcttagg taggtggggt     3360

cggcggtcag gtgtcccaga gccaggggtc tggagggacc ttccaccctc agtccctggc     3420

aggtcggggg gtgctgaggc gggcctggcc ctggcagccc aggggtcccg gagcgagggg     3480

tctggaggga cctttcactc tcagtccctg gcaggtcggg gggtgctgtg gcaggcccag     3540

ccttggcccc cagctctgcc ccttaccctg agctgtgtgg ctttgggcag ctcgaactcc     3600

tgggttcctc tctgggcccc aactcctccc ctggcccaag tcccctcttt gctcctgggc     3660

aggcaggacc tctgtcccct ctcagccggt ccttggggct gcgtgtttct gtagaatgac     3720

gggtcaggct ggccagaacc ccaaaccttg gccgtgggga gtctgcgtgg cggctctgcc     3780

ttgcccaggc atccttggtc ctcactcgag tttt                                 3814


<210>  294
<211>  6370
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI -EMX1sgRNA15T1-NNG

<400>  294
atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata       60

acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat      120

aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga      180

gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc      240

ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt      300

atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat      360

gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag      420

tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc      480

aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga      540

ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtatgg      600

cccctaagaa gaagagaaaa gtcggcggct ctggcagcca gctggtcaag tctgagctgg      660

aagagaagaa aagcgagctg cggcacaagc tgaaatacgt gccccacgag tacatcgagc      720

tgatcgagat cgccagaaac agcacccagg acagaatcct ggaaatgaag gtcatggaat      780

tctttatgaa ggtgtacggc taccggggca agcacctcgg aggatctaga aagcctgacg      840

gcgccatcta taccgtgggc agccctatcg attacggcgt gatcgtggat accaaggcct      900

acagcggcgg ctacaatctg cctattggac aggccgacga gatgcagaga tacgtggaag      960

aaaaccagac acggaacaag cacatcaacc ccaacgagtg gtggaaagtg taccccagca     1020

gcgtgaccga gttcaagttc ctgttcgtgt ccggccactt caagggcaac tacaaggccc     1080

agctgacccg gctgaaccac atcacaaatt gcaacggcgc tgtgctgtct gtggaagaac     1140

tgctgatcgg cggcgagatg atcaaggccg gaacactgac actggaagaa gtgcggcgga     1200

agttcaacaa cggcgagatc aacttcagcg gcagcgagac acctggcaca tctgaatctg     1260

ccacacctga gagcatggaa aagaagtaca gcatcggcct ggccatcggc acaaattctg     1320

ttggctgggc cgtgatcacc gacgactaca aggtgcccag caagaaattc aaggtgctgg     1380

gcaacaccaa ccggaagtcc atcaagaaaa acctgatggg cgccctgctg ttcgattctg     1440

gcgaaacagc cgaggccacc agactgaaga gaaccgccag aagaagatac acccggcgga     1500

agaaccggat cagatacctg caagagatct tcgccaacga gatggccaag ctggacgaca     1560

gcttctttca gagactggaa gagtccttcc tggtggaaga ggacaagaag aacgagcggc     1620

accccatctt cggcaatctg gctgatgagg tggcctacca cagaaactac cccaccatct     1680

accacctgag gaagaagctg gccgactctc ctgagaaggc cgacctgaga ctgatctatc     1740

tggccctggc tcacatcatc aagttccggg gccacttcct gatcgagggc aagctgaatg     1800

ccgagaacag cgacgtggcc aaactgttct accagctgat ccagacctac aaccagctgt     1860

tcgaggaaag ccctctggac gagattgagg tggacgccaa gggaatcctg agcgccagac     1920

tgagcaagag caagcggctg gaaaagctga tcgccgtgtt tcccaacgag aagaagaatg     1980

gcctgttcgg caacatcatt gccctggcac tgggcctgac acctaacttc aagtccaact     2040

tcgacctgac cgaggacgcc aaactgcagc tgagcaagga cacctacgac gacgatctgg     2100

atgagctgct gggccagatc ggagatcagt acgccgatct gttcagcgcc gccaagaatc     2160

tgtctgacgc catcctgctg agcgacatcc tgagatccaa cagcgaagtg acaaaggccc     2220

ctctgagcgc ctccatggtc aagagatatg acgagcacca ccaggacctg gctctgctga     2280

aaacccttgt gcggcagcag ttccccgaga agtatgccga gatcttcaag gacgacacca     2340

agaacggcta cgccggctat gttggcatcg gcatcaagca cagaaagcgg accaccaagc     2400

tggccacaca agaggaattc tacaagttca tcaagcccat cctcgagaag atggacggcg     2460

ccgaagaact cctggctaag ctgaacaggg acgacctgct gcggaagcag agaaccttcg     2520

acaatggcag catccctcac cagatccacc tgaaagagct gcacgccatt ctgcggcggc     2580

aagaagagtt ctacccattc ctgaaagaaa accgggaaaa gattgagaag atcctgacct     2640

tcaggatccc ctactacgtg ggaccactgg ccagaggcaa tagcagattc gcttggctga     2700

ccagaaagag cgaggaagcc atcacaccct ggaacttcga ggaagtggtg gataagggcg     2760

ccagcgctca gtccttcatc gagcggatga ccaatttcga cgagcagctg cccaacaaga     2820

aggtgctgcc taagcacagc ctgctgtacg agtacttcac cgtgtacaat gagctgacca     2880

aagtcaaata cgtgacagag cggatgcgga agcctgagtt tctgagcggc gagcagaaaa     2940

aggccatcgt ggatctgctg ttcaagacca acagaaaagt gaccgtgaag cagctgaaag     3000

aggactactt caagaaaatc gagtgcttcg acagcgtcga gatcatcggc gtggaagata     3060

gattcaacgc cagcctgggc acataccacg atctgctgaa gattatcaag gacaaggact     3120

tcctggacaa cgaggaaaac gaggacatcc tggaagatat cgtgctgacc ctgacactgt     3180

ttgaggacag agagatgatc gaggaacggc tgaaaacata cgcccacctg ttcgacgaca     3240

aagtgatgaa gcaactgaag cggcggcact acacaggctg gggcagactg tccagaaaga     3300

tgattaacgg catccgggac aagcagagcg gcaagaccat cctggacttt ctgaagtccg     3360

acggcttcag caaccggaac ttcatgcagc tgattcacga cgacagcctc accttcaaag     3420

aggaaatcga gaaggcccag gtttccggcc agggcgattc tctgcacgag cagattgctg     3480

atctggccgg atctcccgcc attaagaagg gcatcctgca gacagtgaag atcgtggacg     3540

aactcgtgaa agtgatgggc cacaagcctg agaacatcgt gatcgaaatg gccagagaga     3600

accagaccac caccaagggc ctgcagcagt ccagagagag aaagaagcgg atcgaagagg     3660

gcatcaaaga gctggaaagc cagatcctca aagagaaccc cgtcgagaac acccagctgc     3720

agaacgagaa gctgtacctg tactacctgc agaatggacg ggatatgtac gtggaccaag     3780

aactggacat caaccggctg tccgactacg atgtggatgc catcgtgccc cagagcttca     3840

tcaaggatga cagcatcgac aacaaggtcc tgaccagatc cgtggaaaac agaggcaaga     3900

gcgacaacgt gccctccgaa gaggtggtca agaagatgaa gaattactgg cgacagctgc     3960

tgaacgccaa gctgatcacc cagagaaagt tcgacaacct gaccaaggcc gagagaggcg     4020

gactgtctga agctgataag gccggcttca ttaagcggca gctggtggaa acccggcaga     4080

tcacaaagca cgtggcccgg atcctggaca gcagaatgaa caccaagcgg gacaagaacg     4140

acaagcctat ccgggaagtg aaagtcatca ccctgaagtc caagctggtg tccgatttcc     4200

ggaaggattt ccagctgtac aaagtgcgcg acatcaacaa ctaccatcac gcccacgacg     4260

cctacctgaa tgctgttgtg ggaacagccc tgatcaagaa gtaccctaag ctggaatccg     4320

agttcgtgta cggcgactac aaagtgtacg acgtgcggaa gatgatcgcc aagagcgagc     4380

aagagattgg caaggccacc gccaagcggt tcttctactc caacatcatg aactttttca     4440

agaccgaagt gaagctcgcc aacggcgaaa tccggaagag gcccctgatt gagacaaatg     4500

gcgagacagg cgaggtcgtg tggaacaaag agaaggactt cgccaccgtg cggaaagtgc     4560

tggccatgcc tcaagtgaat atcgtgaaaa agaccgaggt gcagaccggc ggcttctcca     4620

aagagagcat cctgtctaag agagagagcg ccaagctcat ccctcggaag aaaggctggg     4680

atacacggaa gtacggcgga tttggaagcc ctaccgtggc ctactctatt ctggtggtgg     4740

ccaaggtgga aaagggcaaa gccaagaaac tgaagtctgt gaaggtgctc gtggggatca     4800

ccatcatgga aaaaggcagc tacgagaagg accctatcgg ctttctggaa gccaagggct     4860

acaaggacat caaaaaagag ctgatcttta agctccccaa gtacagcctg ttcgaactgg     4920

aaaacggcag acgcagaatg ctggcctctg ccacagaact gcagaaagcc aatgaactgg     4980

tgctgccaca acacctcgtg cggctgctgt actacaccca gaatatcagc gccaccaccg     5040

gcagcaacaa cctgggatat atcgagcagc accgggaaga gttcaaagag attttcgaga     5100

aaatcatcga cttcagcgag aagtacatcc tgaagaacaa agtgaacagc aacctcaaga     5160

gcagcttcga tgagcagttc gccgtgtccg actctatcct gctgtctaac agcttcgtgt     5220

ccctgctgaa gtacacctcc tttggcgcct ctggcggctt cacctttctg gatctggacg     5280

tgaaacaggg gagactgaga taccagaccg tgactgaggt gctggacgcc actctgatct     5340

accagagcat caccggcctg tacgagacaa gaaccgatct ctctcagctc ggaggcgact     5400

aaaagcttct gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc     5460

cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc     5520

gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg     5580

ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggacaaacc     5640

cacaactacg cgtaaaaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga     5700

ctagccttat tttaacttgc tatttctagc tctaaaactc ctctagaaac tcgtagaggg     5760

tgtttcgtgg tctcatacag aacttataag attcccaaat ccaaagacat ttcacgttta     5820

tggtgatttc ccagaacaca tagcgacatg caaatattgc agggcgccac tcccctgtcc     5880

ctcacagcca tcttcctgcc agggcgcacg cgcgctgggt gttcccgcct agtgacactg     5940

ggcccgcgat tccttggagc gggttgatga cgtcagcgtt caaagagggc ctatttccca     6000

tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt     6060

tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg     6120

ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt     6180

gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac acctgcactc     6240

ctccaccttg gctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc     6300

aacttgaaaa agtggcaccg agtcggtgct ttttttacgc gtacttgcgg ccgcatattc     6360

ctaggatatt                                                            6370


<210>  295
<211>  8890
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pPZP-RCS

<400>  295
ctacttgcat tacagcttac gaaccgaacg aggcttatgt ccactgggtt cgtgcccgaa       60

ttgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc tccgcgagat      120

catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat cggtaacatg      180

agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg      240

cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt      300

ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg      360

acgtttttaa tgtactgaat taacgccgaa ttgaattcct cgagtacgta ggatccattt      420

aaattctaga ggcgcgccgc ggccgctcct gctgagcctc gacatgttgt cgcaaaattc      480

gccctggacc cgcccaacga tttgtcgtca ctgtcaaggt ttgacctgca cttcatttgg      540

ggcccacata caccaaaaaa atgctgcata attctcgggg cagcaagtcg gttacccggc      600

cgccgtgctg gaccgggttg aatggtgccc gtaactttcg gtagagcgga cggccaatac      660

tcaacttcaa ggaatctcac ccatgcgcgc cggcggggaa ccggagttcc cttcagtgag      720

cgttattagt tcgccgctcg gtgtgtcgta gatactagcc cctggggcac ttttgaaatt      780

tgaataagat ttatgtaatc agtcttttag gtttgaccgg ttctgccgct ttttttaaaa      840

ttggatttgt aataataaaa cgcaattgtt tgttattgtg gcgctctatc atagatgtcg      900

ctataaacct attcagcaca atatattgtt ttcattttaa tattgtacat ataagtagta      960

gggtacaatc agtaaattga acggagaata ttattcataa aaatacgata gtaacgggtg     1020

atatattcat tagaatgaac cgaaaccggc ggtaaggatc tgagctacac atgctcaggt     1080

tttttacaac gtgcacaaca gaattgaaag caaatatcat gcgatcatag gcgtctcgca     1140

tatctcatta aagcaggact ctagggacta gttctagact aggtggatcc tcagaagaac     1200

tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc     1260

acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac     1320

gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag     1380

cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatca     1440

tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga     1500

tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc     1560

tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc     1620

cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg     1680

agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg     1740

tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg     1800

tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc     1860

tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca     1920

tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca     1980

atcatgtcga ctgcagaatt cgaagcttga gctcgagatc tgagtccgga ttggtagatt     2040

gcaaatataa tggtttggcg attatccttg aggccacacc tttaaataga tcaatgaatg     2100

ggcaagtatt gacttgcgat aatcctttaa ctttctgcct accatcaatg tggatgagtt     2160

gtcggtaaaa aggatcttac aggccaaatt cgctcttagc cgtacaatat tactcaccgg     2220

tgcgatgccc cccatcgtag gtgaaggtgg aaattaatga tccatcttga gaccacaggc     2280

ccacaacagc taccagtttc ctcaagggtc caccaaaaac gtaagcgctt acgtacatgg     2340

tcgataagaa aaggcaattt gtagatgtta acatccaacg tcgctttcag cccggtggcg     2400

cgccgatatc ctctcttaag gtagcgagct cttaattaat agggataaca gggtaattaa     2460

ctataacggt cctaaggtag cgatggcaaa cagctattat gggtattatg ggtggttctt     2520

tatgcggaca ctgacggctt tatgcctgca ggtcgcgagc gatcgcggta ccgcccgggc     2580

gtcgacaggc ctaagcttag cttgagcttg gatcagattg tcgtttcccg ccttcagttt     2640

aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat     2700

tagaataacg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg     2760

catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct     2820

atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca     2880

agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt     2940

gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca     3000

agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga     3060

ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca     3120

ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg     3180

acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca     3240

ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg     3300

acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg     3360

agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg     3420

tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga     3480

tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga     3540

ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg     3600

gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac     3660

gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac     3720

cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt     3780

ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg     3840

gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt     3900

tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca     3960

aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc     4020

aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg     4080

ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa     4140

ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc     4200

cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg     4260

atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc     4320

accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa     4380

gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag     4440

gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac     4500

ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc     4560

cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta     4620

aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca     4680

gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc     4740

agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca     4800

ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa     4860

atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc     4920

accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc     4980

tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga     5040

cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga     5100

gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg     5160

tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc     5220

cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc     5280

gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg     5340

tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca     5400

cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact     5460

gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa     5520

gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga     5580

tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt     5640

tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga     5700

agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga     5760

gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct     5820

gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct     5880

ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg     5940

cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc     6000

aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt     6060

catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca     6120

gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga     6180

tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa     6240

cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa     6300

aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc     6360

ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg     6420

gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc     6480

aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc     6540

actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg     6600

aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg     6660

ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca     6720

tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca     6780

gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa     6840

ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg     6900

gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg     6960

ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa     7020

ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg     7080

acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc     7140

tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc     7200

ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc     7260

ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg     7320

ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc     7380

actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga     7440

gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc     7500

tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac     7560

caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg     7620

atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc     7680

acgttaaggg attttggtca tgcatgatat atctcccaat ttgtgtaggg cttattatgc     7740

acgcttaaaa ataataaaag cagacttgac ctgatagttt ggctgtgagc aattatgtgc     7800

ttagtgcatc taatcgcttg agttaacgcc ggcgaagcgg cgtcggcttg aacgaatttc     7860

tagctagaca ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt     7920

cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc     7980

tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag     8040

cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa     8100

gcactacatt tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt     8160

catttagcgc ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg     8220

gacctaccaa ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt     8280

cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt     8340

gcagttcgcg cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga     8400

cttctacagc gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt     8460

tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa     8520

tatcactgtg tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca     8580

acgtcggttc gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt     8640

cggcgatcac cgcttccccc atgatgttta actttgtttt agggcgactg ccctgctgcg     8700

taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct     8760

tggatgcccg aggcatagac tgtaccccaa aaaaacatgt cataacaaga agccatgaaa     8820

accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgac     8880

ggcagttacg                                                            8890


<210>  296
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  COMP exon 13 NGS Primer, forward

<400>  296
agggatggag acggacatca                                                   20


<210>  297
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  COMP exon 13 NGS Primer, reverse

<400>  297
ctcggtgggc taaagtcagg                                                   20


<210>  298
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CTLA4 exon 1 NGS Primer, forward

<400>  298
ttgctctact tcctgaagac ct                                                22


<210>  299
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TLA4 exon 1 NGS Primer, reverse

<400>  299
agtctcactc acctttgcag                                                   20


<210>  300
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SIGLEC10 exon 2 NGS Primer, forward

<400>  300
cagatgcagg atgagtcaca gt                                                22


<210>  301
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SIGLEC10 exon 2 NGS Primer, reverse

<400>  301
ccaccccatt ccatacctgt ta                                                22


<210>  302
<211>  16
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ITGB3 exon 1 NGS Primer, forward

<400>  302
cgagatgcga gcgcgg                                                       16


<210>  303
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ITGB3 exon 1 NGS Primer, reverse

<400>  303
cggctcaccc gagacct                                                      17


<210>  304
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RP exon 1 NGS Primer, forward

<400>  304
cttcgcagca ttcttgggtg                                                   20


<210>  305
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RP exon 1 NGS Primer, reverse

<400>  305
agcgtgagga agttgatggg                                                   20


<210>  306
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  long linker

<400>  306

Gly Gly Ser Ala Gly Gly Thr Gly Gly Ser Gly Gly 
1               5                   10          


<210>  307
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  positively charged linker

<400>  307

Gly Gly Lys Ala Arg Gly Lys Gly Gly Ser Gly Gly 
1               5                   10          


<210>  308
<211>  529
<212>  PRT
<213>  Uncultured archaeon


<220>
<221>  MISC_FEATURE
<223>  Cas14a1 MK005734

<400>  308

Met Ala Lys Asn Thr Ile Thr Lys Thr Leu Lys Leu Arg Ile Val Arg 
1               5                   10                  15      


Pro Tyr Asn Ser Ala Glu Val Glu Lys Ile Val Ala Asp Glu Lys Asn 
            20                  25                  30          


Asn Arg Glu Lys Ile Ala Leu Glu Lys Asn Lys Asp Lys Val Lys Glu 
        35                  40                  45              


Ala Cys Ser Lys His Leu Lys Val Ala Ala Tyr Cys Thr Thr Gln Val 
    50                  55                  60                  


Glu Arg Asn Ala Cys Leu Phe Cys Lys Ala Arg Lys Leu Asp Asp Lys 
65                  70                  75                  80  


Phe Tyr Gln Lys Leu Arg Gly Gln Phe Pro Asp Ala Val Phe Trp Gln 
                85                  90                  95      


Glu Ile Ser Glu Ile Phe Arg Gln Leu Gln Lys Gln Ala Ala Glu Ile 
            100                 105                 110         


Tyr Asn Gln Ser Leu Ile Glu Leu Tyr Tyr Glu Ile Phe Ile Lys Gly 
        115                 120                 125             


Lys Gly Ile Ala Asn Ala Ser Ser Val Glu His Tyr Leu Ser Asp Val 
    130                 135                 140                 


Cys Tyr Thr Arg Ala Ala Glu Leu Phe Lys Asn Ala Ala Ile Ala Ser 
145                 150                 155                 160 


Gly Leu Arg Ser Lys Ile Lys Ser Asn Phe Arg Leu Lys Glu Leu Lys 
                165                 170                 175     


Asn Met Lys Ser Gly Leu Pro Thr Thr Lys Ser Asp Asn Phe Pro Ile 
            180                 185                 190         


Pro Leu Val Lys Gln Lys Gly Gly Gln Tyr Thr Gly Phe Glu Ile Ser 
        195                 200                 205             


Asn His Asn Ser Asp Phe Ile Ile Lys Ile Pro Phe Gly Arg Trp Gln 
    210                 215                 220                 


Val Lys Lys Glu Ile Asp Lys Tyr Arg Pro Trp Glu Lys Phe Asp Phe 
225                 230                 235                 240 


Glu Gln Val Gln Lys Ser Pro Lys Pro Ile Ser Leu Leu Leu Ser Thr 
                245                 250                 255     


Gln Arg Arg Lys Arg Asn Lys Gly Trp Ser Lys Asp Glu Gly Thr Glu 
            260                 265                 270         


Ala Glu Ile Lys Lys Val Met Asn Gly Asp Tyr Gln Thr Ser Tyr Ile 
        275                 280                 285             


Glu Val Lys Arg Gly Ser Lys Ile Gly Glu Lys Ser Ala Trp Met Leu 
    290                 295                 300                 


Asn Leu Ser Ile Asp Val Pro Lys Ile Asp Lys Gly Val Asp Pro Ser 
305                 310                 315                 320 


Ile Ile Gly Gly Ile Asp Val Gly Val Lys Ser Pro Leu Val Cys Ala 
                325                 330                 335     


Ile Asn Asn Ala Phe Ser Arg Tyr Ser Ile Ser Asp Asn Asp Leu Phe 
            340                 345                 350         


His Phe Asn Lys Lys Met Phe Ala Arg Arg Arg Ile Leu Leu Lys Lys 
        355                 360                 365             


Asn Arg His Lys Arg Ala Gly His Gly Ala Lys Asn Lys Leu Lys Pro 
    370                 375                 380                 


Ile Thr Ile Leu Thr Glu Lys Ser Glu Arg Phe Arg Lys Lys Leu Ile 
385                 390                 395                 400 


Glu Arg Trp Ala Cys Glu Ile Ala Asp Phe Phe Ile Lys Asn Lys Val 
                405                 410                 415     


Gly Thr Val Gln Met Glu Asn Leu Glu Ser Met Lys Arg Lys Glu Asp 
            420                 425                 430         


Ser Tyr Phe Asn Ile Arg Leu Arg Gly Phe Trp Pro Tyr Ala Glu Met 
        435                 440                 445             


Gln Asn Lys Ile Glu Phe Lys Leu Lys Gln Tyr Gly Ile Glu Ile Arg 
    450                 455                 460                 


Lys Val Ala Pro Asn Asn Thr Ser Lys Thr Cys Ser Lys Cys Gly His 
465                 470                 475                 480 


Leu Asn Asn Tyr Phe Asn Phe Glu Tyr Arg Lys Lys Asn Lys Phe Pro 
                485                 490                 495     


His Phe Lys Cys Glu Lys Cys Asn Phe Lys Glu Asn Ala Asp Tyr Asn 
            500                 505                 510         


Ala Ala Leu Asn Ile Ser Asn Pro Lys Leu Lys Ser Thr Lys Glu Glu 
        515                 520                 525             


Pro 
    


<210>  309
<211>  564
<212>  PRT
<213>  Uncultured archaeon


<220>
<221>  MISC_FEATURE
<223>  Cas14b5

<400>  309

Met Ala Arg Ala Lys Asn Gln Pro Tyr Gln Lys Leu Thr Thr Thr Thr 
1               5                   10                  15      


Gly Ile Lys Phe Lys Leu Asp Leu Ser Glu Glu Glu Gly Lys Arg Phe 
            20                  25                  30          


Asp Glu Tyr Phe Ser Glu Tyr Ala Lys Ala Val Asn Phe Cys Ala Lys 
        35                  40                  45              


Val Ile Tyr Gln Leu Arg Lys Asn Leu Lys Phe Ala Gly Lys Lys Glu 
    50                  55                  60                  


Leu Ala Ala Lys Glu Trp Lys Phe Glu Ile Ser Asn Cys Asp Phe Cys 
65                  70                  75                  80  


Asn Lys Gln Lys Glu Ile Tyr Tyr Lys Asn Ile Ala Asn Gly Gln Lys 
                85                  90                  95      


Val Cys Lys Gly Cys His Arg Thr Asn Phe Ser Asp Asn Ala Ile Arg 
            100                 105                 110         


Lys Lys Met Ile Pro Val Lys Gly Arg Lys Val Glu Ser Lys Phe Asn 
        115                 120                 125             


Ile His Asn Thr Thr Lys Lys Ile Ser Gly Thr His Arg His Trp Ala 
    130                 135                 140                 


Phe Glu Asp Ala Ala Asp Ile Ile Glu Ser Met Asp Lys Gln Arg Lys 
145                 150                 155                 160 


Glu Lys Gln Lys Arg Leu Arg Arg Glu Lys Arg Lys Leu Ser Tyr Phe 
                165                 170                 175     


Phe Glu Leu Phe Gly Asp Pro Ala Lys Arg Tyr Glu Leu Pro Lys Val 
            180                 185                 190         


Gly Lys Gln Arg Val Pro Arg Tyr Leu His Lys Ile Ile Asp Lys Asp 
        195                 200                 205             


Ser Leu Thr Lys Lys Arg Gly Tyr Ser Leu Ser Tyr Ile Lys Asn Lys 
    210                 215                 220                 


Ile Lys Ile Ser Glu Arg Asn Ile Glu Arg Asp Glu Lys Ser Leu Arg 
225                 230                 235                 240 


Lys Ala Ser Pro Ile Ala Phe Gly Ala Arg Lys Ile Lys Met Ser Lys 
                245                 250                 255     


Leu Asp Pro Lys Arg Ala Phe Asp Leu Glu Asn Asn Val Phe Lys Ile 
            260                 265                 270         


Pro Gly Lys Val Ile Lys Gly Gln Tyr Lys Phe Phe Gly Thr Asn Val 
        275                 280                 285             


Ala Asn Glu His Gly Lys Lys Phe Tyr Lys Asp Arg Ile Ser Lys Ile 
    290                 295                 300                 


Leu Ala Gly Lys Pro Lys Tyr Phe Tyr Leu Leu Arg Lys Lys Val Ala 
305                 310                 315                 320 


Glu Ser Asp Gly Asn Pro Ile Phe Glu Tyr Tyr Val Gln Trp Ser Ile 
                325                 330                 335     


Asp Thr Glu Thr Pro Ala Ile Thr Ser Tyr Asp Asn Ile Leu Gly Ile 
            340                 345                 350         


Asp Ala Gly Ile Thr Asn Leu Ala Thr Thr Val Leu Ile Pro Lys Asn 
        355                 360                 365             


Leu Ser Ala Glu His Cys Ser His Cys Gly Asn Asn His Val Lys Pro 
    370                 375                 380                 


Ile Phe Thr Lys Phe Phe Ser Gly Lys Glu Leu Lys Ala Ile Lys Ile 
385                 390                 395                 400 


Lys Ser Arg Lys Gln Lys Tyr Phe Leu Arg Gly Lys His Asn Lys Leu 
                405                 410                 415     


Val Lys Ile Lys Arg Ile Arg Pro Ile Glu Gln Lys Val Asp Gly Tyr 
            420                 425                 430         


Cys His Val Val Ser Lys Gln Ile Val Glu Met Ala Lys Glu Arg Asn 
        435                 440                 445             


Ser Cys Ile Ala Leu Glu Lys Leu Glu Lys Pro Lys Lys Ser Lys Phe 
    450                 455                 460                 


Arg Gln Arg Arg Arg Glu Lys Tyr Ala Val Ser Met Phe Val Phe Lys 
465                 470                 475                 480 


Lys Leu Ala Thr Phe Ile Lys Tyr Lys Ala Ala Arg Glu Gly Ile Glu 
                485                 490                 495     


Ile Ile Pro Val Glu Pro Glu Gly Thr Ser Tyr Thr Cys Ser His Cys 
            500                 505                 510         


Lys Asn Ala Gln Asn Asn Gln Arg Pro Tyr Phe Lys Pro Asn Ser Lys 
        515                 520                 525             


Lys Ser Trp Thr Ser Met Phe Lys Cys Gly Lys Cys Gly Ile Glu Leu 
    530                 535                 540                 


Asn Ser Asp Tyr Asn Ala Ala Phe Asn Ile Ala Gln Lys Ala Leu Asn 
545                 550                 555                 560 


Met Thr Ser Ala 
                


<210>  310
<211>  1300
<212>  PRT
<213>  Francisella tularensis


<220>
<221>  MISC_FEATURE
<223>  Cas12a

<400>  310

Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr 
1               5                   10                  15      


Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys 
            20                  25                  30          


Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys 
        35                  40                  45              


Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu 
    50                  55                  60                  


Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser 
65                  70                  75                  80  


Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys 
                85                  90                  95      


Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr 
            100                 105                 110         


Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile 
        115                 120                 125             


Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln 
    130                 135                 140                 


Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr 
145                 150                 155                 160 


Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr 
                165                 170                 175     


Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser 
            180                 185                 190         


Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu 
        195                 200                 205             


Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys 
    210                 215                 220                 


Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu 
225                 230                 235                 240 


Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg 
                245                 250                 255     


Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr 
            260                 265                 270         


Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys 
        275                 280                 285             


Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile 
    290                 295                 300                 


Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys 
305                 310                 315                 320 


Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser 
                325                 330                 335     


Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met 
            340                 345                 350         


Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys 
        355                 360                 365             


Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln 
    370                 375                 380                 


Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr 
385                 390                 395                 400 


Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala 
                405                 410                 415     


Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn 
            420                 425                 430         


Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala 
        435                 440                 445             


Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn 
    450                 455                 460                 


Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala 
465                 470                 475                 480 


Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys 
                485                 490                 495     


Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys 
            500                 505                 510         


Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp 
        515                 520                 525             


Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His 
    530                 535                 540                 


Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His 
545                 550                 555                 560 


Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val 
                565                 570                 575     


Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser 
            580                 585                 590         


Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly 
        595                 600                 605             


Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys 
    610                 615                 620                 


Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile 
625                 630                 635                 640 


Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys 
                645                 650                 655     


Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val 
            660                 665                 670         


Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile 
        675                 680                 685             


Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln 
    690                 695                 700                 


Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe 
705                 710                 715                 720 


Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp 
                725                 730                 735     


Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu 
            740                 745                 750         


Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn 
        755                 760                 765             


Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr 
    770                 775                 780                 


Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg 
785                 790                 795                 800 


Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn 
                805                 810                 815     


Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr 
            820                 825                 830         


Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala 
        835                 840                 845             


Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu 
    850                 855                 860                 


Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe 
865                 870                 875                 880 


His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe 
                885                 890                 895     


Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His 
            900                 905                 910         


Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu 
        915                 920                 925             


Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile 
    930                 935                 940                 


Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile 
945                 950                 955                 960 


Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn 
                965                 970                 975     


Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile 
            980                 985                 990         


Ala Lys Leu Val Ile Glu Tyr Asn  Ala Ile Val Val Phe  Glu Asp Leu 
        995                 1000                 1005             


Asn Phe  Gly Phe Lys Arg Gly  Arg Phe Lys Val Glu  Lys Gln Val 
    1010                 1015                 1020             


Tyr Gln  Lys Leu Glu Lys Met  Leu Ile Glu Lys Leu  Asn Tyr Leu 
    1025                 1030                 1035             


Val Phe  Lys Asp Asn Glu Phe  Asp Lys Thr Gly Gly  Val Leu Arg 
    1040                 1045                 1050             


Ala Tyr  Gln Leu Thr Ala Pro  Phe Glu Thr Phe Lys  Lys Met Gly 
    1055                 1060                 1065             


Lys Gln  Thr Gly Ile Ile Tyr  Tyr Val Pro Ala Gly  Phe Thr Ser 
    1070                 1075                 1080             


Lys Ile  Cys Pro Val Thr Gly  Phe Val Asn Gln Leu  Tyr Pro Lys 
    1085                 1090                 1095             


Tyr Glu  Ser Val Ser Lys Ser  Gln Glu Phe Phe Ser  Lys Phe Asp 
    1100                 1105                 1110             


Lys Ile  Cys Tyr Asn Leu Asp  Lys Gly Tyr Phe Glu  Phe Ser Phe 
    1115                 1120                 1125             


Asp Tyr  Lys Asn Phe Gly Asp  Lys Ala Ala Lys Gly  Lys Trp Thr 
    1130                 1135                 1140             


Ile Ala  Ser Phe Gly Ser Arg  Leu Ile Asn Phe Arg  Asn Ser Asp 
    1145                 1150                 1155             


Lys Asn  His Asn Trp Asp Thr  Arg Glu Val Tyr Pro  Thr Lys Glu 
    1160                 1165                 1170             


Leu Glu  Lys Leu Leu Lys Asp  Tyr Ser Ile Glu Tyr  Gly His Gly 
    1175                 1180                 1185             


Glu Cys  Ile Lys Ala Ala Ile  Cys Gly Glu Ser Asp  Lys Lys Phe 
    1190                 1195                 1200             


Phe Ala  Lys Leu Thr Ser Val  Leu Asn Thr Ile Leu  Gln Met Arg 
    1205                 1210                 1215             


Asn Ser  Lys Thr Gly Thr Glu  Leu Asp Tyr Leu Ile  Ser Pro Val 
    1220                 1225                 1230             


Ala Asp  Val Asn Gly Asn Phe  Phe Asp Ser Arg Gln  Ala Pro Lys 
    1235                 1240                 1245             


Asn Met  Pro Gln Asp Ala Asp  Ala Asn Gly Ala Tyr  His Ile Gly 
    1250                 1255                 1260             


Leu Lys  Gly Leu Met Leu Leu  Gly Arg Ile Lys Asn  Asn Gln Glu 
    1265                 1270                 1275             


Gly Lys  Lys Leu Asn Leu Val  Ile Lys Asn Glu Glu  Tyr Phe Glu 
    1280                 1285                 1290             


Phe Val  Gln Asn Arg Asn Asn  
    1295                 1300 


<210>  311
<211>  1307
<212>  PRT
<213>  Acidaminococcus sp

<400>  311

Met Thr Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr 
1               5                   10                  15      


Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln 
            20                  25                  30          


Glu Gln Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys 
        35                  40                  45              


Glu Leu Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln 
    50                  55                  60                  


Cys Leu Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile 
65                  70                  75                  80  


Asp Ser Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile 
                85                  90                  95      


Glu Glu Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly 
            100                 105                 110         


Arg Thr Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile 
        115                 120                 125             


Tyr Lys Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys 
    130                 135                 140                 


Gln Leu Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg 
145                 150                 155                 160 


Ser Phe Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg 
                165                 170                 175     


Lys Asn Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg 
            180                 185                 190         


Ile Val Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe 
        195                 200                 205             


Thr Arg Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn 
    210                 215                 220                 


Val Lys Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val 
225                 230                 235                 240 


Phe Ser Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp 
                245                 250                 255     


Leu Tyr Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu 
            260                 265                 270         


Lys Ile Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn 
        275                 280                 285             


Asp Glu Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro 
    290                 295                 300                 


Leu Phe Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu 
305                 310                 315                 320 


Glu Glu Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr 
                325                 330                 335     


Lys Thr Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu 
            340                 345                 350         


Phe Asn Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His 
        355                 360                 365             


Lys Lys Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr 
    370                 375                 380                 


Leu Arg Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys 
385                 390                 395                 400 


Ile Thr Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu 
                405                 410                 415     


Asp Ile Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser 
            420                 425                 430         


Glu Ala Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala 
        435                 440                 445             


Ala Leu Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys 
    450                 455                 460                 


Glu Ile Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu 
465                 470                 475                 480 


Leu Asp Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe 
                485                 490                 495     


Ser Ala Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser 
            500                 505                 510         


Phe Tyr Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val 
        515                 520                 525             


Glu Lys Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp 
    530                 535                 540                 


Asp Val Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn 
545                 550                 555                 560 


Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr Lys 
                565                 570                 575     


Ala Leu Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys 
            580                 585                 590         


Met Tyr Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys 
        595                 600                 605             


Ser Thr Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr 
    610                 615                 620                 


Pro Ile Leu Leu Ser Asn Asn Phe Ile Glu Pro Leu Glu Ile Thr Lys 
625                 630                 635                 640 


Glu Ile Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln 
                645                 650                 655     


Thr Ala Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala 
            660                 665                 670         


Leu Cys Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr 
        675                 680                 685             


Lys Thr Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser Gln Tyr 
    690                 695                 700                 


Lys Asp Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His 
705                 710                 715                 720 


Ile Ser Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu 
                725                 730                 735     


Thr Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys 
            740                 745                 750         


Gly His His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu 
        755                 760                 765             


Phe Ser Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln 
    770                 775                 780                 


Ala Glu Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His 
785                 790                 795                 800 


Arg Leu Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr 
                805                 810                 815     


Pro Ile Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His 
            820                 825                 830         


Arg Leu Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn 
        835                 840                 845             


Val Ile Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe 
    850                 855                 860                 


Thr Ser Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln 
865                 870                 875                 880 


Ala Ala Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu 
                885                 890                 895     


Lys Glu His Pro Glu Thr Pro Ile Ile Gly Ile Asp Arg Gly Glu Arg 
            900                 905                 910         


Asn Leu Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu 
        915                 920                 925             


Gln Arg Ser Leu Asn Thr Ile Gln Gln Phe Asp Tyr Gln Lys Lys Leu 
    930                 935                 940                 


Asp Asn Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val 
945                 950                 955                 960 


Val Gly Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile 
                965                 970                 975     


His Glu Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu 
            980                 985                 990         


Glu Asn Leu Asn Phe Gly Phe Lys  Ser Lys Arg Thr Gly  Ile Ala Glu 
        995                 1000                 1005             


Lys Ala  Val Tyr Gln Gln Phe  Glu Lys Met Leu Ile  Asp Lys Leu 
    1010                 1015                 1020             


Asn Cys  Leu Val Leu Lys Asp  Tyr Pro Ala Glu Lys  Val Gly Gly 
    1025                 1030                 1035             


Val Leu  Asn Pro Tyr Gln Leu  Thr Asp Gln Phe Thr  Ser Phe Ala 
    1040                 1045                 1050             


Lys Met  Gly Thr Gln Ser Gly  Phe Leu Phe Tyr Val  Pro Ala Pro 
    1055                 1060                 1065             


Tyr Thr  Ser Lys Ile Asp Pro  Leu Thr Gly Phe Val  Asp Pro Phe 
    1070                 1075                 1080             


Val Trp  Lys Thr Ile Lys Asn  His Glu Ser Arg Lys  His Phe Leu 
    1085                 1090                 1095             


Glu Gly  Phe Asp Phe Leu His  Tyr Asp Val Lys Thr  Gly Asp Phe 
    1100                 1105                 1110             


Ile Leu  His Phe Lys Met Asn  Arg Asn Leu Ser Phe  Gln Arg Gly 
    1115                 1120                 1125             


Leu Pro  Gly Phe Met Pro Ala  Trp Asp Ile Val Phe  Glu Lys Asn 
    1130                 1135                 1140             


Glu Thr  Gln Phe Asp Ala Lys  Gly Thr Pro Phe Ile  Ala Gly Lys 
    1145                 1150                 1155             


Arg Ile  Val Pro Val Ile Glu  Asn His Arg Phe Thr  Gly Arg Tyr 
    1160                 1165                 1170             


Arg Asp  Leu Tyr Pro Ala Asn  Glu Leu Ile Ala Leu  Leu Glu Glu 
    1175                 1180                 1185             


Lys Gly  Ile Val Phe Arg Asp  Gly Ser Asn Ile Leu  Pro Lys Leu 
    1190                 1195                 1200             


Leu Glu  Asn Asp Asp Ser His  Ala Ile Asp Thr Met  Val Ala Leu 
    1205                 1210                 1215             


Ile Arg  Ser Val Leu Gln Met  Arg Asn Ser Asn Ala  Ala Thr Gly 
    1220                 1225                 1230             


Glu Asp  Tyr Ile Asn Ser Pro  Val Arg Asp Leu Asn  Gly Val Cys 
    1235                 1240                 1245             


Phe Asp  Ser Arg Phe Gln Asn  Pro Glu Trp Pro Met  Asp Ala Asp 
    1250                 1255                 1260             


Ala Asn  Gly Ala Tyr His Ile  Ala Leu Lys Gly Gln  Leu Leu Leu 
    1265                 1270                 1275             


Asn His  Leu Lys Glu Ser Lys  Asp Leu Lys Leu Gln  Asn Gly Ile 
    1280                 1285                 1290             


Ser Asn  Gln Asp Trp Leu Ala  Tyr Ile Gln Glu Leu  Arg Asn 
    1295                 1300                 1305         


<210>  312
<211>  1228
<212>  PRT
<213>  Lachnospiraceae bacterium

<400>  312

Ala Ala Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys 
1               5                   10                  15      


Thr Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile 
            20                  25                  30          


Asp Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr 
        35                  40                  45              


Lys Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn 
    50                  55                  60                  


Asp Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser 
65                  70                  75                  80  


Leu Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu 
                85                  90                  95      


Asn Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly 
            100                 105                 110         


Ala Ala Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile 
        115                 120                 125             


Leu Pro Glu Ala Ala Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser 
    130                 135                 140                 


Phe Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu 
145                 150                 155                 160 


Asn Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys 
                165                 170                 175     


Ile Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu 
            180                 185                 190         


Lys Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu 
        195                 200                 205             


Lys Ile Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu 
    210                 215                 220                 


Phe Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala 
225                 230                 235                 240 


Ile Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu 
                245                 250                 255     


Asn Glu Tyr Ile Asn Leu Tyr Asn Ala Lys Thr Lys Gln Ala Leu Pro 
            260                 265                 270         


Lys Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu 
        275                 280                 285             


Ser Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val 
    290                 295                 300                 


Phe Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys 
305                 310                 315                 320 


Lys Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly 
                325                 330                 335     


Ile Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile 
            340                 345                 350         


Phe Gly Glu Trp Asn Leu Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp 
        355                 360                 365             


Asp Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp 
    370                 375                 380                 


Asp Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln 
385                 390                 395                 400 


Leu Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys 
                405                 410                 415     


Glu Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser 
            420                 425                 430         


Ser Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys 
        435                 440                 445             


Lys Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val 
    450                 455                 460                 


Lys Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu 
465                 470                 475                 480 


Thr Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp 
                485                 490                 495     


Ile Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val 
            500                 505                 510         


Thr Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn 
        515                 520                 525             


Pro Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg 
    530                 535                 540                 


Ala Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp 
545                 550                 555                 560 


Lys Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn 
                565                 570                 575     


Gly Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys 
            580                 585                 590         


Met Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn 
        595                 600                 605             


Pro Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys 
    610                 615                 620                 


Gly Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe 
625                 630                 635                 640 


Lys Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe 
                645                 650                 655     


Asn Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg 
            660                 665                 670         


Glu Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys 
        675                 680                 685             


Lys Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln 
    690                 695                 700                 


Ile Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu 
705                 710                 715                 720 


His Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln 
                725                 730                 735     


Ile Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu 
            740                 745                 750         


Lys Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn 
        755                 760                 765             


Lys Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val 
    770                 775                 780                 


Tyr Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro 
785                 790                 795                 800 


Ile Ala Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu 
                805                 810                 815     


Val Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile 
            820                 825                 830         


Asp Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys 
        835                 840                 845             


Gly Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe 
    850                 855                 860                 


Asn Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys 
865                 870                 875                 880 


Glu Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn 
                885                 890                 895     


Ile Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile 
            900                 905                 910         


Cys Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 
        915                 920                 925             


Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr 
    930                 935                 940                 


Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp 
945                 950                 955                 960 


Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln 
                965                 970                 975     


Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly 
            980                 985                 990         


Phe Ile Phe Tyr Ile Pro Ala Trp  Leu Thr Ser Lys Ile  Asp Pro Ser 
        995                 1000                 1005             


Thr Gly  Phe Val Asn Leu Leu  Lys Thr Lys Tyr Thr  Ser Ile Ala 
    1010                 1015                 1020             


Asp Ser  Lys Lys Phe Ile Ser  Ser Phe Asp Arg Ile  Met Tyr Val 
    1025                 1030                 1035             


Pro Glu  Glu Asp Leu Phe Glu  Phe Ala Leu Asp Tyr  Lys Asn Phe 
    1040                 1045                 1050             


Ser Arg  Thr Asp Ala Asp Tyr  Ile Lys Lys Trp Lys  Leu Tyr Ser 
    1055                 1060                 1065             


Tyr Gly  Asn Arg Ile Arg Ile  Phe Ala Ala Ala Lys  Lys Asn Asn 
    1070                 1075                 1080             


Val Phe  Ala Trp Glu Glu Val  Cys Leu Thr Ser Ala  Tyr Lys Glu 
    1085                 1090                 1095             


Leu Phe  Asn Lys Tyr Gly Ile  Asn Tyr Gln Gln Gly  Asp Ile Arg 
    1100                 1105                 1110             


Ala Leu  Leu Cys Glu Gln Ser  Asp Lys Ala Phe Tyr  Ser Ser Phe 
    1115                 1120                 1125             


Met Ala  Leu Met Ser Leu Met  Leu Gln Met Arg Asn  Ser Ile Thr 
    1130                 1135                 1140             


Gly Arg  Thr Asp Val Asp Phe  Leu Ile Ser Pro Val  Lys Asn Ser 
    1145                 1150                 1155             


Asp Gly  Ile Phe Tyr Asp Ser  Arg Asn Tyr Glu Ala  Gln Glu Asn 
    1160                 1165                 1170             


Ala Ile  Leu Pro Lys Asn Ala  Asp Ala Asn Gly Ala  Tyr Asn Ile 
    1175                 1180                 1185             


Ala Arg  Lys Val Leu Trp Ala  Ile Gly Gln Phe Lys  Lys Ala Glu 
    1190                 1195                 1200             


Asp Glu  Lys Leu Asp Lys Val  Lys Ile Ala Ile Ser  Asn Lys Glu 
    1205                 1210                 1215             


Trp Leu  Glu Tyr Ala Gln Thr  Ser Val Lys 
    1220                 1225             


<210>  313
<211>  1053
<212>  PRT
<213>  Streptococcus aureus

<400>  313

Met Lys Arg Asn Tyr Ile Leu Gly Leu Asp Ile Gly Ile Thr Ser Val 
1               5                   10                  15      


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


Gly Lys Glu Asn Ala Lys Tyr Leu Ile Glu Lys Ile Lys Leu His Asp 
        515                 520                 525             


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


Val Lys Ser Ile Asn Gly Gly Phe Thr Ser Phe Leu Arg Arg Lys Trp 
        675                 680                 685             


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Glu Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Asn Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro Arg  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  314
<211>  1526
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, HNHdelta, Zinc finger PAMBD loop replacement, 2NLS

<400>  314

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1340                 1345                 1350             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1355                 1360                 1365             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1370                 1375                 1380             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1385                 1390                 1395             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1400                 1405                 1410             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1415                 1420                 1425             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly Ser Ala  Gly Gly Thr 
    1430                 1435                 1440             


Gly Gly  Ser Ala Pro Tyr Arg  Cys Pro Cys Arg Ala  Gly Cys Pro 
    1445                 1450                 1455             


Ser Leu  Ala Ser Met Gln Ala  His Met Arg Gly His  Ser Gly Gly 
    1460                 1465                 1470             


Ser Gly  Tyr Gln Thr Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile 
    1475                 1480                 1485             


Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser 
    1490                 1495                 1500             


Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly 
    1505                 1510                 1515             


Gln Ala  Lys Lys Lys Lys Gly  Ala 
    1520                 1525     


<210>  315
<211>  1531
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, HNHdelta, Zinc finger PAMBD loop replacement longer
       linkers, 2NLS

<400>  315

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1340                 1345                 1350             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1355                 1360                 1365             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1370                 1375                 1380             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1385                 1390                 1395             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1400                 1405                 1410             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1415                 1420                 1425             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly Ser Ala  Gly Gly Thr 
    1430                 1435                 1440             


Gly Gly  Ser Ala Gly Gly Ser  Gly Ala Pro Tyr Arg  Cys Pro Cys 
    1445                 1450                 1455             


Arg Ala  Gly Cys Pro Ser Leu  Ala Ser Met Gln Ala  His Met Arg 
    1460                 1465                 1470             


Gly His  Ser Gly Gly Ser Gly  Tyr Gln Thr Val Thr  Glu Val Leu 
    1475                 1480                 1485             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1490                 1495                 1500             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala 
    1505                 1510                 1515             


Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys Lys Gly  Ala 
    1520                 1525                 1530     


<210>  316
<211>  1446
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, HNHdelta, Lac repressor DBD PAMBD replacement whole
       replacement, 2NLS

<400>  316

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Gly Gly  Ser Gly Met Lys Pro  Val Thr Leu 
    1340                 1345                 1350             


Tyr Asp  Val Ala Glu Tyr Ala  Gly Val Ser Tyr Gln  Thr Val Ser 
    1355                 1360                 1365             


Arg Val  Val Asn Gln Ala Ser  His Val Ser Ala Lys  Thr Arg Glu 
    1370                 1375                 1380             


Lys Val  Glu Ala Ala Met Ala  Glu Leu Asn Gly Gly  Ser Gly Tyr 
    1385                 1390                 1395             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1400                 1405                 1410             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1415                 1420                 1425             


Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys 
    1430                 1435                 1440             


Lys Lys  Lys 
    1445     


<210>  317
<211>  1466
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, HNHdelta, SSO7D PAMBD replacement whole 
       replacement, 2NLS

<400>  317

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly Met Ala  Thr Val Lys 
    1340                 1345                 1350             


Phe Lys  Tyr Lys Gly Glu Glu  Lys Glu Val Asp Ile  Ser Lys Ile 
    1355                 1360                 1365             


Lys Lys  Val Trp Arg Val Gly  Lys Met Ile Ser Phe  Thr Tyr Asp 
    1370                 1375                 1380             


Glu Gly  Gly Gly Lys Thr Gly  Arg Gly Ala Val Ser  Glu Lys Asp 
    1385                 1390                 1395             


Ala Pro  Lys Glu Leu Leu Gln  Met Leu Glu Lys Gln  Lys Lys Gly 
    1400                 1405                 1410             


Gly Ser  Ala Gly Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1415                 1420                 1425             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1430                 1435                 1440             


Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys 
    1445                 1450                 1455             


Ala Gly  Gln Ala Lys Lys Lys  Lys 
    1460                 1465     


<210>  318
<211>  1571
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, HNHdelta, STO7D PAMBD replacement loop replacement 
       longer linker, 2NLS

<400>  318

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1340                 1345                 1350             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1355                 1360                 1365             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1370                 1375                 1380             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1385                 1390                 1395             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1400                 1405                 1410             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1415                 1420                 1425             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly Ser Ala  Gly Gly Thr 
    1430                 1435                 1440             


Gly Gly  Ser Met Val Thr Val  Lys Phe Lys Tyr Lys  Gly Glu Glu 
    1445                 1450                 1455             


Lys Glu  Val Asp Ile Ser Lys  Ile Lys Lys Val Trp  Arg Val Gly 
    1460                 1465                 1470             


Lys Met  Ile Ser Phe Thr Tyr  Asp Asp Asn Gly Lys  Thr Gly Arg 
    1475                 1480                 1485             


Gly Ala  Val Ser Glu Lys Asp  Ala Pro Lys Glu Leu  Leu Gln Met 
    1490                 1495                 1500             


Leu Glu  Lys Ser Gly Lys Lys  Gly Gly Ser Ala Gly  Gly Thr Gly 
    1505                 1510                 1515             


Gly Ser  Tyr Gln Thr Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile 
    1520                 1525                 1530             


Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser 
    1535                 1540                 1545             


Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly 
    1550                 1555                 1560             


Gln Ala  Lys Lys Lys Lys Gly  Ala 
    1565                 1570     


<210>  319
<211>  593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  MPO.ex1.dcasffok.pamout

<400>  319
acgcgtaaaa aaaaactggc cacgccccag ccctcgtccc ctcgtgaggg gcaatcgttg       60

ggtgtttcgt ggtctcatac agaacttata agattcccaa atccaaagac atttcacgtt      120

tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagggcgcc actcccctgt      180

ccctcacagc catcttcctg ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac      240

tgggcccgcg attccttgga gcgggttgat gacgtcagcg ttcaaagagg gcctatttcc      300

catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa      360

tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct      420

tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac      480

ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacccaacg      540

attgcccctc acgaggggac agctgcttct ggccctagca tttttttacg cgt             593


<210>  320
<211>  1684
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas-FokI-HMGB1

<400>  320

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
                805                 810                 815     


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
            820                 825                 830         


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
        835                 840                 845             


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
    850                 855                 860                 


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
865                 870                 875                 880 


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
                885                 890                 895     


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
            900                 905                 910         


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
        915                 920                 925             


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
    930                 935                 940                 


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
945                 950                 955                 960 


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
                965                 970                 975     


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
            980                 985                 990         


Asn Ile Val Ile Glu Met Ala Arg  Glu Asn Gln Thr Thr  Thr Lys Gly 
        995                 1000                 1005             


Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg Ile Glu  Glu Gly Ile 
    1010                 1015                 1020             


Lys Glu  Leu Glu Ser Gln Ile  Leu Lys Glu Asn Pro  Val Glu Asn 
    1025                 1030                 1035             


Thr Gln  Leu Gln Asn Glu Lys  Leu Tyr Leu Tyr Tyr  Leu Gln Asn 
    1040                 1045                 1050             


Gly Arg  Asp Met Tyr Val Asp  Gln Glu Leu Asp Ile  Asn Arg Leu 
    1055                 1060                 1065             


Ser Asp  Tyr Asp Val Asp Ala  Ile Val Pro Gln Ser  Phe Ile Lys 
    1070                 1075                 1080             


Asp Asp  Ser Ile Asp Asn Lys  Val Leu Thr Arg Ser  Val Glu Asn 
    1085                 1090                 1095             


Arg Gly  Lys Ser Asp Asn Val  Pro Ser Glu Glu Val  Val Lys Lys 
    1100                 1105                 1110             


Met Lys  Asn Tyr Trp Arg Gln  Leu Leu Asn Ala Lys  Leu Ile Thr 
    1115                 1120                 1125             


Gln Arg  Lys Phe Asp Asn Leu  Thr Lys Ala Glu Arg  Gly Gly Leu 
    1130                 1135                 1140             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1145                 1150                 1155             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1160                 1165                 1170             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1175                 1180                 1185             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1190                 1195                 1200             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1205                 1210                 1215             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1220                 1225                 1230             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1235                 1240                 1245             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1250                 1255                 1260             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1265                 1270                 1275             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1280                 1285                 1290             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1295                 1300                 1305             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1310                 1315                 1320             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1325                 1330                 1335             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1340                 1345                 1350             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1355                 1360                 1365             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1370                 1375                 1380             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1385                 1390                 1395             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1400                 1405                 1410             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1415                 1420                 1425             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1430                 1435                 1440             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1445                 1450                 1455             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1460                 1465                 1470             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1475                 1480                 1485             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1490                 1495                 1500             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1505                 1510                 1515             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1520                 1525                 1530             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1535                 1540                 1545             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1550                 1555                 1560             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1565                 1570                 1575             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1580                 1585                 1590             


Asp Leu  Ser Gln Leu Gly Gly  Asp Gly Lys Gly Asp  Pro Lys Lys 
    1595                 1600                 1605             


Pro Arg  Gly Lys Met Ser Ser  Tyr Ala Phe Phe Val  Gln Thr Cys 
    1610                 1615                 1620             


Arg Glu  Glu His Lys Lys Lys  His Pro Asp Ala Ser  Val Asn Phe 
    1625                 1630                 1635             


Ser Glu  Phe Ser Lys Lys Cys  Ser Glu Arg Trp Lys  Thr Met Ser 
    1640                 1645                 1650             


Ala Lys  Glu Lys Gly Lys Phe  Glu Asp Met Ala Lys  Ala Asp Lys 
    1655                 1660                 1665             


Ala Arg  Tyr Glu Arg Glu Met  Lys Thr Tyr Ile Pro  Pro Lys Gly 
    1670                 1675                 1680             


Glu 
    


<210>  321
<211>  1687
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas-FokI-HMGB3

<400>  321

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
                805                 810                 815     


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
            820                 825                 830         


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
        835                 840                 845             


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
    850                 855                 860                 


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
865                 870                 875                 880 


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
                885                 890                 895     


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
            900                 905                 910         


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
        915                 920                 925             


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
    930                 935                 940                 


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
945                 950                 955                 960 


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
                965                 970                 975     


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
            980                 985                 990         


Asn Ile Val Ile Glu Met Ala Arg  Glu Asn Gln Thr Thr  Thr Lys Gly 
        995                 1000                 1005             


Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg Ile Glu  Glu Gly Ile 
    1010                 1015                 1020             


Lys Glu  Leu Glu Ser Gln Ile  Leu Lys Glu Asn Pro  Val Glu Asn 
    1025                 1030                 1035             


Thr Gln  Leu Gln Asn Glu Lys  Leu Tyr Leu Tyr Tyr  Leu Gln Asn 
    1040                 1045                 1050             


Gly Arg  Asp Met Tyr Val Asp  Gln Glu Leu Asp Ile  Asn Arg Leu 
    1055                 1060                 1065             


Ser Asp  Tyr Asp Val Asp Ala  Ile Val Pro Gln Ser  Phe Ile Lys 
    1070                 1075                 1080             


Asp Asp  Ser Ile Asp Asn Lys  Val Leu Thr Arg Ser  Val Glu Asn 
    1085                 1090                 1095             


Arg Gly  Lys Ser Asp Asn Val  Pro Ser Glu Glu Val  Val Lys Lys 
    1100                 1105                 1110             


Met Lys  Asn Tyr Trp Arg Gln  Leu Leu Asn Ala Lys  Leu Ile Thr 
    1115                 1120                 1125             


Gln Arg  Lys Phe Asp Asn Leu  Thr Lys Ala Glu Arg  Gly Gly Leu 
    1130                 1135                 1140             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1145                 1150                 1155             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1160                 1165                 1170             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1175                 1180                 1185             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1190                 1195                 1200             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1205                 1210                 1215             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1220                 1225                 1230             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1235                 1240                 1245             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1250                 1255                 1260             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1265                 1270                 1275             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1280                 1285                 1290             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1295                 1300                 1305             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1310                 1315                 1320             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1325                 1330                 1335             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1340                 1345                 1350             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1355                 1360                 1365             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1370                 1375                 1380             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1385                 1390                 1395             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1400                 1405                 1410             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1415                 1420                 1425             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1430                 1435                 1440             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1445                 1450                 1455             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1460                 1465                 1470             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1475                 1480                 1485             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1490                 1495                 1500             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1505                 1510                 1515             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1520                 1525                 1530             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1535                 1540                 1545             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1550                 1555                 1560             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1565                 1570                 1575             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1580                 1585                 1590             


Asp Leu  Ser Gln Leu Gly Gly  Asp Gly Ser Ser Gly  Ser Ser Gly 
    1595                 1600                 1605             


Met Ala  Lys Gly Asp Pro Lys  Lys Pro Lys Gly Lys  Met Ser Ala 
    1610                 1615                 1620             


Tyr Ala  Phe Phe Val Gln Thr  Cys Arg Glu Glu His  Lys Lys Lys 
    1625                 1630                 1635             


Asn Pro  Glu Val Pro Val Asn  Phe Ala Glu Phe Ser  Lys Lys Cys 
    1640                 1645                 1650             


Ser Glu  Arg Trp Lys Thr Met  Ser Gly Lys Glu Lys  Ser Lys Phe 
    1655                 1660                 1665             


Asp Glu  Met Ala Lys Ala Asp  Lys Val Arg Tyr Asp  Arg Glu Met 
    1670                 1675                 1680             


Lys Asp  Tyr Gly 
    1685         


<210>  322
<211>  1683
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas-FokI-HMGB4

<400>  322

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
                805                 810                 815     


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
            820                 825                 830         


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
        835                 840                 845             


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
    850                 855                 860                 


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
865                 870                 875                 880 


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
                885                 890                 895     


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
            900                 905                 910         


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
        915                 920                 925             


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
    930                 935                 940                 


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
945                 950                 955                 960 


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
                965                 970                 975     


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
            980                 985                 990         


Asn Ile Val Ile Glu Met Ala Arg  Glu Asn Gln Thr Thr  Thr Lys Gly 
        995                 1000                 1005             


Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg Ile Glu  Glu Gly Ile 
    1010                 1015                 1020             


Lys Glu  Leu Glu Ser Gln Ile  Leu Lys Glu Asn Pro  Val Glu Asn 
    1025                 1030                 1035             


Thr Gln  Leu Gln Asn Glu Lys  Leu Tyr Leu Tyr Tyr  Leu Gln Asn 
    1040                 1045                 1050             


Gly Arg  Asp Met Tyr Val Asp  Gln Glu Leu Asp Ile  Asn Arg Leu 
    1055                 1060                 1065             


Ser Asp  Tyr Asp Val Asp Ala  Ile Val Pro Gln Ser  Phe Ile Lys 
    1070                 1075                 1080             


Asp Asp  Ser Ile Asp Asn Lys  Val Leu Thr Arg Ser  Val Glu Asn 
    1085                 1090                 1095             


Arg Gly  Lys Ser Asp Asn Val  Pro Ser Glu Glu Val  Val Lys Lys 
    1100                 1105                 1110             


Met Lys  Asn Tyr Trp Arg Gln  Leu Leu Asn Ala Lys  Leu Ile Thr 
    1115                 1120                 1125             


Gln Arg  Lys Phe Asp Asn Leu  Thr Lys Ala Glu Arg  Gly Gly Leu 
    1130                 1135                 1140             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1145                 1150                 1155             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1160                 1165                 1170             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1175                 1180                 1185             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1190                 1195                 1200             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1205                 1210                 1215             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1220                 1225                 1230             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1235                 1240                 1245             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1250                 1255                 1260             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1265                 1270                 1275             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1280                 1285                 1290             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1295                 1300                 1305             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1310                 1315                 1320             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1325                 1330                 1335             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1340                 1345                 1350             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1355                 1360                 1365             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1370                 1375                 1380             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1385                 1390                 1395             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1400                 1405                 1410             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1415                 1420                 1425             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1430                 1435                 1440             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1445                 1450                 1455             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1460                 1465                 1470             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1475                 1480                 1485             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1490                 1495                 1500             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1505                 1510                 1515             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1520                 1525                 1530             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1535                 1540                 1545             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1550                 1555                 1560             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1565                 1570                 1575             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1580                 1585                 1590             


Asp Leu  Ser Gln Leu Gly Gly  Asp Gly Gly Ser Gly  Gly Lys Glu 
    1595                 1600                 1605             


Ile Gln  Leu Lys Pro Lys Ala  Asn Val Ser Ser Tyr  Val His Phe 
    1610                 1615                 1620             


Leu Leu  Asn Tyr Arg Asn Lys  Phe Lys Glu Gln Gln  Pro Asn Thr 
    1625                 1630                 1635             


Tyr Val  Gly Phe Lys Glu Phe  Ser Arg Lys Cys Ser  Glu Lys Trp 
    1640                 1645                 1650             


Arg Ser  Ile Ser Lys His Glu  Lys Ala Lys Tyr Glu  Ala Leu Ala 
    1655                 1660                 1665             


Lys Leu  Asp Lys Ala Arg Tyr  Gln Glu Glu Met Met  Asn Tyr Val 
    1670                 1675                 1680             


<210>  323
<211>  1714
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dScCasFok, 2NLS variant, Construct #13213

<400>  323

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Met Pro Lys Arg Lys 
1               5                   10                  15      


Val Ser Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser 
            20                  25                  30          


Ala Arg Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys 
        35                  40                  45              


Lys Ala Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys 
    50                  55                  60                  


Gly Lys Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu 
65                  70                  75                  80  


Thr Lys Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu 
                85                  90                  95      


Ser Pro Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Val 
            100                 105                 110         


Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys 
        115                 120                 125             


Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser 
    130                 135                 140                 


Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met Lys 
145                 150                 155                 160 


Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro Asp 
                165                 170                 175     


Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile Val 
            180                 185                 190         


Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala 
        195                 200                 205             


Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His 
    210                 215                 220                 


Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu 
225                 230                 235                 240 


Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys Ala 
                245                 250                 255     


Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val Leu 
            260                 265                 270         


Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly Thr 
        275                 280                 285             


Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile Asn 
    290                 295                 300                 


Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu 
305                 310                 315                 320 


Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser 
                325                 330                 335     


Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys 
            340                 345                 350         


Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn Leu 
        355                 360                 365             


Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg 
    370                 375                 380                 


Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile 
385                 390                 395                 400 


Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp Asp 
                405                 410                 415     


Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys 
            420                 425                 430         


Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val Ala 
        435                 440                 445             


Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala 
    450                 455                 460                 


Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala 
465                 470                 475                 480 


His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu Asn 
                485                 490                 495     


Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln Thr 
            500                 505                 510         


Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val Asp 
        515                 520                 525             


Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu Glu 
    530                 535                 540                 


Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe Gly 
545                 550                 555                 560 


Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn 
                565                 570                 575     


Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr 
            580                 585                 590         


Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr Ala 
        595                 600                 605             


Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser 
    610                 615                 620                 


Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser Ala 
625                 630                 635                 640 


Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu Leu 
                645                 650                 655     


Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile Phe 
            660                 665                 670         


Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly Ile 
        675                 680                 685             


Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe Tyr 
    690                 695                 700                 


Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu Leu 
705                 710                 715                 720 


Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe 
                725                 730                 735     


Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His Ala 
            740                 745                 750         


Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg 
        755                 760                 765             


Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly 
    770                 775                 780                 


Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser 
785                 790                 795                 800 


Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly 
                805                 810                 815     


Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu Gln 
            820                 825                 830         


Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr 
        835                 840                 845             


Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Arg 
    850                 855                 860                 


Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val 
865                 870                 875                 880 


Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys 
                885                 890                 895     


Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ile 
            900                 905                 910         


Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu 
        915                 920                 925             


Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu 
    930                 935                 940                 


Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg 
945                 950                 955                 960 


Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp 
                965                 970                 975     


Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly Arg 
            980                 985                 990         


Leu Ser Arg Lys Met Ile Asn Gly  Ile Arg Asp Lys Gln  Ser Gly Lys 
        995                 1000                 1005             


Thr Ile  Leu Asp Phe Leu Lys  Ser Asp Gly Phe Ser  Asn Arg Asn 
    1010                 1015                 1020             


Phe Met  Gln Leu Ile His Asp  Asp Ser Leu Thr Phe  Lys Glu Glu 
    1025                 1030                 1035             


Ile Glu  Lys Ala Gln Val Ser  Gly Gln Gly Asp Ser  Leu His Glu 
    1040                 1045                 1050             


Gln Ile  Ala Asp Leu Ala Gly  Ser Pro Ala Ile Lys  Lys Gly Ile 
    1055                 1060                 1065             


Leu Gln  Thr Val Lys Ile Val  Asp Glu Leu Val Lys  Val Met Gly 
    1070                 1075                 1080             


His Lys  Pro Glu Asn Ile Val  Ile Glu Met Ala Arg  Glu Asn Gln 
    1085                 1090                 1095             


Thr Thr  Thr Lys Gly Leu Gln  Gln Ser Arg Glu Arg  Lys Lys Arg 
    1100                 1105                 1110             


Ile Glu  Glu Gly Ile Lys Glu  Leu Glu Ser Gln Ile  Leu Lys Glu 
    1115                 1120                 1125             


Asn Pro  Val Glu Asn Thr Gln  Leu Gln Asn Glu Lys  Leu Tyr Leu 
    1130                 1135                 1140             


Tyr Tyr  Leu Gln Asn Gly Arg  Asp Met Tyr Val Asp  Gln Glu Leu 
    1145                 1150                 1155             


Asp Ile  Asn Arg Leu Ser Asp  Tyr Asp Val Asp Ala  Ile Val Pro 
    1160                 1165                 1170             


Gln Ser  Phe Ile Lys Asp Asp  Ser Ile Asp Asn Lys  Val Leu Thr 
    1175                 1180                 1185             


Arg Ser  Val Glu Asn Arg Gly  Lys Ser Asp Asn Val  Pro Ser Glu 
    1190                 1195                 1200             


Glu Val  Val Lys Lys Met Lys  Asn Tyr Trp Arg Gln  Leu Leu Asn 
    1205                 1210                 1215             


Ala Lys  Leu Ile Thr Gln Arg  Lys Phe Asp Asn Leu  Thr Lys Ala 
    1220                 1225                 1230             


Glu Arg  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1235                 1240                 1245             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1250                 1255                 1260             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1265                 1270                 1275             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1280                 1285                 1290             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1295                 1300                 1305             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1310                 1315                 1320             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1325                 1330                 1335             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1340                 1345                 1350             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1355                 1360                 1365             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1370                 1375                 1380             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1385                 1390                 1395             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1400                 1405                 1410             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1415                 1420                 1425             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1430                 1435                 1440             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1445                 1450                 1455             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1460                 1465                 1470             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1475                 1480                 1485             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1490                 1495                 1500             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1505                 1510                 1515             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1520                 1525                 1530             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1535                 1540                 1545             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1550                 1555                 1560             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1565                 1570                 1575             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1580                 1585                 1590             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1595                 1600                 1605             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1610                 1615                 1620             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1625                 1630                 1635             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu 
    1640                 1645                 1650             


Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr 
    1655                 1660                 1665             


Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu 
    1670                 1675                 1680             


Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg 
    1685                 1690                 1695             


Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala Lys Lys  Lys Lys Gly 
    1700                 1705                 1710             


Ala 
    


<210>  324
<211>  100
<212>  PRT
<213>  Homo sapiens


<220>
<221>  MISC_FEATURE
<223>  HMGN domain

<400>  324

Met Pro Lys Arg Lys Val Ser Ser Ala Glu Gly Ala Ala Lys Glu Glu 
1               5                   10                  15      


Pro Lys Arg Arg Ser Ala Arg Leu Ser Ala Lys Pro Pro Ala Lys Val 
            20                  25                  30          


Glu Ala Lys Pro Lys Lys Ala Ala Ala Lys Asp Lys Ser Ser Asp Lys 
        35                  40                  45              


Lys Val Gln Thr Lys Gly Lys Arg Gly Ala Lys Gly Lys Gln Ala Glu 
    50                  55                  60                  


Val Ala Asn Gln Glu Thr Lys Glu Asp Leu Pro Ala Glu Asn Gly Glu 
65                  70                  75                  80  


Thr Lys Thr Glu Glu Ser Pro Ala Ser Asp Glu Ala Gly Glu Lys Glu 
                85                  90                  95      


Ala Lys Ser Asp 
            100 


<210>  325
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GeneMsgRNA15E1

<400>  325
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa ccttctggcc ctagcagggc tggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacacccacg ccccagccct      600

ctgaaggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  326
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PMOEx1-TGEE5 ddPCR reference probe


<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  g is modified with ZEN

<400>  326
tctcagatgc atggtgga                                                     18


<210>  327
<211>  15
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PMOEx1-TGEE6 drop off probe


<220>
<221>  misc_feature
<222>  (8)..(8)
<223>  c is a locked nucleic acid

<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  c is a locked nucleic acid

<400>  327
ctcctggcca ttctg                                                        15


<210>  328
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PMOEx1 F2 FWD primer

<400>  328
ctgaggtaca aaggggga                                                     18


<210>  329
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PMOEx2 R1 REV primer

<400>  329
ggtctctgga acacaac                                                      17


<210>  330
<211>  1623
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, 2NLS variant, Construct 12863 and 13185

<400>  330

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Ile 
    1010                 1015                 1020             


Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln Ile Leu  Lys Glu Asn 
    1025                 1030                 1035             


Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu Lys Leu  Tyr Leu Tyr 
    1040                 1045                 1050             


Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val Asp Gln  Glu Leu Asp 
    1055                 1060                 1065             


Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp Ala Ile  Val Pro Gln 
    1070                 1075                 1080             


Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn Lys Val  Leu Thr Arg 
    1085                 1090                 1095             


Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn Val Pro  Ser Glu Glu 
    1100                 1105                 1110             


Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg Gln Leu  Leu Asn Ala 
    1115                 1120                 1125             


Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn Leu Thr  Lys Ala Glu 
    1130                 1135                 1140             


Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1145                 1150                 1155             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1160                 1165                 1170             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1175                 1180                 1185             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1190                 1195                 1200             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1205                 1210                 1215             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1220                 1225                 1230             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1235                 1240                 1245             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1250                 1255                 1260             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1265                 1270                 1275             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1280                 1285                 1290             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1295                 1300                 1305             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1310                 1315                 1320             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1325                 1330                 1335             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1340                 1345                 1350             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1355                 1360                 1365             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1370                 1375                 1380             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1385                 1390                 1395             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1400                 1405                 1410             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1415                 1420                 1425             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1430                 1435                 1440             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1445                 1450                 1455             


Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro Gln His  Leu Val Arg 
    1460                 1465                 1470             


Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn 
    1475                 1480                 1485             


Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile 
    1490                 1495                 1500             


Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn 
    1505                 1510                 1515             


Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala 
    1520                 1525                 1530             


Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu 
    1535                 1540                 1545             


Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly Phe Thr  Phe Leu Asp 
    1550                 1555                 1560             


Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr Gln Thr  Val Thr Glu 
    1565                 1570                 1575             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1580                 1585                 1590             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1595                 1600                 1605             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1610                 1615                 1620             


<210>  331
<211>  1504
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, 2NLS variant, Construct 13190

<400>  331

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1340                 1345                 1350             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1355                 1360                 1365             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1370                 1375                 1380             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1385                 1390                 1395             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1400                 1405                 1410             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1415                 1420                 1425             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu 
    1430                 1435                 1440             


Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr 
    1445                 1450                 1455             


Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu 
    1460                 1465                 1470             


Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg 
    1475                 1480                 1485             


Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala Lys Lys  Lys Lys Gly 
    1490                 1495                 1500             


Ala 
    


<210>  332
<211>  1398
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta, PAMBD whole delta, 2NLS variant, Construct 
       13191

<400>  332

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Gly Gly  Ser Gly Tyr Gln Thr  Val Thr Glu 
    1340                 1345                 1350             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1355                 1360                 1365             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1370                 1375                 1380             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1385                 1390                 1395             


<210>  333
<211>  1491
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta, PAMBD loop delta, 2NLS variant, Construct 
       13192

<400>  333

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1340                 1345                 1350             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1355                 1360                 1365             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1370                 1375                 1380             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1385                 1390                 1395             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1400                 1405                 1410             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1415                 1420                 1425             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly Ser Gly  Tyr Gln Thr 
    1430                 1435                 1440             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1445                 1450                 1455             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1460                 1465                 1470             


Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys 
    1475                 1480                 1485             


Lys Gly  Ala 
    1490     


<210>  334
<211>  1650
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, Zinc finger PAMBD loop replacement longer linkers, 
       2NLS variant, Construct 12869

<400>  334

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Ile 
    1010                 1015                 1020             


Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln Ile Leu  Lys Glu Asn 
    1025                 1030                 1035             


Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu Lys Leu  Tyr Leu Tyr 
    1040                 1045                 1050             


Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val Asp Gln  Glu Leu Asp 
    1055                 1060                 1065             


Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp Ala Ile  Val Pro Gln 
    1070                 1075                 1080             


Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn Lys Val  Leu Thr Arg 
    1085                 1090                 1095             


Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn Val Pro  Ser Glu Glu 
    1100                 1105                 1110             


Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg Gln Leu  Leu Asn Ala 
    1115                 1120                 1125             


Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn Leu Thr  Lys Ala Glu 
    1130                 1135                 1140             


Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1145                 1150                 1155             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1160                 1165                 1170             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1175                 1180                 1185             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1190                 1195                 1200             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1205                 1210                 1215             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1220                 1225                 1230             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1235                 1240                 1245             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1250                 1255                 1260             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1265                 1270                 1275             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1280                 1285                 1290             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1295                 1300                 1305             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1310                 1315                 1320             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1325                 1330                 1335             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1340                 1345                 1350             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1355                 1360                 1365             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1370                 1375                 1380             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1385                 1390                 1395             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1400                 1405                 1410             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1415                 1420                 1425             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1430                 1435                 1440             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1445                 1450                 1455             


Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro Gln His  Leu Val Arg 
    1460                 1465                 1470             


Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn 
    1475                 1480                 1485             


Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile 
    1490                 1495                 1500             


Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn 
    1505                 1510                 1515             


Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala 
    1520                 1525                 1530             


Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu 
    1535                 1540                 1545             


Lys Tyr  Thr Ser Phe Gly Ala  Gly Gly Ser Ala Gly  Gly Thr Gly 
    1550                 1555                 1560             


Gly Ser  Ala Gly Gly Ser Gly  Ala Pro Tyr Arg Cys  Pro Cys Arg 
    1565                 1570                 1575             


Ala Gly  Cys Pro Ser Leu Ala  Ser Met Gln Ala His  Met Arg Gly 
    1580                 1585                 1590             


His Ser  Gly Gly Ser Gly Tyr  Gln Thr Val Thr Glu  Val Leu Asp 
    1595                 1600                 1605             


Ala Thr  Leu Ile Tyr Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1610                 1615                 1620             


Thr Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1625                 1630                 1635             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Gly Ala  
    1640                 1645                 1650 


<210>  335
<211>  1567
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, Lac repressor DBD PAMBD whole replacement, 2NLS 
       variant, Construct 12870

<400>  335

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Ile 
    1010                 1015                 1020             


Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln Ile Leu  Lys Glu Asn 
    1025                 1030                 1035             


Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu Lys Leu  Tyr Leu Tyr 
    1040                 1045                 1050             


Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val Asp Gln  Glu Leu Asp 
    1055                 1060                 1065             


Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp Ala Ile  Val Pro Gln 
    1070                 1075                 1080             


Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn Lys Val  Leu Thr Arg 
    1085                 1090                 1095             


Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn Val Pro  Ser Glu Glu 
    1100                 1105                 1110             


Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg Gln Leu  Leu Asn Ala 
    1115                 1120                 1125             


Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn Leu Thr  Lys Ala Glu 
    1130                 1135                 1140             


Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1145                 1150                 1155             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1160                 1165                 1170             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1175                 1180                 1185             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1190                 1195                 1200             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1205                 1210                 1215             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1220                 1225                 1230             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1235                 1240                 1245             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1250                 1255                 1260             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1265                 1270                 1275             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1280                 1285                 1290             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1295                 1300                 1305             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1310                 1315                 1320             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1325                 1330                 1335             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1340                 1345                 1350             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1355                 1360                 1365             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1370                 1375                 1380             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1385                 1390                 1395             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1400                 1405                 1410             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1415                 1420                 1425             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1430                 1435                 1440             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1445                 1450                 1455             


Leu Gln  Lys Ala Gly Gly Ser  Gly Met Lys Pro Val  Thr Leu Tyr 
    1460                 1465                 1470             


Asp Val  Ala Glu Tyr Ala Gly  Val Ser Tyr Gln Thr  Val Ser Arg 
    1475                 1480                 1485             


Val Val  Asn Gln Ala Ser His  Val Ser Ala Lys Thr  Arg Glu Lys 
    1490                 1495                 1500             


Val Glu  Ala Ala Met Ala Glu  Leu Asn Gly Gly Ser  Gly Tyr Gln 
    1505                 1510                 1515             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1520                 1525                 1530             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1535                 1540                 1545             


Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys 
    1550                 1555                 1560             


Lys Lys  Gly Ala 
    1565         


<210>  336
<211>  1587
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SSO7D PAMBD whole replacement, 2NLS variant, Construct
       12878

<400>  336

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Ile 
    1010                 1015                 1020             


Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln Ile Leu  Lys Glu Asn 
    1025                 1030                 1035             


Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu Lys Leu  Tyr Leu Tyr 
    1040                 1045                 1050             


Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val Asp Gln  Glu Leu Asp 
    1055                 1060                 1065             


Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp Ala Ile  Val Pro Gln 
    1070                 1075                 1080             


Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn Lys Val  Leu Thr Arg 
    1085                 1090                 1095             


Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn Val Pro  Ser Glu Glu 
    1100                 1105                 1110             


Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg Gln Leu  Leu Asn Ala 
    1115                 1120                 1125             


Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn Leu Thr  Lys Ala Glu 
    1130                 1135                 1140             


Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1145                 1150                 1155             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1160                 1165                 1170             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1175                 1180                 1185             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1190                 1195                 1200             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1205                 1210                 1215             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1220                 1225                 1230             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1235                 1240                 1245             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1250                 1255                 1260             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1265                 1270                 1275             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1280                 1285                 1290             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1295                 1300                 1305             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1310                 1315                 1320             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1325                 1330                 1335             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1340                 1345                 1350             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1355                 1360                 1365             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1370                 1375                 1380             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1385                 1390                 1395             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1400                 1405                 1410             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1415                 1420                 1425             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1430                 1435                 1440             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1445                 1450                 1455             


Leu Gln  Lys Ala Gly Gly Ser  Ala Gly Met Ala Thr  Val Lys Phe 
    1460                 1465                 1470             


Lys Tyr  Lys Gly Glu Glu Lys  Glu Val Asp Ile Ser  Lys Ile Lys 
    1475                 1480                 1485             


Lys Val  Trp Arg Val Gly Lys  Met Ile Ser Phe Thr  Tyr Asp Glu 
    1490                 1495                 1500             


Gly Gly  Gly Lys Thr Gly Arg  Gly Ala Val Ser Glu  Lys Asp Ala 
    1505                 1510                 1515             


Pro Lys  Glu Leu Leu Gln Met  Leu Glu Lys Gln Lys  Lys Gly Gly 
    1520                 1525                 1530             


Ser Ala  Gly Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1535                 1540                 1545             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1550                 1555                 1560             


Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala 
    1565                 1570                 1575             


Gly Gln  Ala Lys Lys Lys Lys  Gly Ala 
    1580                 1585         


<210>  337
<211>  1597
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SSO7D PAMBD whole replacement longer linkers, 2NLS 
       variant, Construct 12880

<400>  337

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Ile 
    1010                 1015                 1020             


Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln Ile Leu  Lys Glu Asn 
    1025                 1030                 1035             


Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu Lys Leu  Tyr Leu Tyr 
    1040                 1045                 1050             


Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val Asp Gln  Glu Leu Asp 
    1055                 1060                 1065             


Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp Ala Ile  Val Pro Gln 
    1070                 1075                 1080             


Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn Lys Val  Leu Thr Arg 
    1085                 1090                 1095             


Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn Val Pro  Ser Glu Glu 
    1100                 1105                 1110             


Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg Gln Leu  Leu Asn Ala 
    1115                 1120                 1125             


Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn Leu Thr  Lys Ala Glu 
    1130                 1135                 1140             


Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1145                 1150                 1155             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1160                 1165                 1170             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1175                 1180                 1185             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1190                 1195                 1200             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1205                 1210                 1215             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1220                 1225                 1230             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1235                 1240                 1245             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1250                 1255                 1260             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1265                 1270                 1275             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1280                 1285                 1290             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1295                 1300                 1305             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1310                 1315                 1320             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1325                 1330                 1335             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1340                 1345                 1350             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1355                 1360                 1365             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1370                 1375                 1380             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1385                 1390                 1395             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1400                 1405                 1410             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1415                 1420                 1425             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1430                 1435                 1440             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1445                 1450                 1455             


Leu Gln  Lys Ala Gly Gly Ser  Ala Gly Gly Thr Gly  Gly Ser Met 
    1460                 1465                 1470             


Ala Thr  Val Lys Phe Lys Tyr  Lys Gly Glu Glu Lys  Glu Val Asp 
    1475                 1480                 1485             


Ile Ser  Lys Ile Lys Lys Val  Trp Arg Val Gly Lys  Met Ile Ser 
    1490                 1495                 1500             


Phe Thr  Tyr Asp Glu Gly Gly  Gly Lys Thr Gly Arg  Gly Ala Val 
    1505                 1510                 1515             


Ser Glu  Lys Asp Ala Pro Lys  Glu Leu Leu Gln Met  Leu Glu Lys 
    1520                 1525                 1530             


Gln Lys  Lys Gly Gly Ser Ala  Gly Gly Thr Gly Gly  Ser Tyr Gln 
    1535                 1540                 1545             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1550                 1555                 1560             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1565                 1570                 1575             


Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys 
    1580                 1585                 1590             


Lys Lys  Gly Ala 
    1595         


<210>  338
<211>  1477
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta, SSO7D PAMBD whole replacement longer 
       linkers, 2NLS variant, Construct 13207

<400>  338

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser 
    1340                 1345                 1350             


Met Ala  Thr Val Lys Phe Lys  Tyr Lys Gly Glu Glu  Lys Glu Val 
    1355                 1360                 1365             


Asp Ile  Ser Lys Ile Lys Lys  Val Trp Arg Val Gly  Lys Met Ile 
    1370                 1375                 1380             


Ser Phe  Thr Tyr Asp Glu Gly  Gly Gly Lys Thr Gly  Arg Gly Ala 
    1385                 1390                 1395             


Val Ser  Glu Lys Asp Ala Pro  Lys Glu Leu Leu Gln  Met Leu Glu 
    1400                 1405                 1410             


Lys Gln  Lys Lys Gly Gly Ser  Ala Gly Gly Thr Gly  Gly Ser Tyr 
    1415                 1420                 1425             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1430                 1435                 1440             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1445                 1450                 1455             


Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys 
    1460                 1465                 1470             


Lys Lys  Lys Gly 
    1475         


<210>  339
<211>  1690
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, STO7D PAMBD loop replacement longer linker, 2NLS 
       variant, Construct 12885

<400>  339

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Ile 
    1010                 1015                 1020             


Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln Ile Leu  Lys Glu Asn 
    1025                 1030                 1035             


Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu Lys Leu  Tyr Leu Tyr 
    1040                 1045                 1050             


Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val Asp Gln  Glu Leu Asp 
    1055                 1060                 1065             


Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp Ala Ile  Val Pro Gln 
    1070                 1075                 1080             


Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn Lys Val  Leu Thr Arg 
    1085                 1090                 1095             


Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn Val Pro  Ser Glu Glu 
    1100                 1105                 1110             


Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg Gln Leu  Leu Asn Ala 
    1115                 1120                 1125             


Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn Leu Thr  Lys Ala Glu 
    1130                 1135                 1140             


Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1145                 1150                 1155             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1160                 1165                 1170             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1175                 1180                 1185             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1190                 1195                 1200             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1205                 1210                 1215             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1220                 1225                 1230             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1235                 1240                 1245             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1250                 1255                 1260             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1265                 1270                 1275             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1280                 1285                 1290             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1295                 1300                 1305             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1310                 1315                 1320             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1325                 1330                 1335             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1340                 1345                 1350             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1355                 1360                 1365             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1370                 1375                 1380             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1385                 1390                 1395             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1400                 1405                 1410             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1415                 1420                 1425             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1430                 1435                 1440             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1445                 1450                 1455             


Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro Gln His  Leu Val Arg 
    1460                 1465                 1470             


Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn 
    1475                 1480                 1485             


Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile 
    1490                 1495                 1500             


Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn 
    1505                 1510                 1515             


Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala 
    1520                 1525                 1530             


Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu 
    1535                 1540                 1545             


Lys Tyr  Thr Ser Phe Gly Ala  Gly Gly Ser Ala Gly  Gly Thr Gly 
    1550                 1555                 1560             


Gly Ser  Met Val Thr Val Lys  Phe Lys Tyr Lys Gly  Glu Glu Lys 
    1565                 1570                 1575             


Glu Val  Asp Ile Ser Lys Ile  Lys Lys Val Trp Arg  Val Gly Lys 
    1580                 1585                 1590             


Met Ile  Ser Phe Thr Tyr Asp  Asp Asn Gly Lys Thr  Gly Arg Gly 
    1595                 1600                 1605             


Ala Val  Ser Glu Lys Asp Ala  Pro Lys Glu Leu Leu  Gln Met Leu 
    1610                 1615                 1620             


Glu Lys  Ser Gly Lys Lys Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1625                 1630                 1635             


Ser Tyr  Gln Thr Val Thr Glu  Val Leu Asp Ala Thr  Leu Ile Tyr 
    1640                 1645                 1650             


Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg Thr Asp  Leu Ser Gln 
    1655                 1660                 1665             


Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr Lys Lys  Ala Gly Gln 
    1670                 1675                 1680             


Ala Lys  Lys Lys Lys Gly Ala  
    1685                 1690 


<210>  340
<211>  1463
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta, HIV integrase DBD PAMBD replacement whole 
       replacement longer linker, 2NLS variant, Construct 13203

<400>  340

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser 
    1340                 1345                 1350             


Ile Gln  Asn Phe Arg Val Tyr  Tyr Arg Asp Ser Arg  Asn Pro Leu 
    1355                 1360                 1365             


Trp Lys  Gly Pro Ala Lys Leu  Leu Trp Lys Gly Glu  Gly Ala Trp 
    1370                 1375                 1380             


Ile Gln  Asp Asn Ser Asp Ile  Lys Trp Pro Arg Arg  Lys Ala Lys 
    1385                 1390                 1395             


Ile Ile  Arg Asp Gly Gly Ser  Ala Gly Gly Thr Gly  Gly Ser Tyr 
    1400                 1405                 1410             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1415                 1420                 1425             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1430                 1435                 1440             


Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys 
    1445                 1450                 1455             


Lys Lys  Lys Gly Ala 
    1460             


<210>  341
<211>  1556
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta, HIV integrase DBD PAMBD replacement loop 
       replacement longer linker, 2NLS variant, Construct 13204

<400>  341

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1340                 1345                 1350             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1355                 1360                 1365             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1370                 1375                 1380             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1385                 1390                 1395             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1400                 1405                 1410             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1415                 1420                 1425             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly Ser Ala  Gly Gly Thr 
    1430                 1435                 1440             


Gly Gly  Ser Ile Gln Asn Phe  Arg Val Tyr Tyr Arg  Asp Ser Arg 
    1445                 1450                 1455             


Asn Pro  Leu Trp Lys Gly Pro  Ala Lys Leu Leu Trp  Lys Gly Glu 
    1460                 1465                 1470             


Gly Ala  Trp Ile Gln Asp Asn  Ser Asp Ile Lys Trp  Pro Arg Arg 
    1475                 1480                 1485             


Lys Ala  Lys Ile Ile Arg Asp  Gly Gly Ser Ala Gly  Gly Thr Gly 
    1490                 1495                 1500             


Gly Ser  Tyr Gln Thr Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile 
    1505                 1510                 1515             


Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser 
    1520                 1525                 1530             


Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly 
    1535                 1540                 1545             


Gln Ala  Lys Lys Lys Lys Gly  Ala 
    1550                 1555     


<210>  342
<211>  1507
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with longer linkers, 2NLS, Construct 14001

<400>  342

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1490                 1495                 1500             


Lys Lys  Lys Lys 
    1505         


<210>  343
<211>  1610
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta replaced with SSB, 2NLS, Construct 14002

<400>  343

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Thr Thr  Glu Thr Phe Val Lys  Asp Ile Lys Pro Gly  Leu Lys Asn 
    1025                 1030                 1035             


Leu Asn  Leu Ile Phe Ile Val  Leu Glu Thr Gly Arg  Val Thr Lys 
    1040                 1045                 1050             


Thr Lys  Asp Gly His Glu Val  Arg Thr Cys Lys Val  Ala Asp Lys 
    1055                 1060                 1065             


Thr Gly  Ser Ile Asn Ile Ser  Val Trp Asp Asp Val  Gly Asn Leu 
    1070                 1075                 1080             


Ile Gln  Pro Gly Asp Ile Ile  Arg Leu Thr Lys Gly  Tyr Ala Ser 
    1085                 1090                 1095             


Val Phe  Lys Gly Cys Leu Thr  Leu Tyr Thr Gly Arg  Gly Gly Asp 
    1100                 1105                 1110             


Leu Gln  Lys Ile Gly Glu Phe  Cys Met Val Tyr Ser  Glu Val Pro 
    1115                 1120                 1125             


Asn Phe  Ser Glu Pro Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Ala Thr  Val Arg Lys Val Leu  Ala Met Pro Gln Val  Asn Ile Val 
    1310                 1315                 1320             


Lys Lys  Thr Glu Val Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile 
    1325                 1330                 1335             


Leu Ser  Lys Arg Glu Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly 
    1340                 1345                 1350             


Trp Asp  Thr Arg Lys Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala 
    1355                 1360                 1365             


Tyr Ser  Ile Leu Val Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys 
    1370                 1375                 1380             


Lys Leu  Lys Ser Val Lys Val  Leu Val Gly Ile Thr  Ile Met Glu 
    1385                 1390                 1395             


Lys Gly  Ser Tyr Glu Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys 
    1400                 1405                 1410             


Gly Tyr  Lys Asp Ile Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys 
    1415                 1420                 1425             


Tyr Ser  Leu Phe Glu Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala 
    1430                 1435                 1440             


Ser Ala  Thr Glu Leu Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln 
    1445                 1450                 1455             


His Leu  Val Arg Leu Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr 
    1460                 1465                 1470             


Thr Gly  Ser Asn Asn Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu 
    1475                 1480                 1485             


Phe Lys  Glu Ile Phe Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr 
    1490                 1495                 1500             


Ile Leu  Lys Asn Lys Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp 
    1505                 1510                 1515             


Glu Gln  Phe Ala Val Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe 
    1520                 1525                 1530             


Val Ser  Leu Leu Lys Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe 
    1535                 1540                 1545             


Thr Phe  Leu Asp Leu Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln 
    1550                 1555                 1560             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1565                 1570                 1575             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1580                 1585                 1590             


Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys 
    1595                 1600                 1605             


Lys Lys  
    1610 


<210>  344
<211>  1482
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with larger deletion, 2NLS, Construct 14003

<400>  344

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Gly Gly Ser  Gly Gly Leu 
        995                 1000                 1005             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1010                 1015                 1020             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1025                 1030                 1035             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1040                 1045                 1050             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1055                 1060                 1065             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1070                 1075                 1080             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1085                 1090                 1095             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1100                 1105                 1110             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1115                 1120                 1125             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1130                 1135                 1140             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1145                 1150                 1155             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1160                 1165                 1170             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1175                 1180                 1185             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1190                 1195                 1200             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1205                 1210                 1215             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1220                 1225                 1230             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1235                 1240                 1245             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1250                 1255                 1260             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1265                 1270                 1275             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1280                 1285                 1290             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1295                 1300                 1305             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1310                 1315                 1320             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1325                 1330                 1335             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1340                 1345                 1350             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1355                 1360                 1365             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1370                 1375                 1380             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1385                 1390                 1395             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1400                 1405                 1410             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1415                 1420                 1425             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1430                 1435                 1440             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1445                 1450                 1455             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1460                 1465                 1470             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1475                 1480         


<210>  345
<211>  1610
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-LoopDel, 2NLS variant

<400>  345

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile 
225                 230                 235                 240 


Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val 
                245                 250                 255     


Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile 
            260                 265                 270         


Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala 
        275                 280                 285             


Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg 
    290                 295                 300                 


Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala 
305                 310                 315                 320 


Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val 
                325                 330                 335     


Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala 
            340                 345                 350         


Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg 
        355                 360                 365             


Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr 
    370                 375                 380                 


Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu 
385                 390                 395                 400 


Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln 
                405                 410                 415     


Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu 
            420                 425                 430         


Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser 
        435                 440                 445             


Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn 
    450                 455                 460                 


Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn 
465                 470                 475                 480 


Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser 
                485                 490                 495     


Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly 
            500                 505                 510         


Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala 
        515                 520                 525             


Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala 
    530                 535                 540                 


Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp 
545                 550                 555                 560 


Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr 
                565                 570                 575     


Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val 
            580                 585                 590         


Gly Ile Gly Ala Thr Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile 
        595                 600                 605             


Leu Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg 
    610                 615                 620                 


Asp Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro 
625                 630                 635                 640 


His Gln Ile His Leu Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu 
                645                 650                 655     


Glu Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile 
            660                 665                 670         


Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn 
        675                 680                 685             


Ser Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro 
    690                 695                 700                 


Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe 
705                 710                 715                 720 


Ile Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val 
                725                 730                 735     


Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu 
            740                 745                 750         


Leu Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe 
        755                 760                 765             


Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr 
    770                 775                 780                 


Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys 
785                 790                 795                 800 


Ile Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe 
                805                 810                 815     


Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp 
            820                 825                 830         


Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile 
        835                 840                 845             


Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg 
    850                 855                 860                 


Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu 
865                 870                 875                 880 


Lys Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile 
                885                 890                 895     


Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu 
            900                 905                 910         


Lys Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp 
        915                 920                 925             


Asp Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly 
    930                 935                 940                 


Gln Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro 
945                 950                 955                 960 


Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu 
                965                 970                 975     


Val Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
            980                 985                 990         


Arg Glu Asn Gln Thr Thr Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg 
        995                 1000                 1005             


Lys Lys  Arg Ile Glu Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile 
    1010                 1015                 1020             


Leu Lys  Glu Asn Pro Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys 
    1025                 1030                 1035             


Leu Tyr  Leu Tyr Tyr Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp 
    1040                 1045                 1050             


Gln Glu  Leu Asp Ile Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala 
    1055                 1060                 1065             


Ile Val  Pro Gln Ser Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys 
    1070                 1075                 1080             


Val Leu  Thr Arg Ser Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val 
    1085                 1090                 1095             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1100                 1105                 1110             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1115                 1120                 1125             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Ala Thr  Val Arg Lys Val Leu  Ala Met Pro Gln Val  Asn Ile Val 
    1310                 1315                 1320             


Lys Lys  Thr Glu Val Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile 
    1325                 1330                 1335             


Leu Ser  Lys Arg Glu Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly 
    1340                 1345                 1350             


Trp Asp  Thr Arg Lys Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala 
    1355                 1360                 1365             


Tyr Ser  Ile Leu Val Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys 
    1370                 1375                 1380             


Lys Leu  Lys Ser Val Lys Val  Leu Val Gly Ile Thr  Ile Met Glu 
    1385                 1390                 1395             


Lys Gly  Ser Tyr Glu Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys 
    1400                 1405                 1410             


Gly Tyr  Lys Asp Ile Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys 
    1415                 1420                 1425             


Tyr Ser  Leu Phe Glu Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala 
    1430                 1435                 1440             


Ser Ala  Thr Glu Leu Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln 
    1445                 1450                 1455             


His Leu  Val Arg Leu Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr 
    1460                 1465                 1470             


Thr Gly  Ser Asn Asn Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu 
    1475                 1480                 1485             


Phe Lys  Glu Ile Phe Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr 
    1490                 1495                 1500             


Ile Leu  Lys Asn Lys Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp 
    1505                 1510                 1515             


Glu Gln  Phe Ala Val Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe 
    1520                 1525                 1530             


Val Ser  Leu Leu Lys Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe 
    1535                 1540                 1545             


Thr Phe  Leu Asp Leu Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln 
    1550                 1555                 1560             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1565                 1570                 1575             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1580                 1585                 1590             


Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys 
    1595                 1600                 1605             


Lys Lys  
    1610 


<210>  346
<211>  1620
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-LoopQQ, 2NLS variant

<400>  346

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile 
225                 230                 235                 240 


Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val 
                245                 250                 255     


Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile 
            260                 265                 270         


Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala 
        275                 280                 285             


Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg 
    290                 295                 300                 


Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala 
305                 310                 315                 320 


Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val 
                325                 330                 335     


Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala 
            340                 345                 350         


Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg 
        355                 360                 365             


Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr 
    370                 375                 380                 


Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu 
385                 390                 395                 400 


Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln 
                405                 410                 415     


Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu 
            420                 425                 430         


Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser 
        435                 440                 445             


Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn 
    450                 455                 460                 


Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn 
465                 470                 475                 480 


Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser 
                485                 490                 495     


Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly 
            500                 505                 510         


Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala 
        515                 520                 525             


Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala 
    530                 535                 540                 


Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp 
545                 550                 555                 560 


Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr 
                565                 570                 575     


Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val 
            580                 585                 590         


Gly Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Ala Thr Gln 
        595                 600                 605             


Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
    610                 615                 620                 


Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys 
625                 630                 635                 640 


Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys 
                645                 650                 655     


Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu 
            660                 665                 670         


Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
        675                 680                 685             


Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu 
    690                 695                 700                 


Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val 
705                 710                 715                 720 


Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
                725                 730                 735     


Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu 
            740                 745                 750         


Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
        755                 760                 765             


Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys 
    770                 775                 780                 


Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
785                 790                 795                 800 


Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
                805                 810                 815     


Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
            820                 825                 830         


Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
        835                 840                 845             


Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
    850                 855                 860                 


Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
865                 870                 875                 880 


Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr 
                885                 890                 895     


Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys 
            900                 905                 910         


Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser 
        915                 920                 925             


Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys 
    930                 935                 940                 


Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His 
945                 950                 955                 960 


Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile 
                965                 970                 975     


Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His 
            980                 985                 990         


Lys Pro Glu Asn Ile Val Ile Glu  Met Ala Arg Glu Asn  Gln Thr Thr 
        995                 1000                 1005             


Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg Lys Lys  Arg Ile Glu 
    1010                 1015                 1020             


Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile Leu Lys  Glu Asn Pro 
    1025                 1030                 1035             


Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys Leu Tyr  Leu Tyr Tyr 
    1040                 1045                 1050             


Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp Gln Glu  Leu Asp Ile 
    1055                 1060                 1065             


Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala Ile Val  Pro Gln Ser 
    1070                 1075                 1080             


Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys Val Leu  Thr Arg Ser 
    1085                 1090                 1095             


Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val Pro Ser  Glu Glu Val 
    1100                 1105                 1110             


Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln Leu Leu  Asn Ala Lys 
    1115                 1120                 1125             


Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu Thr Lys  Ala Glu Arg 
    1130                 1135                 1140             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1145                 1150                 1155             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1160                 1165                 1170             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1175                 1180                 1185             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1190                 1195                 1200             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1205                 1210                 1215             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1220                 1225                 1230             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1235                 1240                 1245             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1250                 1255                 1260             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1265                 1270                 1275             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1280                 1285                 1290             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1295                 1300                 1305             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1310                 1315                 1320             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1325                 1330                 1335             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1340                 1345                 1350             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1355                 1360                 1365             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1370                 1375                 1380             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1385                 1390                 1395             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1400                 1405                 1410             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1415                 1420                 1425             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1430                 1435                 1440             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1445                 1450                 1455             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1460                 1465                 1470             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1475                 1480                 1485             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1490                 1495                 1500             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1505                 1510                 1515             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1520                 1525                 1530             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1535                 1540                 1545             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1550                 1555                 1560             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1565                 1570                 1575             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1580                 1585                 1590             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala 
    1595                 1600                 1605             


Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys Lys Lys  
    1610                 1615                 1620 


<210>  347
<211>  1620
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI-LoopAA, 2NLS variant

<400>  347

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile 
225                 230                 235                 240 


Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val 
                245                 250                 255     


Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile 
            260                 265                 270         


Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala 
        275                 280                 285             


Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg 
    290                 295                 300                 


Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala 
305                 310                 315                 320 


Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val 
                325                 330                 335     


Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala 
            340                 345                 350         


Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg 
        355                 360                 365             


Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr 
    370                 375                 380                 


Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu 
385                 390                 395                 400 


Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln 
                405                 410                 415     


Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu 
            420                 425                 430         


Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser 
        435                 440                 445             


Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn 
    450                 455                 460                 


Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn 
465                 470                 475                 480 


Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser 
                485                 490                 495     


Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly 
            500                 505                 510         


Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala 
        515                 520                 525             


Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala 
    530                 535                 540                 


Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp 
545                 550                 555                 560 


Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr 
                565                 570                 575     


Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val 
            580                 585                 590         


Gly Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Ala Thr Gln 
        595                 600                 605             


Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
    610                 615                 620                 


Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys 
625                 630                 635                 640 


Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys 
                645                 650                 655     


Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu 
            660                 665                 670         


Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
        675                 680                 685             


Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu 
    690                 695                 700                 


Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val 
705                 710                 715                 720 


Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
                725                 730                 735     


Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu 
            740                 745                 750         


Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
        755                 760                 765             


Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys 
    770                 775                 780                 


Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
785                 790                 795                 800 


Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
                805                 810                 815     


Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
            820                 825                 830         


Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
        835                 840                 845             


Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
    850                 855                 860                 


Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
865                 870                 875                 880 


Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr 
                885                 890                 895     


Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys 
            900                 905                 910         


Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser 
        915                 920                 925             


Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys 
    930                 935                 940                 


Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His 
945                 950                 955                 960 


Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile 
                965                 970                 975     


Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His 
            980                 985                 990         


Lys Pro Glu Asn Ile Val Ile Glu  Met Ala Arg Glu Asn  Gln Thr Thr 
        995                 1000                 1005             


Thr Lys  Gly Leu Gln Gln Ser  Arg Glu Arg Lys Lys  Arg Ile Glu 
    1010                 1015                 1020             


Glu Gly  Ile Lys Glu Leu Glu  Ser Gln Ile Leu Lys  Glu Asn Pro 
    1025                 1030                 1035             


Val Glu  Asn Thr Gln Leu Gln  Asn Glu Lys Leu Tyr  Leu Tyr Tyr 
    1040                 1045                 1050             


Leu Gln  Asn Gly Arg Asp Met  Tyr Val Asp Gln Glu  Leu Asp Ile 
    1055                 1060                 1065             


Asn Arg  Leu Ser Asp Tyr Asp  Val Asp Ala Ile Val  Pro Gln Ser 
    1070                 1075                 1080             


Phe Ile  Lys Asp Asp Ser Ile  Asp Asn Lys Val Leu  Thr Arg Ser 
    1085                 1090                 1095             


Val Glu  Asn Arg Gly Lys Ser  Asp Asn Val Pro Ser  Glu Glu Val 
    1100                 1105                 1110             


Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln Leu Leu  Asn Ala Lys 
    1115                 1120                 1125             


Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu Thr Lys  Ala Glu Arg 
    1130                 1135                 1140             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1145                 1150                 1155             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1160                 1165                 1170             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1175                 1180                 1185             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1190                 1195                 1200             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1205                 1210                 1215             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1220                 1225                 1230             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1235                 1240                 1245             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1250                 1255                 1260             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1265                 1270                 1275             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1280                 1285                 1290             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1295                 1300                 1305             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1310                 1315                 1320             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1325                 1330                 1335             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1340                 1345                 1350             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1355                 1360                 1365             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1370                 1375                 1380             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1385                 1390                 1395             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1400                 1405                 1410             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1415                 1420                 1425             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1430                 1435                 1440             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1445                 1450                 1455             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1460                 1465                 1470             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1475                 1480                 1485             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1490                 1495                 1500             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1505                 1510                 1515             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1520                 1525                 1530             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1535                 1540                 1545             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1550                 1555                 1560             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1565                 1570                 1575             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1580                 1585                 1590             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala 
    1595                 1600                 1605             


Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys Lys Lys  
    1610                 1615                 1620 


<210>  348
<211>  1609
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ScLoop deltaSp, SV40+nucleoplasminNLS (TG14481)

<400>  348

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Ile Glu Glu Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu 
    1010                 1015                 1020             


Lys Glu  Asn Pro Val Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu 
    1025                 1030                 1035             


Tyr Leu  Tyr Tyr Leu Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln 
    1040                 1045                 1050             


Glu Leu  Asp Ile Asn Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile 
    1055                 1060                 1065             


Val Pro  Gln Ser Phe Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val 
    1070                 1075                 1080             


Leu Thr  Arg Ser Val Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro 
    1085                 1090                 1095             


Ser Glu  Glu Val Val Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu 
    1100                 1105                 1110             


Leu Asn  Ala Lys Leu Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr 
    1115                 1120                 1125             


Lys Ala  Glu Arg Gly Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe 
    1130                 1135                 1140             


Ile Lys  Arg Gln Leu Val Glu  Thr Arg Gln Ile Thr  Lys His Val 
    1145                 1150                 1155             


Ala Arg  Ile Leu Asp Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn 
    1160                 1165                 1170             


Asp Lys  Pro Ile Arg Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys 
    1175                 1180                 1185             


Leu Val  Ser Asp Phe Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg 
    1190                 1195                 1200             


Asp Ile  Asn Asn Tyr His His  Ala His Asp Ala Tyr  Leu Asn Ala 
    1205                 1210                 1215             


Val Val  Gly Thr Ala Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser 
    1220                 1225                 1230             


Glu Phe  Val Tyr Gly Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met 
    1235                 1240                 1245             


Ile Ala  Lys Ser Glu Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg 
    1250                 1255                 1260             


Phe Phe  Tyr Ser Asn Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys 
    1265                 1270                 1275             


Leu Ala  Asn Gly Glu Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn 
    1280                 1285                 1290             


Gly Glu  Thr Gly Glu Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala 
    1295                 1300                 1305             


Thr Val  Arg Lys Val Leu Ala  Met Pro Gln Val Asn  Ile Val Lys 
    1310                 1315                 1320             


Lys Thr  Glu Val Gln Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu 
    1325                 1330                 1335             


Ser Lys  Arg Glu Ser Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp 
    1340                 1345                 1350             


Asp Thr  Arg Lys Tyr Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr 
    1355                 1360                 1365             


Ser Ile  Leu Val Val Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys 
    1370                 1375                 1380             


Leu Lys  Ser Val Lys Val Leu  Val Gly Ile Thr Ile  Met Glu Lys 
    1385                 1390                 1395             


Gly Ser  Tyr Glu Lys Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly 
    1400                 1405                 1410             


Tyr Lys  Asp Ile Lys Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr 
    1415                 1420                 1425             


Ser Leu  Phe Glu Leu Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser 
    1430                 1435                 1440             


Ala Thr  Glu Leu Gln Lys Ala  Asn Glu Leu Val Leu  Pro Gln His 
    1445                 1450                 1455             


Leu Val  Arg Leu Leu Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr 
    1460                 1465                 1470             


Gly Ser  Asn Asn Leu Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe 
    1475                 1480                 1485             


Lys Glu  Ile Phe Glu Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile 
    1490                 1495                 1500             


Leu Lys  Asn Lys Val Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu 
    1505                 1510                 1515             


Gln Phe  Ala Val Ser Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val 
    1520                 1525                 1530             


Ser Leu  Leu Lys Tyr Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr 
    1535                 1540                 1545             


Phe Leu  Asp Leu Asp Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr 
    1550                 1555                 1560             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1565                 1570                 1575             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1580                 1585                 1590             


Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys 
    1595                 1600                 1605             


Lys 
    


<210>  349
<211>  1507
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with positively charged linker, 2NLS, 
       Construct 14004

<400>  349

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Lys 
    1010                 1015                 1020             


Ala Arg  Gly Lys Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1490                 1495                 1500             


Lys Lys  Lys Lys 
    1505         


<210>  350
<211>  1500
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with short positively charged linker, 2NLS, 
       Construct 14005

<400>  350

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Lys Ser 
    1010                 1015                 1020             


Lys Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1430                 1435                 1440             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1445                 1450                 1455             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1460                 1465                 1470             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala 
    1475                 1480                 1485             


Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys Lys Lys  
    1490                 1495                 1500 


<210>  351
<211>  1493
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with H-NS linker, 2NLS, Construct 14007

<400>  351

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Ala Ala Val  Lys Ser Gly 
        995                 1000                 1005             


Thr Lys  Ala Lys Arg Ala Gln  Arg Pro Gly Leu Ser  Glu Ala Asp 
    1010                 1015                 1020             


Lys Ala  Gly Phe Ile Lys Arg  Gln Leu Val Glu Thr  Arg Gln Ile 
    1025                 1030                 1035             


Thr Lys  His Val Ala Arg Ile  Leu Asp Ser Arg Met  Asn Thr Lys 
    1040                 1045                 1050             


Arg Asp  Lys Asn Asp Lys Pro  Ile Arg Glu Val Lys  Val Ile Thr 
    1055                 1060                 1065             


Leu Lys  Ser Lys Leu Val Ser  Asp Phe Arg Lys Asp  Phe Gln Leu 
    1070                 1075                 1080             


Tyr Lys  Val Arg Asp Ile Asn  Asn Tyr His His Ala  His Asp Ala 
    1085                 1090                 1095             


Tyr Leu  Asn Ala Val Val Gly  Thr Ala Leu Ile Lys  Lys Tyr Pro 
    1100                 1105                 1110             


Lys Leu  Glu Ser Glu Phe Val  Tyr Gly Asp Tyr Lys  Val Tyr Asp 
    1115                 1120                 1125             


Val Arg  Lys Met Ile Ala Lys  Ser Glu Gln Glu Ile  Gly Lys Ala 
    1130                 1135                 1140             


Thr Ala  Lys Arg Phe Phe Tyr  Ser Asn Ile Met Asn  Phe Phe Lys 
    1145                 1150                 1155             


Thr Glu  Val Lys Leu Ala Asn  Gly Glu Ile Arg Lys  Arg Pro Leu 
    1160                 1165                 1170             


Ile Glu  Thr Asn Gly Glu Thr  Gly Glu Val Val Trp  Asn Lys Glu 
    1175                 1180                 1185             


Lys Asp  Phe Ala Thr Val Arg  Lys Val Leu Ala Met  Pro Gln Val 
    1190                 1195                 1200             


Asn Ile  Val Lys Lys Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys 
    1205                 1210                 1215             


Glu Ser  Ile Leu Ser Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg 
    1220                 1225                 1230             


Lys Lys  Gly Trp Asp Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro 
    1235                 1240                 1245             


Thr Val  Ala Tyr Ser Ile Leu  Val Val Ala Lys Val  Glu Lys Gly 
    1250                 1255                 1260             


Lys Ala  Lys Lys Leu Lys Ser  Val Lys Val Leu Val  Gly Ile Thr 
    1265                 1270                 1275             


Ile Met  Glu Lys Gly Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu 
    1280                 1285                 1290             


Glu Ala  Lys Gly Tyr Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys 
    1295                 1300                 1305             


Leu Pro  Lys Tyr Ser Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg 
    1310                 1315                 1320             


Met Leu  Ala Ser Ala Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val 
    1325                 1330                 1335             


Leu Pro  Gln His Leu Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile 
    1340                 1345                 1350             


Ser Ala  Thr Thr Gly Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His 
    1355                 1360                 1365             


Arg Glu  Glu Phe Lys Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser 
    1370                 1375                 1380             


Glu Lys  Tyr Ile Leu Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser 
    1385                 1390                 1395             


Ser Phe  Asp Glu Gln Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser 
    1400                 1405                 1410             


Asn Ser  Phe Val Ser Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Ser 
    1415                 1420                 1425             


Gly Gly  Phe Thr Phe Leu Asp  Leu Asp Val Lys Gln  Gly Arg Leu 
    1430                 1435                 1440             


Arg Tyr  Gln Thr Val Thr Glu  Val Leu Asp Ala Thr  Leu Ile Tyr 
    1445                 1450                 1455             


Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg Thr Asp  Leu Ser Gln 
    1460                 1465                 1470             


Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr Lys Lys  Ala Gly Gln 
    1475                 1480                 1485             


Ala Lys  Lys Lys Lys 
    1490             


<210>  352
<211>  871
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok Intein, C-terminal, Construct 14806

<400>  352

Met Ala Val Thr Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro 
1               5                   10                  15      


Lys Lys Lys Arg Lys Val Ile Lys Ile Ala Thr Arg Lys Tyr Leu Gly 
            20                  25                  30          


Lys Gln Asn Val Tyr Asp Ile Gly Val Glu Arg Asp His Asn Phe Ala 
        35                  40                  45              


Leu Lys Asn Gly Phe Ile Ala Ser Asn Cys Phe Asp Ser Val Glu Ile 
    50                  55                  60                  


Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
65                  70                  75                  80  


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
                85                  90                  95      


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
            100                 105                 110         


Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp 
        115                 120                 125             


Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly Trp Gly 
    130                 135                 140                 


Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly 
145                 150                 155                 160 


Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn Arg Asn 
                165                 170                 175     


Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Glu Ile 
            180                 185                 190         


Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu Gln Ile 
        195                 200                 205             


Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr 
    210                 215                 220                 


Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys Pro Glu 
225                 230                 235                 240 


Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Thr Lys Gly 
                245                 250                 255     


Leu Gln Gln Ser Arg Glu Arg Lys Lys Arg Ile Glu Glu Gly Ile Lys 
            260                 265                 270         


Glu Leu Glu Ser Gln Ile Leu Lys Glu Asn Pro Val Glu Asn Thr Gln 
        275                 280                 285             


Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp 
    290                 295                 300                 


Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp 
305                 310                 315                 320 


Val Asp Ala Ile Val Pro Gln Ser Phe Ile Lys Asp Asp Ser Ile Asp 
                325                 330                 335     


Asn Lys Val Leu Thr Arg Ser Val Glu Asn Arg Gly Lys Ser Asp Asn 
            340                 345                 350         


Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln 
        355                 360                 365             


Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr 
    370                 375                 380                 


Lys Ala Glu Arg Gly Gly Leu Ser Glu Ala Asp Lys Ala Gly Phe Ile 
385                 390                 395                 400 


Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Arg 
                405                 410                 415     


Ile Leu Asp Ser Arg Met Asn Thr Lys Arg Asp Lys Asn Asp Lys Pro 
            420                 425                 430         


Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp 
        435                 440                 445             


Phe Arg Lys Asp Phe Gln Leu Tyr Lys Val Arg Asp Ile Asn Asn Tyr 
    450                 455                 460                 


His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu 
465                 470                 475                 480 


Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr 
                485                 490                 495     


Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile 
            500                 505                 510         


Gly Lys Ala Thr Ala Lys Arg Phe Phe Tyr Ser Asn Ile Met Asn Phe 
        515                 520                 525             


Phe Lys Thr Glu Val Lys Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro 
    530                 535                 540                 


Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Val Val Trp Asn Lys Glu 
545                 550                 555                 560 


Lys Asp Phe Ala Thr Val Arg Lys Val Leu Ala Met Pro Gln Val Asn 
                565                 570                 575     


Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser 
            580                 585                 590         


Ile Leu Ser Lys Arg Glu Ser Ala Lys Leu Ile Pro Arg Lys Lys Gly 
        595                 600                 605             


Trp Asp Thr Arg Lys Tyr Gly Gly Phe Gly Ser Pro Thr Val Ala Tyr 
    610                 615                 620                 


Ser Ile Leu Val Val Ala Lys Val Glu Lys Gly Lys Ala Lys Lys Leu 
625                 630                 635                 640 


Lys Ser Val Lys Val Leu Val Gly Ile Thr Ile Met Glu Lys Gly Ser 
                645                 650                 655     


Tyr Glu Lys Asp Pro Ile Gly Phe Leu Glu Ala Lys Gly Tyr Lys Asp 
            660                 665                 670         


Ile Lys Lys Glu Leu Ile Phe Lys Leu Pro Lys Tyr Ser Leu Phe Glu 
        675                 680                 685             


Leu Glu Asn Gly Arg Arg Arg Met Leu Ala Ser Ala Thr Glu Leu Gln 
    690                 695                 700                 


Lys Ala Asn Glu Leu Val Leu Pro Gln His Leu Val Arg Leu Leu Tyr 
705                 710                 715                 720 


Tyr Thr Gln Asn Ile Ser Ala Thr Thr Gly Ser Asn Asn Leu Gly Tyr 
                725                 730                 735     


Ile Glu Gln His Arg Glu Glu Phe Lys Glu Ile Phe Glu Lys Ile Ile 
            740                 745                 750         


Asp Phe Ser Glu Lys Tyr Ile Leu Lys Asn Lys Val Asn Ser Asn Leu 
        755                 760                 765             


Lys Ser Ser Phe Asp Glu Gln Phe Ala Val Ser Asp Ser Ile Leu Leu 
    770                 775                 780                 


Ser Asn Ser Phe Val Ser Leu Leu Lys Tyr Thr Ser Phe Gly Ala Ser 
785                 790                 795                 800 


Gly Gly Phe Thr Phe Leu Asp Leu Asp Val Lys Gln Gly Arg Leu Arg 
                805                 810                 815     


Tyr Gln Thr Val Thr Glu Val Leu Asp Ala Thr Leu Ile Tyr Gln Ser 
            820                 825                 830         


Ile Thr Gly Leu Tyr Glu Thr Arg Thr Asp Leu Ser Gln Leu Gly Gly 
        835                 840                 845             


Asp Ser Gly Gly Ser Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser 
    850                 855                 860                 


Pro Lys Lys Lys Arg Lys Val 
865                 870     


<210>  353
<211>  932
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok Intein, N-terminal, Construct 14870

<400>  353

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn 
225                 230                 235                 240 


Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys 
                245                 250                 255     


Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys Lys Asn 
            260                 265                 270         


Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr 
        275                 280                 285             


Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg 
    290                 295                 300                 


Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys Leu Asp 
305                 310                 315                 320 


Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp 
                325                 330                 335     


Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp Glu Val 
            340                 345                 350         


Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu 
        355                 360                 365             


Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu 
    370                 375                 380                 


Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Lys Leu 
385                 390                 395                 400 


Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu Ile Gln 
                405                 410                 415     


Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile Glu Val 
            420                 425                 430         


Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys Arg Leu 
        435                 440                 445             


Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly Leu Phe 
    450                 455                 460                 


Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe Lys Ser 
465                 470                 475                 480 


Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr 
                485                 490                 495     


Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp Gln Tyr 
            500                 505                 510         


Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu 
        515                 520                 525             


Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro Leu Ser 
    530                 535                 540                 


Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu Ala Leu 
545                 550                 555                 560 


Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala Glu Ile 
                565                 570                 575     


Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly Ile Gly 
            580                 585                 590         


Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu Glu Phe 
        595                 600                 605             


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala Glu Glu 
    610                 615                 620                 


Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln Arg Thr 
625                 630                 635                 640 


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu Leu His 
                645                 650                 655     


Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys Glu Asn 
            660                 665                 670         


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
        675                 680                 685             


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr Arg Lys 
    690                 695                 700                 


Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
705                 710                 715                 720 


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Glu 
                725                 730                 735     


Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
            740                 745                 750         


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
        755                 760                 765             


Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
    770                 775                 780                 


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
785                 790                 795                 800 


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Leu Ser Tyr Glu Thr Glu 
                805                 810                 815     


Ile Leu Thr Val Glu Tyr Gly Leu Leu Pro Ile Gly Lys Ile Val Glu 
            820                 825                 830         


Lys Arg Ile Glu Cys Thr Val Tyr Ser Val Asp Asn Asn Gly Asn Ile 
        835                 840                 845             


Tyr Thr Gln Pro Val Ala Gln Trp His Asp Arg Gly Glu Gln Glu Val 
    850                 855                 860                 


Phe Glu Tyr Cys Leu Glu Asp Gly Ser Leu Ile Arg Ala Thr Lys Asp 
865                 870                 875                 880 


His Lys Phe Met Thr Val Asp Gly Gln Met Leu Pro Ile Asp Glu Ile 
                885                 890                 895     


Phe Glu Arg Glu Leu Asp Leu Met Arg Val Asp Asn Leu Pro Asn Ser 
            900                 905                 910         


Gly Gly Ser Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Pro Lys Lys 
        915                 920                 925             


Lys Arg Lys Val 
    930         


<210>  354
<211>  1700
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dScCas-FokI

<400>  354

Met Ala Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val Ser Ser 
1               5                   10                  15      


Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg Leu 
            20                  25                  30          


Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala Ala 
        35                  40                  45              


Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys Arg 
    50                  55                  60                  


Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys Glu 
65                  70                  75                  80  


Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro Ala 
                85                  90                  95      


Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly Ser Gly 
            100                 105                 110         


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
        115                 120                 125             


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
    130                 135                 140                 


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
145                 150                 155                 160 


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
                165                 170                 175     


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
            180                 185                 190         


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
        195                 200                 205             


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
    210                 215                 220                 


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
225                 230                 235                 240 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
                245                 250                 255     


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
            260                 265                 270         


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
        275                 280                 285             


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
    290                 295                 300                 


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
305                 310                 315                 320 


Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile 
                325                 330                 335     


Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val 
            340                 345                 350         


Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile 
        355                 360                 365             


Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala 
    370                 375                 380                 


Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg 
385                 390                 395                 400 


Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala 
                405                 410                 415     


Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val 
            420                 425                 430         


Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala 
        435                 440                 445             


Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg 
    450                 455                 460                 


Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr 
465                 470                 475                 480 


Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu 
                485                 490                 495     


Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln 
            500                 505                 510         


Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu 
        515                 520                 525             


Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser 
    530                 535                 540                 


Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn 
545                 550                 555                 560 


Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn 
                565                 570                 575     


Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser 
            580                 585                 590         


Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly 
        595                 600                 605             


Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala 
    610                 615                 620                 


Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala 
625                 630                 635                 640 


Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp 
                645                 650                 655     


Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr 
            660                 665                 670         


Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val 
        675                 680                 685             


Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln 
    690                 695                 700                 


Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
705                 710                 715                 720 


Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys 
                725                 730                 735     


Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys 
            740                 745                 750         


Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu 
        755                 760                 765             


Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
    770                 775                 780                 


Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu 
785                 790                 795                 800 


Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val 
                805                 810                 815     


Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
            820                 825                 830         


Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu 
        835                 840                 845             


Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
    850                 855                 860                 


Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys 
865                 870                 875                 880 


Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
                885                 890                 895     


Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
            900                 905                 910         


Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
        915                 920                 925             


Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
    930                 935                 940                 


Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
945                 950                 955                 960 


Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
                965                 970                 975     


Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr 
            980                 985                 990         


Gly Trp Gly Arg Leu Ser Arg Lys  Met Ile Asn Gly Ile  Arg Asp Lys 
        995                 1000                 1005             


Gln Ser  Gly Lys Thr Ile Leu  Asp Phe Leu Lys Ser  Asp Gly Phe 
    1010                 1015                 1020             


Ser Asn  Arg Asn Phe Met Gln  Leu Ile His Asp Asp  Ser Leu Thr 
    1025                 1030                 1035             


Phe Lys  Glu Glu Ile Glu Lys  Ala Gln Val Ser Gly  Gln Gly Asp 
    1040                 1045                 1050             


Ser Leu  His Glu Gln Ile Ala  Asp Leu Ala Gly Ser  Pro Ala Ile 
    1055                 1060                 1065             


Lys Lys  Gly Ile Leu Gln Thr  Val Lys Ile Val Asp  Glu Leu Val 
    1070                 1075                 1080             


Lys Val  Met Gly His Lys Pro  Glu Asn Ile Val Ile  Glu Met Ala 
    1085                 1090                 1095             


Arg Glu  Asn Gln Thr Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu 
    1100                 1105                 1110             


Arg Lys  Lys Arg Ile Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln 
    1115                 1120                 1125             


Ile Leu  Lys Glu Asn Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
    1130                 1135                 1140             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1145                 1150                 1155             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1160                 1165                 1170             


Ala Ile  Val Pro Gln Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn 
    1175                 1180                 1185             


Lys Val  Leu Thr Arg Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn 
    1190                 1195                 1200             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1205                 1210                 1215             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1220                 1225                 1230             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala 
    1235                 1240                 1245             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1250                 1255                 1260             


His Val  Ala Arg Ile Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp 
    1265                 1270                 1275             


Lys Asn  Asp Lys Pro Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1280                 1285                 1290             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys 
    1295                 1300                 1305             


Val Arg  Asp Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1310                 1315                 1320             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1325                 1330                 1335             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1340                 1345                 1350             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1355                 1360                 1365             


Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1370                 1375                 1380             


Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1385                 1390                 1395             


Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp 
    1400                 1405                 1410             


Phe Ala  Thr Val Arg Lys Val  Leu Ala Met Pro Gln  Val Asn Ile 
    1415                 1420                 1425             


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1430                 1435                 1440             


Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys 
    1445                 1450                 1455             


Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val 
    1460                 1465                 1470             


Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ala 
    1475                 1480                 1485             


Lys Lys  Leu Lys Ser Val Lys  Val Leu Val Gly Ile  Thr Ile Met 
    1490                 1495                 1500             


Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala 
    1505                 1510                 1515             


Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro 
    1520                 1525                 1530             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu 
    1535                 1540                 1545             


Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro 
    1550                 1555                 1560             


Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala 
    1565                 1570                 1575             


Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu 
    1580                 1585                 1590             


Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys 
    1595                 1600                 1605             


Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe 
    1610                 1615                 1620             


Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser 
    1625                 1630                 1635             


Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly 
    1640                 1645                 1650             


Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr 
    1655                 1660                 1665             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1670                 1675                 1680             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1685                 1690                 1695             


Gly Asp  
    1700 


<210>  355
<211>  1783
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dScCas-FokI-HMGB1

<400>  355

Met Ala Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val Ser Ser 
1               5                   10                  15      


Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg Leu 
            20                  25                  30          


Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala Ala 
        35                  40                  45              


Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys Arg 
    50                  55                  60                  


Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys Glu 
65                  70                  75                  80  


Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro Ala 
                85                  90                  95      


Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly Ser Gly 
            100                 105                 110         


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
        115                 120                 125             


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
    130                 135                 140                 


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
145                 150                 155                 160 


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
                165                 170                 175     


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
            180                 185                 190         


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
        195                 200                 205             


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
    210                 215                 220                 


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
225                 230                 235                 240 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
                245                 250                 255     


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
            260                 265                 270         


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
        275                 280                 285             


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
    290                 295                 300                 


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
305                 310                 315                 320 


Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile 
                325                 330                 335     


Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val 
            340                 345                 350         


Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile 
        355                 360                 365             


Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala 
    370                 375                 380                 


Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg 
385                 390                 395                 400 


Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala 
                405                 410                 415     


Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val 
            420                 425                 430         


Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala 
        435                 440                 445             


Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg 
    450                 455                 460                 


Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr 
465                 470                 475                 480 


Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu 
                485                 490                 495     


Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln 
            500                 505                 510         


Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu 
        515                 520                 525             


Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser 
    530                 535                 540                 


Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn 
545                 550                 555                 560 


Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn 
                565                 570                 575     


Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser 
            580                 585                 590         


Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly 
        595                 600                 605             


Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala 
    610                 615                 620                 


Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala 
625                 630                 635                 640 


Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp 
                645                 650                 655     


Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr 
            660                 665                 670         


Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val 
        675                 680                 685             


Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln 
    690                 695                 700                 


Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
705                 710                 715                 720 


Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys 
                725                 730                 735     


Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys 
            740                 745                 750         


Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu 
        755                 760                 765             


Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
    770                 775                 780                 


Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu 
785                 790                 795                 800 


Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val 
                805                 810                 815     


Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
            820                 825                 830         


Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu 
        835                 840                 845             


Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
    850                 855                 860                 


Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys 
865                 870                 875                 880 


Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
                885                 890                 895     


Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
            900                 905                 910         


Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
        915                 920                 925             


Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
    930                 935                 940                 


Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
945                 950                 955                 960 


Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
                965                 970                 975     


Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr 
            980                 985                 990         


Gly Trp Gly Arg Leu Ser Arg Lys  Met Ile Asn Gly Ile  Arg Asp Lys 
        995                 1000                 1005             


Gln Ser  Gly Lys Thr Ile Leu  Asp Phe Leu Lys Ser  Asp Gly Phe 
    1010                 1015                 1020             


Ser Asn  Arg Asn Phe Met Gln  Leu Ile His Asp Asp  Ser Leu Thr 
    1025                 1030                 1035             


Phe Lys  Glu Glu Ile Glu Lys  Ala Gln Val Ser Gly  Gln Gly Asp 
    1040                 1045                 1050             


Ser Leu  His Glu Gln Ile Ala  Asp Leu Ala Gly Ser  Pro Ala Ile 
    1055                 1060                 1065             


Lys Lys  Gly Ile Leu Gln Thr  Val Lys Ile Val Asp  Glu Leu Val 
    1070                 1075                 1080             


Lys Val  Met Gly His Lys Pro  Glu Asn Ile Val Ile  Glu Met Ala 
    1085                 1090                 1095             


Arg Glu  Asn Gln Thr Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu 
    1100                 1105                 1110             


Arg Lys  Lys Arg Ile Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln 
    1115                 1120                 1125             


Ile Leu  Lys Glu Asn Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
    1130                 1135                 1140             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1145                 1150                 1155             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1160                 1165                 1170             


Ala Ile  Val Pro Gln Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn 
    1175                 1180                 1185             


Lys Val  Leu Thr Arg Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn 
    1190                 1195                 1200             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1205                 1210                 1215             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1220                 1225                 1230             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala 
    1235                 1240                 1245             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1250                 1255                 1260             


His Val  Ala Arg Ile Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp 
    1265                 1270                 1275             


Lys Asn  Asp Lys Pro Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1280                 1285                 1290             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys 
    1295                 1300                 1305             


Val Arg  Asp Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1310                 1315                 1320             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1325                 1330                 1335             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1340                 1345                 1350             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1355                 1360                 1365             


Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1370                 1375                 1380             


Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1385                 1390                 1395             


Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp 
    1400                 1405                 1410             


Phe Ala  Thr Val Arg Lys Val  Leu Ala Met Pro Gln  Val Asn Ile 
    1415                 1420                 1425             


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1430                 1435                 1440             


Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys 
    1445                 1450                 1455             


Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val 
    1460                 1465                 1470             


Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ala 
    1475                 1480                 1485             


Lys Lys  Leu Lys Ser Val Lys  Val Leu Val Gly Ile  Thr Ile Met 
    1490                 1495                 1500             


Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala 
    1505                 1510                 1515             


Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro 
    1520                 1525                 1530             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu 
    1535                 1540                 1545             


Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro 
    1550                 1555                 1560             


Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala 
    1565                 1570                 1575             


Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu 
    1580                 1585                 1590             


Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys 
    1595                 1600                 1605             


Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe 
    1610                 1615                 1620             


Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser 
    1625                 1630                 1635             


Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly 
    1640                 1645                 1650             


Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr 
    1655                 1660                 1665             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1670                 1675                 1680             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1685                 1690                 1695             


Gly Asp  Gly Lys Gly Asp Pro  Lys Lys Pro Arg Gly  Lys Met Ser 
    1700                 1705                 1710             


Ser Tyr  Ala Phe Phe Val Gln  Thr Cys Arg Glu Glu  His Lys Lys 
    1715                 1720                 1725             


Lys His  Pro Asp Ala Ser Val  Asn Phe Ser Glu Phe  Ser Lys Lys 
    1730                 1735                 1740             


Cys Ser  Glu Arg Trp Lys Thr  Met Ser Ala Lys Glu  Lys Gly Lys 
    1745                 1750                 1755             


Phe Glu  Asp Met Ala Lys Ala  Asp Lys Ala Arg Tyr  Glu Arg Glu 
    1760                 1765                 1770             


Met Lys  Thr Tyr Ile Pro Pro  Lys Gly Glu 
    1775                 1780             


<210>  356
<211>  1786
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dScCas-FokI-HMGB3

<400>  356

Met Ala Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val Ser Ser 
1               5                   10                  15      


Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg Leu 
            20                  25                  30          


Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala Ala 
        35                  40                  45              


Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys Arg 
    50                  55                  60                  


Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys Glu 
65                  70                  75                  80  


Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro Ala 
                85                  90                  95      


Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly Ser Gly 
            100                 105                 110         


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
        115                 120                 125             


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
    130                 135                 140                 


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
145                 150                 155                 160 


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
                165                 170                 175     


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
            180                 185                 190         


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
        195                 200                 205             


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
    210                 215                 220                 


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
225                 230                 235                 240 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
                245                 250                 255     


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
            260                 265                 270         


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
        275                 280                 285             


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
    290                 295                 300                 


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
305                 310                 315                 320 


Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile 
                325                 330                 335     


Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val 
            340                 345                 350         


Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile 
        355                 360                 365             


Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala 
    370                 375                 380                 


Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg 
385                 390                 395                 400 


Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala 
                405                 410                 415     


Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val 
            420                 425                 430         


Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala 
        435                 440                 445             


Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg 
    450                 455                 460                 


Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr 
465                 470                 475                 480 


Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu 
                485                 490                 495     


Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln 
            500                 505                 510         


Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu 
        515                 520                 525             


Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser 
    530                 535                 540                 


Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn 
545                 550                 555                 560 


Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn 
                565                 570                 575     


Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser 
            580                 585                 590         


Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly 
        595                 600                 605             


Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala 
    610                 615                 620                 


Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala 
625                 630                 635                 640 


Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp 
                645                 650                 655     


Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr 
            660                 665                 670         


Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val 
        675                 680                 685             


Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln 
    690                 695                 700                 


Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
705                 710                 715                 720 


Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys 
                725                 730                 735     


Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys 
            740                 745                 750         


Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu 
        755                 760                 765             


Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
    770                 775                 780                 


Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu 
785                 790                 795                 800 


Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val 
                805                 810                 815     


Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
            820                 825                 830         


Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu 
        835                 840                 845             


Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
    850                 855                 860                 


Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys 
865                 870                 875                 880 


Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
                885                 890                 895     


Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
            900                 905                 910         


Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
        915                 920                 925             


Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
    930                 935                 940                 


Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
945                 950                 955                 960 


Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
                965                 970                 975     


Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr 
            980                 985                 990         


Gly Trp Gly Arg Leu Ser Arg Lys  Met Ile Asn Gly Ile  Arg Asp Lys 
        995                 1000                 1005             


Gln Ser  Gly Lys Thr Ile Leu  Asp Phe Leu Lys Ser  Asp Gly Phe 
    1010                 1015                 1020             


Ser Asn  Arg Asn Phe Met Gln  Leu Ile His Asp Asp  Ser Leu Thr 
    1025                 1030                 1035             


Phe Lys  Glu Glu Ile Glu Lys  Ala Gln Val Ser Gly  Gln Gly Asp 
    1040                 1045                 1050             


Ser Leu  His Glu Gln Ile Ala  Asp Leu Ala Gly Ser  Pro Ala Ile 
    1055                 1060                 1065             


Lys Lys  Gly Ile Leu Gln Thr  Val Lys Ile Val Asp  Glu Leu Val 
    1070                 1075                 1080             


Lys Val  Met Gly His Lys Pro  Glu Asn Ile Val Ile  Glu Met Ala 
    1085                 1090                 1095             


Arg Glu  Asn Gln Thr Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu 
    1100                 1105                 1110             


Arg Lys  Lys Arg Ile Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln 
    1115                 1120                 1125             


Ile Leu  Lys Glu Asn Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
    1130                 1135                 1140             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1145                 1150                 1155             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1160                 1165                 1170             


Ala Ile  Val Pro Gln Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn 
    1175                 1180                 1185             


Lys Val  Leu Thr Arg Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn 
    1190                 1195                 1200             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1205                 1210                 1215             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1220                 1225                 1230             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala 
    1235                 1240                 1245             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1250                 1255                 1260             


His Val  Ala Arg Ile Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp 
    1265                 1270                 1275             


Lys Asn  Asp Lys Pro Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1280                 1285                 1290             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys 
    1295                 1300                 1305             


Val Arg  Asp Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1310                 1315                 1320             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1325                 1330                 1335             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1340                 1345                 1350             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1355                 1360                 1365             


Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1370                 1375                 1380             


Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1385                 1390                 1395             


Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp 
    1400                 1405                 1410             


Phe Ala  Thr Val Arg Lys Val  Leu Ala Met Pro Gln  Val Asn Ile 
    1415                 1420                 1425             


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1430                 1435                 1440             


Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys 
    1445                 1450                 1455             


Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val 
    1460                 1465                 1470             


Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ala 
    1475                 1480                 1485             


Lys Lys  Leu Lys Ser Val Lys  Val Leu Val Gly Ile  Thr Ile Met 
    1490                 1495                 1500             


Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala 
    1505                 1510                 1515             


Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro 
    1520                 1525                 1530             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu 
    1535                 1540                 1545             


Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro 
    1550                 1555                 1560             


Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala 
    1565                 1570                 1575             


Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu 
    1580                 1585                 1590             


Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys 
    1595                 1600                 1605             


Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe 
    1610                 1615                 1620             


Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser 
    1625                 1630                 1635             


Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly 
    1640                 1645                 1650             


Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr 
    1655                 1660                 1665             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1670                 1675                 1680             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1685                 1690                 1695             


Gly Asp  Gly Ser Ser Gly Ser  Ser Gly Met Ala Lys  Gly Asp Pro 
    1700                 1705                 1710             


Lys Lys  Pro Lys Gly Lys Met  Ser Ala Tyr Ala Phe  Phe Val Gln 
    1715                 1720                 1725             


Thr Cys  Arg Glu Glu His Lys  Lys Lys Asn Pro Glu  Val Pro Val 
    1730                 1735                 1740             


Asn Phe  Ala Glu Phe Ser Lys  Lys Cys Ser Glu Arg  Trp Lys Thr 
    1745                 1750                 1755             


Met Ser  Gly Lys Glu Lys Ser  Lys Phe Asp Glu Met  Ala Lys Ala 
    1760                 1765                 1770             


Asp Lys  Val Arg Tyr Asp Arg  Glu Met Lys Asp Tyr  Gly 
    1775                 1780                 1785     


<210>  357
<211>  1782
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HMGN-dScCas-FokI-HMGB4

<400>  357

Met Ala Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val Ser Ser 
1               5                   10                  15      


Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala Arg Leu 
            20                  25                  30          


Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys Ala Ala 
        35                  40                  45              


Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly Lys Arg 
    50                  55                  60                  


Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr Lys Glu 
65                  70                  75                  80  


Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser Pro Ala 
                85                  90                  95      


Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly Ser Gly 
            100                 105                 110         


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
        115                 120                 125             


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
    130                 135                 140                 


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
145                 150                 155                 160 


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
                165                 170                 175     


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
            180                 185                 190         


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
        195                 200                 205             


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
    210                 215                 220                 


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
225                 230                 235                 240 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
                245                 250                 255     


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
            260                 265                 270         


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
        275                 280                 285             


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
    290                 295                 300                 


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
305                 310                 315                 320 


Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile 
                325                 330                 335     


Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val 
            340                 345                 350         


Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile 
        355                 360                 365             


Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala 
    370                 375                 380                 


Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg 
385                 390                 395                 400 


Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala 
                405                 410                 415     


Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val 
            420                 425                 430         


Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala 
        435                 440                 445             


Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg 
    450                 455                 460                 


Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr 
465                 470                 475                 480 


Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu 
                485                 490                 495     


Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln 
            500                 505                 510         


Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu 
        515                 520                 525             


Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser 
    530                 535                 540                 


Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn 
545                 550                 555                 560 


Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn 
                565                 570                 575     


Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser 
            580                 585                 590         


Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly 
        595                 600                 605             


Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala 
    610                 615                 620                 


Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala 
625                 630                 635                 640 


Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp 
                645                 650                 655     


Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr 
            660                 665                 670         


Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val 
        675                 680                 685             


Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln 
    690                 695                 700                 


Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
705                 710                 715                 720 


Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys 
                725                 730                 735     


Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys 
            740                 745                 750         


Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu 
        755                 760                 765             


Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
    770                 775                 780                 


Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu 
785                 790                 795                 800 


Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val 
                805                 810                 815     


Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
            820                 825                 830         


Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu 
        835                 840                 845             


Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
    850                 855                 860                 


Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys 
865                 870                 875                 880 


Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
                885                 890                 895     


Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
            900                 905                 910         


Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
        915                 920                 925             


Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
    930                 935                 940                 


Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
945                 950                 955                 960 


Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
                965                 970                 975     


Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr 
            980                 985                 990         


Gly Trp Gly Arg Leu Ser Arg Lys  Met Ile Asn Gly Ile  Arg Asp Lys 
        995                 1000                 1005             


Gln Ser  Gly Lys Thr Ile Leu  Asp Phe Leu Lys Ser  Asp Gly Phe 
    1010                 1015                 1020             


Ser Asn  Arg Asn Phe Met Gln  Leu Ile His Asp Asp  Ser Leu Thr 
    1025                 1030                 1035             


Phe Lys  Glu Glu Ile Glu Lys  Ala Gln Val Ser Gly  Gln Gly Asp 
    1040                 1045                 1050             


Ser Leu  His Glu Gln Ile Ala  Asp Leu Ala Gly Ser  Pro Ala Ile 
    1055                 1060                 1065             


Lys Lys  Gly Ile Leu Gln Thr  Val Lys Ile Val Asp  Glu Leu Val 
    1070                 1075                 1080             


Lys Val  Met Gly His Lys Pro  Glu Asn Ile Val Ile  Glu Met Ala 
    1085                 1090                 1095             


Arg Glu  Asn Gln Thr Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu 
    1100                 1105                 1110             


Arg Lys  Lys Arg Ile Glu Glu  Gly Ile Lys Glu Leu  Glu Ser Gln 
    1115                 1120                 1125             


Ile Leu  Lys Glu Asn Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
    1130                 1135                 1140             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1145                 1150                 1155             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1160                 1165                 1170             


Ala Ile  Val Pro Gln Ser Phe  Ile Lys Asp Asp Ser  Ile Asp Asn 
    1175                 1180                 1185             


Lys Val  Leu Thr Arg Ser Val  Glu Asn Arg Gly Lys  Ser Asp Asn 
    1190                 1195                 1200             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1205                 1210                 1215             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1220                 1225                 1230             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Ala  Asp Lys Ala 
    1235                 1240                 1245             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1250                 1255                 1260             


His Val  Ala Arg Ile Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp 
    1265                 1270                 1275             


Lys Asn  Asp Lys Pro Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1280                 1285                 1290             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys 
    1295                 1300                 1305             


Val Arg  Asp Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1310                 1315                 1320             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1325                 1330                 1335             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1340                 1345                 1350             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1355                 1360                 1365             


Lys Arg  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1370                 1375                 1380             


Val Lys  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1385                 1390                 1395             


Thr Asn  Gly Glu Thr Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp 
    1400                 1405                 1410             


Phe Ala  Thr Val Arg Lys Val  Leu Ala Met Pro Gln  Val Asn Ile 
    1415                 1420                 1425             


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1430                 1435                 1440             


Ile Leu  Ser Lys Arg Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys 
    1445                 1450                 1455             


Gly Trp  Asp Thr Arg Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val 
    1460                 1465                 1470             


Ala Tyr  Ser Ile Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ala 
    1475                 1480                 1485             


Lys Lys  Leu Lys Ser Val Lys  Val Leu Val Gly Ile  Thr Ile Met 
    1490                 1495                 1500             


Glu Lys  Gly Ser Tyr Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala 
    1505                 1510                 1515             


Lys Gly  Tyr Lys Asp Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro 
    1520                 1525                 1530             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu 
    1535                 1540                 1545             


Ala Ser  Ala Thr Glu Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro 
    1550                 1555                 1560             


Gln His  Leu Val Arg Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala 
    1565                 1570                 1575             


Thr Thr  Gly Ser Asn Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu 
    1580                 1585                 1590             


Glu Phe  Lys Glu Ile Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys 
    1595                 1600                 1605             


Tyr Ile  Leu Lys Asn Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe 
    1610                 1615                 1620             


Asp Glu  Gln Phe Ala Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser 
    1625                 1630                 1635             


Phe Val  Ser Leu Leu Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly 
    1640                 1645                 1650             


Phe Thr  Phe Leu Asp Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr 
    1655                 1660                 1665             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1670                 1675                 1680             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1685                 1690                 1695             


Gly Asp  Gly Gly Ser Gly Gly  Lys Glu Ile Gln Leu  Lys Pro Lys 
    1700                 1705                 1710             


Ala Asn  Val Ser Ser Tyr Val  His Phe Leu Leu Asn  Tyr Arg Asn 
    1715                 1720                 1725             


Lys Phe  Lys Glu Gln Gln Pro  Asn Thr Tyr Val Gly  Phe Lys Glu 
    1730                 1735                 1740             


Phe Ser  Arg Lys Cys Ser Glu  Lys Trp Arg Ser Ile  Ser Lys His 
    1745                 1750                 1755             


Glu Lys  Ala Lys Tyr Glu Ala  Leu Ala Lys Leu Asp  Lys Ala Arg 
    1760                 1765                 1770             


Tyr Gln  Glu Glu Met Met Asn  Tyr Val 
    1775                 1780         


<210>  358
<211>  707
<212>  PRT
<213>  Biggiephage clade


<220>
<221>  MISC_FEATURE
<223>  CasF-1

<400>  358

Met Ala Asp Thr Pro Thr Leu Phe Thr Gln Phe Leu Arg His His Leu 
1               5                   10                  15      


Pro Gly Gln Arg Phe Arg Lys Asp Ile Leu Lys Gln Ala Gly Arg Ile 
            20                  25                  30          


Leu Ala Asn Lys Gly Glu Asp Ala Thr Ile Ala Phe Leu Arg Gly Lys 
        35                  40                  45              


Ser Glu Glu Ser Pro Pro Asp Phe Gln Pro Pro Val Lys Cys Pro Ile 
    50                  55                  60                  


Ile Ala Cys Ser Arg Pro Leu Thr Glu Trp Pro Ile Tyr Gln Ala Ser 
65                  70                  75                  80  


Val Ala Ile Gln Gly Tyr Val Tyr Gly Gln Ser Leu Ala Glu Phe Glu 
                85                  90                  95      


Ala Ser Asp Pro Gly Cys Ser Lys Asp Gly Leu Leu Gly Trp Phe Asp 
            100                 105                 110         


Lys Thr Gly Val Cys Thr Asp Tyr Phe Ser Val Gln Gly Leu Asn Leu 
        115                 120                 125             


Ile Phe Gln Asn Ala Arg Lys Arg Tyr Ile Gly Val Gln Thr Lys Val 
    130                 135                 140                 


Thr Asn Arg Asn Glu Lys Arg His Lys Lys Leu Lys Arg Ile Asn Ala 
145                 150                 155                 160 


Lys Arg Ile Ala Glu Gly Leu Pro Glu Leu Thr Ser Asp Glu Pro Glu 
                165                 170                 175     


Ser Ala Leu Asp Glu Thr Gly His Leu Ile Asp Pro Pro Gly Leu Asn 
            180                 185                 190         


Thr Asn Ile Tyr Cys Tyr Gln Gln Val Ser Pro Lys Pro Leu Ala Leu 
        195                 200                 205             


Ser Glu Val Asn Gln Leu Pro Thr Ala Tyr Ala Gly Tyr Ser Thr Ser 
    210                 215                 220                 


Gly Asp Asp Pro Ile Gln Pro Met Val Thr Lys Asp Arg Leu Ser Ile 
225                 230                 235                 240 


Ser Lys Gly Gln Pro Gly Tyr Ile Pro Glu His Gln Arg Ala Leu Leu 
                245                 250                 255     


Ser Gln Lys Lys His Arg Arg Met Arg Gly Tyr Gly Leu Lys Ala Arg 
            260                 265                 270         


Ala Leu Leu Val Ile Val Arg Ile Gln Asp Asp Trp Ala Val Ile Asp 
        275                 280                 285             


Leu Arg Ser Leu Leu Arg Asn Ala Tyr Trp Arg Arg Ile Val Gln Thr 
    290                 295                 300                 


Lys Glu Pro Ser Thr Ile Thr Lys Leu Leu Lys Leu Val Thr Gly Asp 
305                 310                 315                 320 


Pro Val Leu Asp Ala Thr Arg Met Val Ala Thr Phe Thr Tyr Lys Pro 
                325                 330                 335     


Gly Ile Val Gln Val Arg Ser Ala Lys Cys Leu Lys Asn Lys Gln Gly 
            340                 345                 350         


Ser Lys Leu Phe Ser Glu Arg Tyr Leu Asn Glu Thr Val Ser Val Thr 
        355                 360                 365             


Ser Ile Asp Leu Gly Ser Asn Asn Leu Val Ala Val Ala Thr Tyr Arg 
    370                 375                 380                 


Leu Val Asn Gly Asn Thr Pro Glu Leu Leu Gln Arg Phe Thr Leu Pro 
385                 390                 395                 400 


Ser His Leu Val Lys Asp Phe Glu Arg Tyr Lys Gln Ala His Asp Thr 
                405                 410                 415     


Leu Glu Asp Ser Ile Gln Lys Thr Ala Val Ala Ser Leu Pro Gln Gly 
            420                 425                 430         


Gln Gln Thr Glu Ile Arg Met Trp Ser Met Tyr Gly Phe Arg Glu Ala 
        435                 440                 445             


Gln Glu Arg Val Cys Gln Glu Leu Gly Leu Ala Asp Gly Ser Ile Pro 
    450                 455                 460                 


Trp Asn Val Met Thr Ala Thr Ser Thr Ile Leu Thr Asp Leu Phe Leu 
465                 470                 475                 480 


Ala Arg Gly Gly Asp Pro Lys Lys Cys Met Phe Thr Ser Glu Pro Lys 
                485                 490                 495     


Lys Lys Lys Asn Ser Lys Gln Val Leu Tyr Lys Ile Arg Asp Arg Ala 
            500                 505                 510         


Trp Ala Lys Met Tyr Arg Thr Leu Leu Ser Lys Glu Thr Arg Glu Ala 
        515                 520                 525             


Trp Asn Lys Ala Leu Trp Gly Leu Lys Arg Gly Ser Pro Asp Tyr Ala 
    530                 535                 540                 


Arg Leu Ser Lys Arg Lys Glu Glu Leu Ala Arg Arg Cys Val Asn Tyr 
545                 550                 555                 560 


Thr Ile Ser Thr Ala Glu Lys Arg Ala Gln Cys Gly Arg Thr Ile Val 
                565                 570                 575     


Ala Leu Glu Asp Leu Asn Ile Gly Phe Phe His Gly Arg Gly Lys Gln 
            580                 585                 590         


Glu Pro Gly Trp Val Gly Leu Phe Thr Arg Lys Lys Glu Asn Arg Trp 
        595                 600                 605             


Leu Met Gln Ala Leu His Lys Ala Phe Leu Glu Leu Ala His His Arg 
    610                 615                 620                 


Gly Tyr His Val Ile Glu Val Asn Pro Ala Tyr Thr Ser Gln Thr Cys 
625                 630                 635                 640 


Pro Val Cys Arg His Cys Asp Pro Asp Asn Arg Asp Gln His Asn Arg 
                645                 650                 655     


Glu Ala Phe His Cys Ile Gly Cys Gly Phe Arg Gly Asn Ala Asp Leu 
            660                 665                 670         


Asp Val Ala Thr His Asn Ile Ala Met Val Ala Ile Thr Gly Glu Ser 
        675                 680                 685             


Leu Lys Arg Ala Arg Gly Ser Val Ala Ser Lys Thr Pro Gln Pro Leu 
    690                 695                 700                 


Ala Ala Glu 
705         


<210>  359
<211>  757
<212>  PRT
<213>  Biggiephage clade


<220>
<221>  MISC_FEATURE
<223>  CasF-2

<400>  359

Met Pro Lys Pro Ala Val Glu Ser Glu Phe Ser Lys Val Leu Lys Lys 
1               5                   10                  15      


His Phe Pro Gly Glu Arg Phe Arg Ser Ser Tyr Met Lys Arg Gly Gly 
            20                  25                  30          


Lys Ile Leu Ala Ala Gln Gly Glu Glu Ala Val Val Ala Tyr Leu Gln 
        35                  40                  45              


Gly Lys Ser Glu Glu Glu Pro Pro Asn Phe Gln Pro Pro Ala Lys Cys 
    50                  55                  60                  


His Val Val Thr Lys Ser Arg Asp Phe Ala Glu Trp Pro Ile Met Lys 
65                  70                  75                  80  


Ala Ser Glu Ala Ile Gln Arg Tyr Ile Tyr Ala Leu Ser Thr Thr Glu 
                85                  90                  95      


Arg Ala Ala Cys Lys Pro Gly Lys Ser Ser Glu Ser His Ala Ala Trp 
            100                 105                 110         


Phe Ala Ala Thr Gly Val Ser Asn His Gly Tyr Ser His Val Gln Gly 
        115                 120                 125             


Leu Asn Leu Ile Phe Asp His Thr Leu Gly Arg Tyr Asp Gly Val Leu 
    130                 135                 140                 


Lys Lys Val Gln Leu Arg Asn Glu Lys Ala Arg Ala Arg Leu Glu Ser 
145                 150                 155                 160 


Ile Asn Ala Ser Arg Ala Asp Glu Gly Leu Pro Glu Ile Lys Ala Glu 
                165                 170                 175     


Glu Glu Glu Val Ala Thr Asn Glu Thr Gly His Leu Leu Gln Pro Pro 
            180                 185                 190         


Gly Ile Asn Pro Ser Phe Tyr Val Tyr Gln Thr Ile Ser Pro Gln Ala 
        195                 200                 205             


Tyr Arg Pro Arg Asp Glu Ile Val Leu Pro Pro Glu Tyr Ala Gly Tyr 
    210                 215                 220                 


Val Arg Asp Pro Asn Ala Pro Ile Pro Leu Gly Val Val Arg Asn Arg 
225                 230                 235                 240 


Cys Asp Ile Gln Lys Gly Cys Pro Gly Tyr Ile Pro Glu Trp Gln Arg 
                245                 250                 255     


Glu Ala Gly Thr Ala Ile Ser Pro Lys Thr Gly Lys Ala Val Thr Val 
            260                 265                 270         


Pro Gly Leu Ser Pro Lys Lys Asn Lys Arg Met Arg Arg Tyr Trp Arg 
        275                 280                 285             


Ser Glu Lys Glu Lys Ala Gln Asp Ala Leu Leu Val Thr Val Arg Ile 
    290                 295                 300                 


Gly Thr Asp Trp Val Val Ile Asp Val Arg Gly Leu Leu Arg Asn Ala 
305                 310                 315                 320 


Arg Trp Arg Thr Ile Ala Pro Lys Asp Ile Ser Leu Asn Ala Leu Leu 
                325                 330                 335     


Asp Leu Phe Thr Gly Asp Pro Val Ile Asp Val Arg Arg Asn Ile Val 
            340                 345                 350         


Thr Phe Thr Tyr Thr Leu Asp Ala Cys Gly Thr Tyr Ala Arg Lys Trp 
        355                 360                 365             


Thr Leu Lys Gly Lys Gln Thr Lys Ala Thr Leu Asp Lys Leu Thr Ala 
    370                 375                 380                 


Thr Gln Thr Val Ala Leu Val Ala Ile Asp Leu Gly Gln Thr Asn Pro 
385                 390                 395                 400 


Ile Ser Ala Gly Ile Ser Arg Val Thr Gln Glu Asn Gly Ala Leu Gln 
                405                 410                 415     


Cys Glu Pro Leu Asp Arg Phe Thr Leu Pro Asp Asp Leu Leu Lys Asp 
            420                 425                 430         


Ile Ser Ala Tyr Arg Ile Ala Trp Asp Arg Asn Glu Glu Glu Leu Arg 
        435                 440                 445             


Ala Arg Ser Val Glu Ala Leu Pro Glu Ala Gln Gln Ala Glu Val Arg 
    450                 455                 460                 


Ala Leu Asp Gly Val Ser Lys Glu Thr Ala Arg Thr Gln Leu Cys Ala 
465                 470                 475                 480 


Asp Phe Gly Leu Asp Pro Lys Arg Leu Pro Trp Asp Lys Met Ser Ser 
                485                 490                 495     


Asn Thr Thr Phe Ile Ser Glu Ala Leu Leu Ser Asn Ser Val Ser Arg 
            500                 505                 510         


Asp Gln Val Phe Phe Thr Pro Ala Pro Lys Lys Gly Ala Lys Lys Lys 
        515                 520                 525             


Ala Pro Val Glu Val Met Arg Lys Asp Arg Thr Trp Ala Arg Ala Tyr 
    530                 535                 540                 


Lys Pro Arg Leu Ser Val Glu Ala Gln Lys Leu Lys Asn Glu Ala Leu 
545                 550                 555                 560 


Trp Ala Leu Lys Arg Thr Ser Pro Glu Tyr Leu Lys Leu Ser Arg Arg 
                565                 570                 575     


Lys Glu Glu Leu Cys Arg Arg Ser Ile Asn Tyr Val Ile Glu Lys Thr 
            580                 585                 590         


Arg Arg Arg Thr Gln Cys Gln Ile Val Ile Pro Val Ile Glu Asp Leu 
        595                 600                 605             


Asn Val Arg Phe Phe His Gly Ser Gly Lys Arg Leu Pro Gly Trp Asp 
    610                 615                 620                 


Asn Phe Phe Thr Ala Lys Lys Glu Asn Arg Trp Phe Ile Gln Gly Leu 
625                 630                 635                 640 


His Lys Ala Phe Ser Asp Leu Arg Thr His Arg Ser Phe Tyr Val Phe 
                645                 650                 655     


Glu Val Arg Pro Glu Arg Thr Ser Ile Thr Cys Pro Lys Cys Gly His 
            660                 665                 670         


Cys Glu Val Gly Asn Arg Asp Gly Glu Ala Phe Gln Cys Leu Ser Cys 
        675                 680                 685             


Gly Lys Thr Cys Asn Ala Asp Leu Asp Val Ala Thr His Asn Leu Thr 
    690                 695                 700                 


Gln Val Ala Leu Thr Gly Lys Thr Met Pro Lys Arg Glu Glu Pro Arg 
705                 710                 715                 720 


Asp Ala Gln Gly Thr Ala Pro Ala Arg Lys Thr Lys Lys Ala Ser Lys 
                725                 730                 735     


Ser Lys Ala Pro Pro Ala Glu Arg Glu Asp Gln Thr Pro Ala Gln Glu 
            740                 745                 750         


Pro Ser Gln Thr Ser 
        755         


<210>  360
<211>  766
<212>  PRT
<213>  Biggiephage clade


<220>
<221>  MISC_FEATURE
<223>  CasF-3

<400>  360

Met Glu Lys Glu Ile Thr Glu Leu Thr Lys Ile Arg Arg Glu Phe Pro 
1               5                   10                  15      


Asn Lys Lys Phe Ser Ser Thr Asp Met Lys Lys Ala Gly Lys Leu Leu 
            20                  25                  30          


Lys Ala Glu Gly Pro Asp Ala Val Arg Asp Phe Leu Asn Ser Cys Gln 
        35                  40                  45              


Glu Ile Ile Gly Asp Phe Lys Pro Pro Val Lys Thr Asn Ile Val Ser 
    50                  55                  60                  


Ile Ser Arg Pro Phe Glu Glu Trp Pro Val Ser Met Val Gly Arg Ala 
65                  70                  75                  80  


Ile Gln Glu Tyr Tyr Phe Ser Leu Thr Lys Glu Glu Leu Glu Ser Val 
                85                  90                  95      


His Pro Gly Thr Ser Ser Glu Asp His Lys Ser Phe Phe Asn Ile Thr 
            100                 105                 110         


Gly Leu Ser Asn Tyr Asn Tyr Thr Ser Val Gln Gly Leu Asn Leu Ile 
        115                 120                 125             


Phe Lys Asn Ala Lys Ala Ile Tyr Asp Gly Thr Leu Val Lys Ala Asn 
    130                 135                 140                 


Asn Lys Asn Lys Lys Leu Glu Lys Lys Phe Asn Glu Ile Asn His Lys 
145                 150                 155                 160 


Arg Ser Leu Glu Gly Leu Pro Ile Ile Thr Pro Asp Phe Glu Glu Pro 
                165                 170                 175     


Phe Asp Glu Asn Gly His Leu Asn Asn Pro Pro Gly Ile Asn Arg Asn 
            180                 185                 190         


Ile Tyr Gly Tyr Gln Gly Cys Ala Ala Lys Val Phe Val Pro Ser Lys 
        195                 200                 205             


His Lys Met Val Ser Leu Pro Lys Glu Tyr Glu Gly Tyr Asn Arg Asp 
    210                 215                 220                 


Pro Asn Leu Ser Leu Ala Gly Phe Arg Asn Arg Leu Glu Ile Pro Glu 
225                 230                 235                 240 


Gly Glu Pro Gly His Val Pro Trp Phe Gln Arg Met Asp Ile Pro Glu 
                245                 250                 255     


Gly Gln Ile Gly His Val Asn Lys Ile Gln Arg Phe Asn Phe Val His 
            260                 265                 270         


Gly Lys Asn Ser Gly Lys Val Lys Phe Ser Asp Lys Thr Gly Arg Val 
        275                 280                 285             


Lys Arg Tyr His His Ser Lys Tyr Lys Asp Ala Thr Lys Pro Tyr Lys 
    290                 295                 300                 


Phe Leu Glu Glu Ser Lys Lys Val Ser Ala Leu Asp Ser Ile Leu Ala 
305                 310                 315                 320 


Ile Ile Thr Ile Gly Asp Asp Trp Val Val Phe Asp Ile Arg Gly Leu 
                325                 330                 335     


Tyr Arg Asn Val Phe Tyr Arg Glu Leu Ala Gln Lys Gly Leu Thr Ala 
            340                 345                 350         


Val Gln Leu Leu Asp Leu Phe Thr Gly Asp Pro Val Ile Asp Pro Lys 
        355                 360                 365             


Lys Gly Val Val Thr Phe Ser Tyr Lys Glu Gly Val Val Pro Val Phe 
    370                 375                 380                 


Ser Gln Lys Ile Val Pro Arg Phe Lys Ser Arg Asp Thr Leu Glu Lys 
385                 390                 395                 400 


Leu Thr Ser Gln Gly Pro Val Ala Leu Leu Ser Val Asp Leu Gly Gln 
                405                 410                 415     


Asn Glu Pro Val Ala Ala Arg Val Cys Ser Leu Lys Asn Ile Asn Asp 
            420                 425                 430         


Lys Ile Thr Leu Asp Asn Ser Cys Arg Ile Ser Phe Leu Asp Asp Tyr 
        435                 440                 445             


Lys Lys Gln Ile Lys Asp Tyr Arg Asp Ser Leu Asp Glu Leu Glu Ile 
    450                 455                 460                 


Lys Ile Arg Leu Glu Ala Ile Asn Ser Leu Glu Thr Asn Gln Gln Val 
465                 470                 475                 480 


Glu Ile Arg Asp Leu Asp Val Phe Ser Ala Asp Arg Ala Lys Ala Asn 
                485                 490                 495     


Thr Val Asp Met Phe Asp Ile Asp Pro Asn Leu Ile Ser Trp Asp Ser 
            500                 505                 510         


Met Ser Asp Ala Arg Val Ser Thr Gln Ile Ser Asp Leu Tyr Leu Lys 
        515                 520                 525             


Asn Gly Gly Asp Glu Ser Arg Val Tyr Phe Glu Ile Asn Asn Lys Arg 
    530                 535                 540                 


Ile Lys Arg Ser Asp Tyr Asn Ile Ser Gln Leu Val Arg Pro Lys Leu 
545                 550                 555                 560 


Ser Asp Ser Thr Arg Lys Asn Leu Asn Asp Ser Ile Trp Lys Leu Lys 
                565                 570                 575     


Arg Thr Ser Glu Glu Tyr Leu Lys Leu Ser Lys Arg Lys Leu Glu Leu 
            580                 585                 590         


Ser Arg Ala Val Val Asn Tyr Thr Ile Arg Gln Ser Lys Leu Leu Ser 
        595                 600                 605             


Gly Ile Asn Asp Ile Val Ile Ile Leu Glu Asp Leu Asp Val Lys Lys 
    610                 615                 620                 


Lys Phe Asn Gly Arg Gly Ile Arg Asp Ile Gly Trp Asp Asn Phe Phe 
625                 630                 635                 640 


Ser Ser Arg Lys Glu Asn Arg Trp Phe Ile Pro Ala Phe His Lys Ala 
                645                 650                 655     


Phe Ser Glu Leu Ser Ser Asn Arg Gly Leu Cys Val Ile Glu Val Asn 
            660                 665                 670         


Pro Ala Trp Thr Ser Ala Thr Cys Pro Asp Cys Gly Phe Cys Ser Lys 
        675                 680                 685             


Glu Asn Arg Asp Gly Ile Asn Phe Thr Cys Arg Lys Cys Gly Val Ser 
    690                 695                 700                 


Tyr His Ala Asp Ile Asp Val Ala Thr Leu Asn Ile Ala Arg Val Ala 
705                 710                 715                 720 


Val Leu Gly Lys Pro Met Ser Gly Pro Ala Asp Arg Glu Arg Leu Gly 
                725                 730                 735     


Asp Thr Lys Lys Pro Arg Val Ala Arg Ser Arg Lys Thr Met Lys Arg 
            740                 745                 750         


Lys Asp Ile Ser Asn Ser Thr Val Glu Ala Met Val Thr Ala 
        755                 760                 765     


<210>  361
<211>  757
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasF-2 (nuclease deficient CasF-2)

<400>  361

Met Pro Lys Pro Ala Val Glu Ser Glu Phe Ser Lys Val Leu Lys Lys 
1               5                   10                  15      


His Phe Pro Gly Glu Arg Phe Arg Ser Ser Tyr Met Lys Arg Gly Gly 
            20                  25                  30          


Lys Ile Leu Ala Ala Gln Gly Glu Glu Ala Val Val Ala Tyr Leu Gln 
        35                  40                  45              


Gly Lys Ser Glu Glu Glu Pro Pro Asn Phe Gln Pro Pro Ala Lys Cys 
    50                  55                  60                  


His Val Val Thr Lys Ser Arg Asp Phe Ala Glu Trp Pro Ile Met Lys 
65                  70                  75                  80  


Ala Ser Glu Ala Ile Gln Arg Tyr Ile Tyr Ala Leu Ser Thr Thr Glu 
                85                  90                  95      


Arg Ala Ala Cys Lys Pro Gly Lys Ser Ser Glu Ser His Ala Ala Trp 
            100                 105                 110         


Phe Ala Ala Thr Gly Val Ser Asn His Gly Tyr Ser His Val Gln Gly 
        115                 120                 125             


Leu Asn Leu Ile Phe Asp His Thr Leu Gly Arg Tyr Asp Gly Val Leu 
    130                 135                 140                 


Lys Lys Val Gln Leu Arg Asn Glu Lys Ala Arg Ala Arg Leu Glu Ser 
145                 150                 155                 160 


Ile Asn Ala Ser Arg Ala Asp Glu Gly Leu Pro Glu Ile Lys Ala Glu 
                165                 170                 175     


Glu Glu Glu Val Ala Thr Asn Glu Thr Gly His Leu Leu Gln Pro Pro 
            180                 185                 190         


Gly Ile Asn Pro Ser Phe Tyr Val Tyr Gln Thr Ile Ser Pro Gln Ala 
        195                 200                 205             


Tyr Arg Pro Arg Asp Glu Ile Val Leu Pro Pro Glu Tyr Ala Gly Tyr 
    210                 215                 220                 


Val Arg Asp Pro Asn Ala Pro Ile Pro Leu Gly Val Val Arg Asn Arg 
225                 230                 235                 240 


Cys Asp Ile Gln Lys Gly Cys Pro Gly Tyr Ile Pro Glu Trp Gln Arg 
                245                 250                 255     


Glu Ala Gly Thr Ala Ile Ser Pro Lys Thr Gly Lys Ala Val Thr Val 
            260                 265                 270         


Pro Gly Leu Ser Pro Lys Lys Asn Lys Arg Met Arg Arg Tyr Trp Arg 
        275                 280                 285             


Ser Glu Lys Glu Lys Ala Gln Asp Ala Leu Leu Val Thr Val Arg Ile 
    290                 295                 300                 


Gly Thr Asp Trp Val Val Ile Asp Val Arg Gly Leu Leu Arg Asn Ala 
305                 310                 315                 320 


Arg Trp Arg Thr Ile Ala Pro Lys Asp Ile Ser Leu Asn Ala Leu Leu 
                325                 330                 335     


Asp Leu Phe Thr Gly Asp Pro Val Ile Asp Val Arg Arg Asn Ile Val 
            340                 345                 350         


Thr Phe Thr Tyr Thr Leu Asp Ala Cys Gly Thr Tyr Ala Arg Lys Trp 
        355                 360                 365             


Thr Leu Lys Gly Lys Gln Thr Lys Ala Thr Leu Asp Lys Leu Thr Ala 
    370                 375                 380                 


Thr Gln Thr Val Ala Leu Val Ala Ile Ala Leu Gly Gln Thr Asn Pro 
385                 390                 395                 400 


Ile Ser Ala Gly Ile Ser Arg Val Thr Gln Glu Asn Gly Ala Leu Gln 
                405                 410                 415     


Cys Glu Pro Leu Asp Arg Phe Thr Leu Pro Asp Asp Leu Leu Lys Asp 
            420                 425                 430         


Ile Ser Ala Tyr Arg Ile Ala Trp Asp Arg Asn Glu Glu Glu Leu Arg 
        435                 440                 445             


Ala Arg Ser Val Glu Ala Leu Pro Glu Ala Gln Gln Ala Glu Val Arg 
    450                 455                 460                 


Ala Leu Asp Gly Val Ser Lys Glu Thr Ala Arg Thr Gln Leu Cys Ala 
465                 470                 475                 480 


Asp Phe Gly Leu Asp Pro Lys Arg Leu Pro Trp Asp Lys Met Ser Ser 
                485                 490                 495     


Asn Thr Thr Phe Ile Ser Glu Ala Leu Leu Ser Asn Ser Val Ser Arg 
            500                 505                 510         


Asp Gln Val Phe Phe Thr Pro Ala Pro Lys Lys Gly Ala Lys Lys Lys 
        515                 520                 525             


Ala Pro Val Glu Val Met Arg Lys Asp Arg Thr Trp Ala Arg Ala Tyr 
    530                 535                 540                 


Lys Pro Arg Leu Ser Val Glu Ala Gln Lys Leu Lys Asn Glu Ala Leu 
545                 550                 555                 560 


Trp Ala Leu Lys Arg Thr Ser Pro Glu Tyr Leu Lys Leu Ser Arg Arg 
                565                 570                 575     


Lys Glu Glu Leu Cys Arg Arg Ser Ile Asn Tyr Val Ile Glu Lys Thr 
            580                 585                 590         


Arg Arg Arg Thr Gln Cys Gln Ile Val Ile Pro Val Ile Glu Asp Leu 
        595                 600                 605             


Asn Val Arg Phe Phe His Gly Ser Gly Lys Arg Leu Pro Gly Trp Asp 
    610                 615                 620                 


Asn Phe Phe Thr Ala Lys Lys Glu Asn Arg Trp Phe Ile Gln Gly Leu 
625                 630                 635                 640 


His Lys Ala Phe Ser Asp Leu Arg Thr His Arg Ser Phe Tyr Val Phe 
                645                 650                 655     


Glu Val Arg Pro Glu Arg Thr Ser Ile Thr Cys Pro Lys Cys Gly His 
            660                 665                 670         


Cys Glu Val Gly Asn Arg Asp Gly Glu Ala Phe Gln Cys Leu Ser Cys 
        675                 680                 685             


Gly Lys Thr Cys Asn Ala Asp Leu Asp Val Ala Thr His Asn Leu Thr 
    690                 695                 700                 


Gln Val Ala Leu Thr Gly Lys Thr Met Pro Lys Arg Glu Glu Pro Arg 
705                 710                 715                 720 


Asp Ala Gln Gly Thr Ala Pro Ala Arg Lys Thr Lys Lys Ala Ser Lys 
                725                 730                 735     


Ser Lys Ala Pro Pro Ala Glu Arg Glu Asp Gln Thr Pro Ala Gln Glu 
            740                 745                 750         


Pro Ser Gln Thr Ser 
        755         


<210>  362
<211>  757
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasF-2 RuvCdel1

<400>  362

Met Pro Lys Pro Ala Val Glu Ser Glu Phe Ser Lys Val Leu Lys Lys 
1               5                   10                  15      


His Phe Pro Gly Glu Arg Phe Arg Ser Ser Tyr Met Lys Arg Gly Gly 
            20                  25                  30          


Lys Ile Leu Ala Ala Gln Gly Glu Glu Ala Val Val Ala Tyr Leu Gln 
        35                  40                  45              


Gly Lys Ser Glu Glu Glu Pro Pro Asn Phe Gln Pro Pro Ala Lys Cys 
    50                  55                  60                  


His Val Val Thr Lys Ser Arg Asp Phe Ala Glu Trp Pro Ile Met Lys 
65                  70                  75                  80  


Ala Ser Glu Ala Ile Gln Arg Tyr Ile Tyr Ala Leu Ser Thr Thr Glu 
                85                  90                  95      


Arg Ala Ala Cys Lys Pro Gly Lys Ser Ser Glu Ser His Ala Ala Trp 
            100                 105                 110         


Phe Ala Ala Thr Gly Val Ser Asn His Gly Tyr Ser His Val Gln Gly 
        115                 120                 125             


Leu Asn Leu Ile Phe Asp His Thr Leu Gly Arg Tyr Asp Gly Val Leu 
    130                 135                 140                 


Lys Lys Val Gln Leu Arg Asn Glu Lys Ala Arg Ala Arg Leu Glu Ser 
145                 150                 155                 160 


Ile Asn Ala Ser Arg Ala Asp Glu Gly Leu Pro Glu Ile Lys Ala Glu 
                165                 170                 175     


Glu Glu Glu Val Ala Thr Asn Glu Thr Gly His Leu Leu Gln Pro Pro 
            180                 185                 190         


Gly Ile Asn Pro Ser Phe Tyr Val Tyr Gln Thr Ile Ser Pro Gln Ala 
        195                 200                 205             


Tyr Arg Pro Arg Asp Glu Ile Val Leu Pro Pro Glu Tyr Ala Gly Tyr 
    210                 215                 220                 


Val Arg Asp Pro Asn Ala Pro Ile Pro Leu Gly Val Val Arg Asn Arg 
225                 230                 235                 240 


Cys Asp Ile Gln Lys Gly Cys Pro Gly Tyr Ile Pro Glu Trp Gln Arg 
                245                 250                 255     


Glu Ala Gly Thr Ala Ile Ser Pro Lys Thr Gly Lys Ala Val Thr Val 
            260                 265                 270         


Pro Gly Leu Ser Pro Lys Lys Asn Lys Arg Met Arg Arg Tyr Trp Arg 
        275                 280                 285             


Ser Glu Lys Glu Lys Ala Gln Asp Ala Leu Leu Val Thr Val Arg Ile 
    290                 295                 300                 


Gly Thr Asp Trp Val Val Ile Asp Val Arg Gly Leu Leu Arg Asn Ala 
305                 310                 315                 320 


Arg Trp Arg Thr Ile Ala Pro Lys Asp Ile Ser Leu Asn Ala Leu Leu 
                325                 330                 335     


Asp Leu Phe Thr Gly Asp Pro Val Ile Asp Val Arg Arg Asn Ile Val 
            340                 345                 350         


Thr Phe Thr Tyr Thr Leu Asp Ala Cys Gly Thr Tyr Ala Arg Lys Trp 
        355                 360                 365             


Thr Leu Lys Gly Lys Gln Thr Lys Ala Thr Leu Asp Lys Leu Thr Ala 
    370                 375                 380                 


Thr Gln Thr Val Ala Leu Val Ala Ile Ala Leu Gly Gln Thr Asn Pro 
385                 390                 395                 400 


Ile Ser Ala Gly Ile Ser Arg Val Thr Gln Glu Asn Gly Ala Leu Gln 
                405                 410                 415     


Cys Glu Pro Leu Asp Arg Phe Thr Leu Pro Asp Asp Leu Leu Lys Asp 
            420                 425                 430         


Ile Ser Ala Tyr Arg Ile Ala Trp Asp Arg Asn Glu Glu Glu Leu Arg 
        435                 440                 445             


Ala Arg Ser Val Glu Ala Leu Pro Glu Ala Gln Gln Ala Glu Val Arg 
    450                 455                 460                 


Ala Leu Asp Gly Val Ser Lys Glu Thr Ala Arg Thr Gln Leu Cys Ala 
465                 470                 475                 480 


Asp Phe Gly Leu Asp Pro Lys Arg Leu Pro Trp Asp Lys Met Ser Ser 
                485                 490                 495     


Asn Thr Thr Phe Ile Ser Glu Ala Leu Leu Ser Asn Ser Val Ser Arg 
            500                 505                 510         


Asp Gln Val Phe Phe Thr Pro Ala Pro Lys Lys Gly Ala Lys Lys Lys 
        515                 520                 525             


Ala Pro Val Glu Val Met Arg Lys Asp Arg Thr Trp Ala Arg Ala Tyr 
    530                 535                 540                 


Lys Pro Arg Leu Ser Val Glu Ala Gln Lys Leu Lys Asn Glu Ala Leu 
545                 550                 555                 560 


Trp Ala Leu Lys Arg Thr Ser Pro Glu Tyr Leu Lys Leu Ser Arg Arg 
                565                 570                 575     


Lys Glu Glu Leu Cys Arg Arg Ser Ile Asn Tyr Val Ile Glu Lys Thr 
            580                 585                 590         


Arg Arg Arg Thr Gln Cys Gln Ile Val Ile Pro Val Ile Glu Asp Leu 
        595                 600                 605             


Asn Val Arg Phe Phe His Gly Ser Gly Lys Arg Leu Pro Gly Trp Asp 
    610                 615                 620                 


Asn Phe Phe Thr Ala Lys Lys Glu Asn Arg Trp Phe Ile Gln Gly Leu 
625                 630                 635                 640 


His Lys Ala Phe Ser Asp Leu Arg Thr His Arg Ser Phe Tyr Val Phe 
                645                 650                 655     


Glu Val Arg Pro Glu Arg Thr Ser Ile Thr Cys Pro Lys Cys Gly His 
            660                 665                 670         


Cys Glu Val Gly Asn Arg Asp Gly Glu Ala Phe Gln Cys Leu Ser Cys 
        675                 680                 685             


Gly Lys Thr Cys Asn Ala Asp Leu Asp Val Ala Thr His Asn Leu Thr 
    690                 695                 700                 


Gln Val Ala Leu Thr Gly Lys Thr Met Pro Lys Arg Glu Glu Pro Arg 
705                 710                 715                 720 


Asp Ala Gln Gly Thr Ala Pro Ala Arg Lys Thr Lys Lys Ala Ser Lys 
                725                 730                 735     


Ser Lys Ala Pro Pro Ala Glu Arg Glu Asp Gln Thr Pro Ala Gln Glu 
            740                 745                 750         


Pro Ser Gln Thr Ser 
        755         


<210>  363
<211>  983
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasF2-Fok (N-terminal Fok)

<400>  363

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Pro Lys Pro Ala Val Glu Ser Glu Phe Ser Lys Val Leu 
225                 230                 235                 240 


Lys Lys His Phe Pro Gly Glu Arg Phe Arg Ser Ser Tyr Met Lys Arg 
                245                 250                 255     


Gly Gly Lys Ile Leu Ala Ala Gln Gly Glu Glu Ala Val Val Ala Tyr 
            260                 265                 270         


Leu Gln Gly Lys Ser Glu Glu Glu Pro Pro Asn Phe Gln Pro Pro Ala 
        275                 280                 285             


Lys Cys His Val Val Thr Lys Ser Arg Asp Phe Ala Glu Trp Pro Ile 
    290                 295                 300                 


Met Lys Ala Ser Glu Ala Ile Gln Arg Tyr Ile Tyr Ala Leu Ser Thr 
305                 310                 315                 320 


Thr Glu Arg Ala Ala Cys Lys Pro Gly Lys Ser Ser Glu Ser His Ala 
                325                 330                 335     


Ala Trp Phe Ala Ala Thr Gly Val Ser Asn His Gly Tyr Ser His Val 
            340                 345                 350         


Gln Gly Leu Asn Leu Ile Phe Asp His Thr Leu Gly Arg Tyr Asp Gly 
        355                 360                 365             


Val Leu Lys Lys Val Gln Leu Arg Asn Glu Lys Ala Arg Ala Arg Leu 
    370                 375                 380                 


Glu Ser Ile Asn Ala Ser Arg Ala Asp Glu Gly Leu Pro Glu Ile Lys 
385                 390                 395                 400 


Ala Glu Glu Glu Glu Val Ala Thr Asn Glu Thr Gly His Leu Leu Gln 
                405                 410                 415     


Pro Pro Gly Ile Asn Pro Ser Phe Tyr Val Tyr Gln Thr Ile Ser Pro 
            420                 425                 430         


Gln Ala Tyr Arg Pro Arg Asp Glu Ile Val Leu Pro Pro Glu Tyr Ala 
        435                 440                 445             


Gly Tyr Val Arg Asp Pro Asn Ala Pro Ile Pro Leu Gly Val Val Arg 
    450                 455                 460                 


Asn Arg Cys Asp Ile Gln Lys Gly Cys Pro Gly Tyr Ile Pro Glu Trp 
465                 470                 475                 480 


Gln Arg Glu Ala Gly Thr Ala Ile Ser Pro Lys Thr Gly Lys Ala Val 
                485                 490                 495     


Thr Val Pro Gly Leu Ser Pro Lys Lys Asn Lys Arg Met Arg Arg Tyr 
            500                 505                 510         


Trp Arg Ser Glu Lys Glu Lys Ala Gln Asp Ala Leu Leu Val Thr Val 
        515                 520                 525             


Arg Ile Gly Thr Asp Trp Val Val Ile Asp Val Arg Gly Leu Leu Arg 
    530                 535                 540                 


Asn Ala Arg Trp Arg Thr Ile Ala Pro Lys Asp Ile Ser Leu Asn Ala 
545                 550                 555                 560 


Leu Leu Asp Leu Phe Thr Gly Asp Pro Val Ile Asp Val Arg Arg Asn 
                565                 570                 575     


Ile Val Thr Phe Thr Tyr Thr Leu Asp Ala Cys Gly Thr Tyr Ala Arg 
            580                 585                 590         


Lys Trp Thr Leu Lys Gly Lys Gln Thr Lys Ala Thr Leu Asp Lys Leu 
        595                 600                 605             


Thr Ala Thr Gln Thr Val Ala Leu Val Ala Ile Ala Leu Gly Gln Thr 
    610                 615                 620                 


Asn Pro Ile Ser Ala Gly Ile Ser Arg Val Thr Gln Glu Asn Gly Ala 
625                 630                 635                 640 


Leu Gln Cys Glu Pro Leu Asp Arg Phe Thr Leu Pro Asp Asp Leu Leu 
                645                 650                 655     


Lys Asp Ile Ser Ala Tyr Arg Ile Ala Trp Asp Arg Asn Glu Glu Glu 
            660                 665                 670         


Leu Arg Ala Arg Ser Val Glu Ala Leu Pro Glu Ala Gln Gln Ala Glu 
        675                 680                 685             


Val Arg Ala Leu Asp Gly Val Ser Lys Glu Thr Ala Arg Thr Gln Leu 
    690                 695                 700                 


Cys Ala Asp Phe Gly Leu Asp Pro Lys Arg Leu Pro Trp Asp Lys Met 
705                 710                 715                 720 


Ser Ser Asn Thr Thr Phe Ile Ser Glu Ala Leu Leu Ser Asn Ser Val 
                725                 730                 735     


Ser Arg Asp Gln Val Phe Phe Thr Pro Ala Pro Lys Lys Gly Ala Lys 
            740                 745                 750         


Lys Lys Ala Pro Val Glu Val Met Arg Lys Asp Arg Thr Trp Ala Arg 
        755                 760                 765             


Ala Tyr Lys Pro Arg Leu Ser Val Glu Ala Gln Lys Leu Lys Asn Glu 
    770                 775                 780                 


Ala Leu Trp Ala Leu Lys Arg Thr Ser Pro Glu Tyr Leu Lys Leu Ser 
785                 790                 795                 800 


Arg Arg Lys Glu Glu Leu Cys Arg Arg Ser Ile Asn Tyr Val Ile Glu 
                805                 810                 815     


Lys Thr Arg Arg Arg Thr Gln Cys Gln Ile Val Ile Pro Val Ile Glu 
            820                 825                 830         


Asp Leu Asn Val Arg Phe Phe His Gly Ser Gly Lys Arg Leu Pro Gly 
        835                 840                 845             


Trp Asp Asn Phe Phe Thr Ala Lys Lys Glu Asn Arg Trp Phe Ile Gln 
    850                 855                 860                 


Gly Leu His Lys Ala Phe Ser Asp Leu Arg Thr His Arg Ser Phe Tyr 
865                 870                 875                 880 


Val Phe Glu Val Arg Pro Glu Arg Thr Ser Ile Thr Cys Pro Lys Cys 
                885                 890                 895     


Gly His Cys Glu Val Gly Asn Arg Asp Gly Glu Ala Phe Gln Cys Leu 
            900                 905                 910         


Ser Cys Gly Lys Thr Cys Asn Ala Asp Leu Asp Val Ala Thr His Asn 
        915                 920                 925             


Leu Thr Gln Val Ala Leu Thr Gly Lys Thr Met Pro Lys Arg Glu Glu 
    930                 935                 940                 


Pro Arg Asp Ala Gln Gly Thr Ala Pro Ala Arg Lys Thr Lys Lys Ala 
945                 950                 955                 960 


Ser Lys Ser Lys Ala Pro Pro Ala Glu Arg Glu Asp Gln Thr Pro Ala 
                965                 970                 975     


Gln Glu Pro Ser Gln Thr Ser 
            980             


<210>  364
<211>  975
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasF2-Fok RuvC del 2

<400>  364

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Pro Lys Pro Ala Val Glu Ser Glu Phe Ser Lys Val Leu 
225                 230                 235                 240 


Lys Lys His Phe Pro Gly Glu Arg Phe Arg Ser Ser Tyr Met Lys Arg 
                245                 250                 255     


Gly Gly Lys Ile Leu Ala Ala Gln Gly Glu Glu Ala Val Val Ala Tyr 
            260                 265                 270         


Leu Gln Gly Lys Ser Glu Glu Glu Pro Pro Asn Phe Gln Pro Pro Ala 
        275                 280                 285             


Lys Cys His Val Val Thr Lys Ser Arg Asp Phe Ala Glu Trp Pro Ile 
    290                 295                 300                 


Met Lys Ala Ser Glu Ala Ile Gln Arg Tyr Ile Tyr Ala Leu Ser Thr 
305                 310                 315                 320 


Thr Glu Arg Ala Ala Cys Lys Pro Gly Lys Ser Ser Glu Ser His Ala 
                325                 330                 335     


Ala Trp Phe Ala Ala Thr Gly Val Ser Asn His Gly Tyr Ser His Val 
            340                 345                 350         


Gln Gly Leu Asn Leu Ile Phe Asp His Thr Leu Gly Arg Tyr Asp Gly 
        355                 360                 365             


Val Leu Lys Lys Val Gln Leu Arg Asn Glu Lys Ala Arg Ala Arg Leu 
    370                 375                 380                 


Glu Ser Ile Asn Ala Ser Arg Ala Asp Glu Gly Leu Pro Glu Ile Lys 
385                 390                 395                 400 


Ala Glu Glu Glu Glu Val Ala Thr Asn Glu Thr Gly His Leu Leu Gln 
                405                 410                 415     


Pro Pro Gly Ile Asn Pro Ser Phe Tyr Val Tyr Gln Thr Ile Ser Pro 
            420                 425                 430         


Gln Ala Tyr Arg Pro Arg Asp Glu Ile Val Leu Pro Pro Glu Tyr Ala 
        435                 440                 445             


Gly Tyr Val Arg Asp Pro Asn Ala Pro Ile Pro Leu Gly Val Val Arg 
    450                 455                 460                 


Asn Arg Cys Asp Ile Gln Lys Gly Cys Pro Gly Tyr Ile Pro Glu Trp 
465                 470                 475                 480 


Gln Arg Glu Ala Gly Thr Ala Ile Ser Pro Lys Thr Gly Lys Ala Val 
                485                 490                 495     


Thr Val Pro Gly Leu Ser Pro Lys Lys Asn Lys Arg Met Arg Arg Tyr 
            500                 505                 510         


Trp Arg Ser Glu Lys Glu Lys Ala Gln Asp Ala Leu Leu Val Thr Val 
        515                 520                 525             


Arg Ile Gly Thr Asp Trp Val Val Ile Asp Val Arg Gly Leu Leu Arg 
    530                 535                 540                 


Asn Ala Arg Trp Arg Thr Ile Ala Pro Lys Asp Ile Ser Leu Asn Ala 
545                 550                 555                 560 


Leu Leu Asp Leu Phe Thr Gly Asp Pro Val Ile Asp Val Arg Arg Asn 
                565                 570                 575     


Ile Val Thr Phe Thr Tyr Thr Leu Asp Ala Cys Gly Thr Tyr Ala Arg 
            580                 585                 590         


Lys Trp Thr Leu Lys Gly Lys Gln Thr Lys Ala Thr Leu Asp Lys Leu 
        595                 600                 605             


Thr Ala Thr Gln Thr Val Ala Leu Val Ala Ile Ala Leu Gly Gln Thr 
    610                 615                 620                 


Asn Pro Ile Ser Ala Gly Ile Ser Arg Val Thr Gln Glu Asn Gly Ala 
625                 630                 635                 640 


Leu Gln Cys Glu Pro Leu Asp Arg Phe Thr Leu Pro Asp Asp Leu Leu 
                645                 650                 655     


Lys Asp Ile Ser Ala Tyr Arg Ile Ala Trp Asp Arg Asn Glu Glu Glu 
            660                 665                 670         


Leu Arg Ala Arg Ser Val Glu Ala Leu Pro Glu Ala Gln Gln Ala Glu 
        675                 680                 685             


Val Arg Ala Leu Asp Gly Val Ser Lys Glu Thr Ala Arg Thr Gln Leu 
    690                 695                 700                 


Cys Ala Asp Phe Gly Leu Asp Pro Lys Arg Leu Pro Trp Asp Lys Met 
705                 710                 715                 720 


Ser Ser Asn Thr Thr Phe Ile Ser Glu Ala Leu Leu Ser Asn Ser Val 
                725                 730                 735     


Ser Arg Asp Gln Val Phe Phe Thr Pro Ala Pro Lys Lys Gly Ala Lys 
            740                 745                 750         


Lys Lys Ala Pro Val Glu Val Met Arg Lys Asp Arg Thr Trp Ala Arg 
        755                 760                 765             


Ala Tyr Lys Pro Arg Leu Ser Val Glu Ala Gln Lys Leu Lys Asn Glu 
    770                 775                 780                 


Ala Leu Trp Ala Leu Lys Arg Thr Ser Pro Glu Tyr Leu Lys Leu Ser 
785                 790                 795                 800 


Arg Arg Lys Glu Glu Leu Cys Arg Arg Ser Ile Asn Tyr Val Ile Glu 
                805                 810                 815     


Lys Thr Arg Arg Arg Thr Gln Cys Gln Ile Gly Gly Ser Gly Phe His 
            820                 825                 830         


Gly Ser Gly Lys Arg Leu Pro Gly Trp Asp Asn Phe Phe Thr Ala Lys 
        835                 840                 845             


Lys Glu Asn Arg Trp Phe Ile Gln Gly Leu His Lys Ala Phe Ser Asp 
    850                 855                 860                 


Leu Arg Thr His Arg Ser Phe Tyr Val Phe Glu Val Arg Pro Glu Arg 
865                 870                 875                 880 


Thr Ser Ile Thr Cys Pro Lys Cys Gly His Cys Glu Val Gly Asn Arg 
                885                 890                 895     


Asp Gly Glu Ala Phe Gln Cys Leu Ser Cys Gly Lys Thr Cys Asn Ala 
            900                 905                 910         


Asp Leu Asp Val Ala Thr His Asn Leu Thr Gln Val Ala Leu Thr Gly 
        915                 920                 925             


Lys Thr Met Pro Lys Arg Glu Glu Pro Arg Asp Ala Gln Gly Thr Ala 
    930                 935                 940                 


Pro Ala Arg Lys Thr Lys Lys Ala Ser Lys Ser Lys Ala Pro Pro Ala 
945                 950                 955                 960 


Glu Arg Glu Asp Gln Thr Pro Ala Gln Glu Pro Ser Gln Thr Ser 
                965                 970                 975 


<210>  365
<211>  969
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasF2-Fok RuvC del 3

<400>  365

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser Gln Leu 
1               5                   10                  15      


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
            20                  25                  30          


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
        35                  40                  45              


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
    50                  55                  60                  


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
65                  70                  75                  80  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
                85                  90                  95      


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
            100                 105                 110         


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
        115                 120                 125             


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
    130                 135                 140                 


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
145                 150                 155                 160 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
                165                 170                 175     


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
            180                 185                 190         


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
        195                 200                 205             


Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro 
    210                 215                 220                 


Glu Ser Met Pro Lys Pro Ala Val Glu Ser Glu Phe Ser Lys Val Leu 
225                 230                 235                 240 


Lys Lys His Phe Pro Gly Glu Arg Phe Arg Ser Ser Tyr Met Lys Arg 
                245                 250                 255     


Gly Gly Lys Ile Leu Ala Ala Gln Gly Glu Glu Ala Val Val Ala Tyr 
            260                 265                 270         


Leu Gln Gly Lys Ser Glu Glu Glu Pro Pro Asn Phe Gln Pro Pro Ala 
        275                 280                 285             


Lys Cys His Val Val Thr Lys Ser Arg Asp Phe Ala Glu Trp Pro Ile 
    290                 295                 300                 


Met Lys Ala Ser Glu Ala Ile Gln Arg Tyr Ile Tyr Ala Leu Ser Thr 
305                 310                 315                 320 


Thr Glu Arg Ala Ala Cys Lys Pro Gly Lys Ser Ser Glu Ser His Ala 
                325                 330                 335     


Ala Trp Phe Ala Ala Thr Gly Val Ser Asn His Gly Tyr Ser His Val 
            340                 345                 350         


Gln Gly Leu Asn Leu Ile Phe Asp His Thr Leu Gly Arg Tyr Asp Gly 
        355                 360                 365             


Val Leu Lys Lys Val Gln Leu Arg Asn Glu Lys Ala Arg Ala Arg Leu 
    370                 375                 380                 


Glu Ser Ile Asn Ala Ser Arg Ala Asp Glu Gly Leu Pro Glu Ile Lys 
385                 390                 395                 400 


Ala Glu Glu Glu Glu Val Ala Thr Asn Glu Thr Gly His Leu Leu Gln 
                405                 410                 415     


Pro Pro Gly Ile Asn Pro Ser Phe Tyr Val Tyr Gln Thr Ile Ser Pro 
            420                 425                 430         


Gln Ala Tyr Arg Pro Arg Asp Glu Ile Val Leu Pro Pro Glu Tyr Ala 
        435                 440                 445             


Gly Tyr Val Arg Asp Pro Asn Ala Pro Ile Pro Leu Gly Val Val Arg 
    450                 455                 460                 


Asn Arg Cys Asp Ile Gln Lys Gly Cys Pro Gly Tyr Ile Pro Glu Trp 
465                 470                 475                 480 


Gln Arg Glu Ala Gly Thr Ala Ile Ser Pro Lys Thr Gly Lys Ala Val 
                485                 490                 495     


Thr Val Pro Gly Leu Ser Pro Lys Lys Asn Lys Arg Met Arg Arg Tyr 
            500                 505                 510         


Trp Arg Ser Glu Lys Glu Lys Ala Gln Asp Ala Leu Leu Val Thr Val 
        515                 520                 525             


Arg Ile Gly Thr Asp Trp Val Val Ile Asp Val Arg Gly Leu Leu Arg 
    530                 535                 540                 


Asn Ala Arg Trp Arg Thr Ile Ala Pro Lys Asp Ile Ser Leu Asn Ala 
545                 550                 555                 560 


Leu Leu Asp Leu Phe Thr Gly Asp Pro Val Ile Asp Val Arg Arg Asn 
                565                 570                 575     


Ile Val Thr Phe Thr Tyr Thr Leu Asp Ala Cys Gly Thr Tyr Ala Arg 
            580                 585                 590         


Lys Trp Thr Leu Lys Gly Lys Gln Thr Lys Ala Thr Leu Asp Lys Leu 
        595                 600                 605             


Thr Ala Thr Gln Thr Val Ala Leu Val Ala Ile Ala Leu Gly Gln Thr 
    610                 615                 620                 


Asn Pro Ile Ser Ala Gly Ile Ser Arg Val Thr Gln Glu Asn Gly Ala 
625                 630                 635                 640 


Leu Gln Cys Glu Pro Leu Asp Arg Phe Thr Leu Pro Asp Asp Leu Leu 
                645                 650                 655     


Lys Asp Ile Ser Ala Tyr Arg Ile Ala Trp Asp Arg Asn Glu Glu Glu 
            660                 665                 670         


Leu Arg Ala Arg Ser Val Glu Ala Leu Pro Glu Ala Gln Gln Ala Glu 
        675                 680                 685             


Val Arg Ala Leu Asp Gly Val Ser Lys Glu Thr Ala Arg Thr Gln Leu 
    690                 695                 700                 


Cys Ala Asp Phe Gly Leu Asp Pro Lys Arg Leu Pro Trp Asp Lys Met 
705                 710                 715                 720 


Ser Ser Asn Thr Thr Phe Ile Ser Glu Ala Leu Leu Ser Asn Ser Val 
                725                 730                 735     


Ser Arg Asp Gln Val Phe Phe Thr Pro Ala Pro Lys Lys Gly Ala Lys 
            740                 745                 750         


Lys Lys Ala Pro Val Glu Val Met Arg Lys Asp Arg Thr Trp Ala Arg 
        755                 760                 765             


Ala Tyr Lys Pro Arg Leu Ser Val Glu Ala Gln Lys Leu Lys Asn Glu 
    770                 775                 780                 


Ala Leu Trp Ala Leu Lys Arg Thr Ser Pro Glu Tyr Leu Lys Leu Ser 
785                 790                 795                 800 


Arg Arg Lys Glu Glu Leu Cys Arg Arg Ser Ile Asn Tyr Val Ile Glu 
                805                 810                 815     


Lys Thr Arg Arg Arg Thr Gln Cys Gln Ile Val Ile Pro Val Ile Glu 
            820                 825                 830         


Asp Leu Asn Val Arg Phe Phe His Gly Ser Gly Lys Arg Leu Pro Gly 
        835                 840                 845             


Trp Asp Asn Phe Phe Thr Ala Lys Lys Glu Asn Arg Trp Phe Ile Gln 
    850                 855                 860                 


Gly Leu His Lys Ala Phe Ser Asp Leu Arg Thr His Arg Ser Phe Tyr 
865                 870                 875                 880 


Val Phe Glu Val Arg Pro Glu Arg Thr Ser Ile Thr Cys Pro Lys Cys 
                885                 890                 895     


Gly His Cys Glu Val Gly Asn Arg Asp Gly Glu Ala Phe Gln Cys Leu 
            900                 905                 910         


Ser Cys Gly Gly Ser Gly Ala Leu Thr Gly Lys Thr Met Pro Lys Arg 
        915                 920                 925             


Glu Glu Pro Arg Asp Ala Gln Gly Thr Ala Pro Ala Arg Lys Thr Lys 
    930                 935                 940                 


Lys Ala Ser Lys Ser Lys Ala Pro Pro Ala Glu Arg Glu Asp Gln Thr 
945                 950                 955                 960 


Pro Ala Gln Glu Pro Ser Gln Thr Ser 
                965                 


<210>  366
<211>  978
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasF2-Fok (C-terminal Fok)

<400>  366

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Pro Lys Pro 
1               5                   10                  15      


Ala Val Glu Ser Glu Phe Ser Lys Val Leu Lys Lys His Phe Pro Gly 
            20                  25                  30          


Glu Arg Phe Arg Ser Ser Tyr Met Lys Arg Gly Gly Lys Ile Leu Ala 
        35                  40                  45              


Ala Gln Gly Glu Glu Ala Val Val Ala Tyr Leu Gln Gly Lys Ser Glu 
    50                  55                  60                  


Glu Glu Pro Pro Asn Phe Gln Pro Pro Ala Lys Cys His Val Val Thr 
65                  70                  75                  80  


Lys Ser Arg Asp Phe Ala Glu Trp Pro Ile Met Lys Ala Ser Glu Ala 
                85                  90                  95      


Ile Gln Arg Tyr Ile Tyr Ala Leu Ser Thr Thr Glu Arg Ala Ala Cys 
            100                 105                 110         


Lys Pro Gly Lys Ser Ser Glu Ser His Ala Ala Trp Phe Ala Ala Thr 
        115                 120                 125             


Gly Val Ser Asn His Gly Tyr Ser His Val Gln Gly Leu Asn Leu Ile 
    130                 135                 140                 


Phe Asp His Thr Leu Gly Arg Tyr Asp Gly Val Leu Lys Lys Val Gln 
145                 150                 155                 160 


Leu Arg Asn Glu Lys Ala Arg Ala Arg Leu Glu Ser Ile Asn Ala Ser 
                165                 170                 175     


Arg Ala Asp Glu Gly Leu Pro Glu Ile Lys Ala Glu Glu Glu Glu Val 
            180                 185                 190         


Ala Thr Asn Glu Thr Gly His Leu Leu Gln Pro Pro Gly Ile Asn Pro 
        195                 200                 205             


Ser Phe Tyr Val Tyr Gln Thr Ile Ser Pro Gln Ala Tyr Arg Pro Arg 
    210                 215                 220                 


Asp Glu Ile Val Leu Pro Pro Glu Tyr Ala Gly Tyr Val Arg Asp Pro 
225                 230                 235                 240 


Asn Ala Pro Ile Pro Leu Gly Val Val Arg Asn Arg Cys Asp Ile Gln 
                245                 250                 255     


Lys Gly Cys Pro Gly Tyr Ile Pro Glu Trp Gln Arg Glu Ala Gly Thr 
            260                 265                 270         


Ala Ile Ser Pro Lys Thr Gly Lys Ala Val Thr Val Pro Gly Leu Ser 
        275                 280                 285             


Pro Lys Lys Asn Lys Arg Met Arg Arg Tyr Trp Arg Ser Glu Lys Glu 
    290                 295                 300                 


Lys Ala Gln Asp Ala Leu Leu Val Thr Val Arg Ile Gly Thr Asp Trp 
305                 310                 315                 320 


Val Val Ile Asp Val Arg Gly Leu Leu Arg Asn Ala Arg Trp Arg Thr 
                325                 330                 335     


Ile Ala Pro Lys Asp Ile Ser Leu Asn Ala Leu Leu Asp Leu Phe Thr 
            340                 345                 350         


Gly Asp Pro Val Ile Asp Val Arg Arg Asn Ile Val Thr Phe Thr Tyr 
        355                 360                 365             


Thr Leu Asp Ala Cys Gly Thr Tyr Ala Arg Lys Trp Thr Leu Lys Gly 
    370                 375                 380                 


Lys Gln Thr Lys Ala Thr Leu Asp Lys Leu Thr Ala Thr Gln Thr Val 
385                 390                 395                 400 


Ala Leu Val Ala Ile Ala Leu Gly Gln Thr Asn Pro Ile Ser Ala Gly 
                405                 410                 415     


Ile Ser Arg Val Thr Gln Glu Asn Gly Ala Leu Gln Cys Glu Pro Leu 
            420                 425                 430         


Asp Arg Phe Thr Leu Pro Asp Asp Leu Leu Lys Asp Ile Ser Ala Tyr 
        435                 440                 445             


Arg Ile Ala Trp Asp Arg Asn Glu Glu Glu Leu Arg Ala Arg Ser Val 
    450                 455                 460                 


Glu Ala Leu Pro Glu Ala Gln Gln Ala Glu Val Arg Ala Leu Asp Gly 
465                 470                 475                 480 


Val Ser Lys Glu Thr Ala Arg Thr Gln Leu Cys Ala Asp Phe Gly Leu 
                485                 490                 495     


Asp Pro Lys Arg Leu Pro Trp Asp Lys Met Ser Ser Asn Thr Thr Phe 
            500                 505                 510         


Ile Ser Glu Ala Leu Leu Ser Asn Ser Val Ser Arg Asp Gln Val Phe 
        515                 520                 525             


Phe Thr Pro Ala Pro Lys Lys Gly Ala Lys Lys Lys Ala Pro Val Glu 
    530                 535                 540                 


Val Met Arg Lys Asp Arg Thr Trp Ala Arg Ala Tyr Lys Pro Arg Leu 
545                 550                 555                 560 


Ser Val Glu Ala Gln Lys Leu Lys Asn Glu Ala Leu Trp Ala Leu Lys 
                565                 570                 575     


Arg Thr Ser Pro Glu Tyr Leu Lys Leu Ser Arg Arg Lys Glu Glu Leu 
            580                 585                 590         


Cys Arg Arg Ser Ile Asn Tyr Val Ile Glu Lys Thr Arg Arg Arg Thr 
        595                 600                 605             


Gln Cys Gln Ile Val Ile Pro Val Ile Glu Asp Leu Asn Val Arg Phe 
    610                 615                 620                 


Phe His Gly Ser Gly Lys Arg Leu Pro Gly Trp Asp Asn Phe Phe Thr 
625                 630                 635                 640 


Ala Lys Lys Glu Asn Arg Trp Phe Ile Gln Gly Leu His Lys Ala Phe 
                645                 650                 655     


Ser Asp Leu Arg Thr His Arg Ser Phe Tyr Val Phe Glu Val Arg Pro 
            660                 665                 670         


Glu Arg Thr Ser Ile Thr Cys Pro Lys Cys Gly His Cys Glu Val Gly 
        675                 680                 685             


Asn Arg Asp Gly Glu Ala Phe Gln Cys Leu Ser Cys Gly Lys Thr Cys 
    690                 695                 700                 


Asn Ala Asp Leu Asp Val Ala Thr His Asn Leu Thr Gln Val Ala Leu 
705                 710                 715                 720 


Thr Gly Lys Thr Met Pro Lys Arg Glu Glu Pro Arg Asp Ala Gln Gly 
                725                 730                 735     


Thr Ala Pro Ala Arg Lys Thr Lys Lys Ala Ser Lys Ser Lys Ala Pro 
            740                 745                 750         


Pro Ala Glu Arg Glu Asp Gln Thr Pro Ala Gln Glu Pro Ser Gln Thr 
        755                 760                 765             


Ser Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu 
    770                 775                 780                 


Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
785                 790                 795                 800 


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
                805                 810                 815     


Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
            820                 825                 830         


Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro 
        835                 840                 845             


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
    850                 855                 860                 


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
865                 870                 875                 880 


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
                885                 890                 895     


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
            900                 905                 910         


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
        915                 920                 925             


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
    930                 935                 940                 


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
945                 950                 955                 960 


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
                965                 970                 975     


Asn Phe 
        


<210>  367
<211>  593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD1.dcasffok.pamout.1

<400>  367
acgcgtaaaa aaaaagtcac cacgagcagg gctgggtccc ctcgtgaggg gcaatcgttg       60

ggtgtttcgt ggtctcatac agaacttata agattcccaa atccaaagac atttcacgtt      120

tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagggcgcc actcccctgt      180

ccctcacagc catcttcctg ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac      240

tgggcccgcg attccttgga gcgggttgat gacgtcagcg ttcaaagagg gcctatttcc      300

catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa      360

tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct      420

tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac      480

ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacccaacg      540

attgcccctc acgaggggac agaagctgca ggtgaaggtg tttttttacg cgt             593


<210>  368
<211>  593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD1.dcasffok.pamout.2

<400>  368
acgcgtaaaa aaaaagtccc cttcggtcac cacgagtccc ctcgtgaggg gcaatcgttg       60

ggtgtttcgt ggtctcatac agaacttata agattcccaa atccaaagac atttcacgtt      120

tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagggcgcc actcccctgt      180

ccctcacagc catcttcctg ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac      240

tgggcccgcg attccttgga gcgggttgat gacgtcagcg ttcaaagagg gcctatttcc      300

catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa      360

tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct      420

tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac      480

ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacccaacg      540

attgcccctc acgaggggac gatgtgttgg agaagctgca tttttttacg cgt             593


<210>  369
<211>  593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD1.dcasffok.pamout.3

<400>  369
acgcgtaaaa aaaaactggt accgcatgag ccccagtccc ctcgtgaggg gcaatcgttg       60

ggtgtttcgt ggtctcatac agaacttata agattcccaa atccaaagac atttcacgtt      120

tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagggcgcc actcccctgt      180

ccctcacagc catcttcctg ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac      240

tgggcccgcg attccttgga gcgggttgat gacgtcagcg ttcaaagagg gcctatttcc      300

catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa      360

tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct      420

tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac      480

ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacccaacg      540

attgcccctc acgaggggac agcttctcca acacatcgga tttttttacg cgt             593


<210>  370
<211>  593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD1.dcasffok.pamin.1

<400>  370
acgcgtaaaa aaaaactccc cagccctgct cgtgggtccc ctcgtgaggg gcaatcgttg       60

ggtgtttcgt ggtctcatac agaacttata agattcccaa atccaaagac atttcacgtt      120

tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagggcgcc actcccctgt      180

ccctcacagc catcttcctg ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac      240

tgggcccgcg attccttgga gcgggttgat gacgtcagcg ttcaaagagg gcctatttcc      300

catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa      360

tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct      420

tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac      480

ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacccaacg      540

attgcccctc acgaggggac acctgcagct tctccaacac tttttttacg cgt             593


<210>  371
<211>  593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD1.dcasffok.pamin.2

<400>  371
acgcgtaaaa aaaaacccct tcggtcacca cgagcgtccc ctcgtgaggg gcaatcgttg       60

ggtgtttcgt ggtctcatac agaacttata agattcccaa atccaaagac atttcacgtt      120

tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagggcgcc actcccctgt      180

ccctcacagc catcttcctg ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac      240

tgggcccgcg attccttgga gcgggttgat gacgtcagcg ttcaaagagg gcctatttcc      300

catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa      360

tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct      420

tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac      480

ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacccaacg      540

attgcccctc acgaggggac tccaacacat cggagagctt tttttttacg cgt             593


<210>  372
<211>  593
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD1.dcasffok.pamin.3

<400>  372
acgcgtaaaa aaaaagggct ggctgcggtc ctcgggtccc ctcgtgaggg gcaatcgttg       60

ggtgtttcgt ggtctcatac agaacttata agattcccaa atccaaagac atttcacgtt      120

tatggtgatt tcccagaaca catagcgaca tgcaaatatt gcagggcgcc actcccctgt      180

ccctcacagc catcttcctg ccagggcgca cgcgcgctgg gtgttcccgc ctagtgacac      240

tgggcccgcg attccttgga gcgggttgat gacgtcagcg ttcaaagagg gcctatttcc      300

catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa      360

tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct      420

tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac      480

ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacccaacg      540

attgcccctc acgaggggac ttgctggggc tcatgcggta tttttttacg cgt             593


<210>  373
<211>  11080
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<223>  human MPO gene

       Accession Number: NC_000017.11>NC_000017.11:c58280935-58269856 
       Homo sapiens chromosome 17, GRCh38.p12 Primary Assembly

<400>  373
gacaatatca ggtgagctgt ggaggtgggg tccttggaag ctggatgaca gcagctggca       60

aggggataag agagcagtga gcccctccct caaggaggtc tggctttatc catagacagg      120

gccctctgag gtggggctga ggtacaaagg gggattgagc agcccaggag aagagagatg      180

ggggttccct tcttctcttc tctcagatgc atggtggact taggaccttg ctgggctggg      240

ggtctcactg cagagatgaa gctgcttctg gccctagcag ggctcctggc cattctggcc      300

acgccccagc cctctgaagg tgctgctcca ggtaacagtt cccaaggtgg gagaagatgg      360

tgtgttgggg tggttgtgtt ccagagaccc cttttctcag agcaggcctt cctagctctg      420

gggcctgata gggggttggg gcccattcct gactttgtga tccctgctct gggcagctgt      480

cctgggggag gtggacacct cgttggtgct gagctccatg gaggaggcca agcagctggt      540

ggacaaggcc tacaaggagc ggcgggaaag gtggggcacg aggcactgcc cagctctggg      600

caaggatgtc ccaggccttt cagagaagca gcaggcagca gggagcttga aggtgggaga      660

gggggactat ggggacccat gtggactccc tttctgtctg tctgtccctg tgtttctgga      720

tgggagaaca gcttaaaagc catccttccc aaagccttgc ctctgtctgg acacccaggt      780

ccatatcctt tttataaaag gaagccttcc tcccacccgt gggcctgcct gcatggtcct      840

gcatgtctgc ccctgggctg gggctctgct gggtgggtct gtatcccttc tctggccctc      900

actcctcctt tctccaagca gcatcaagca gcggcttcgc agcggctcag ccagccccat      960

ggaactccta tcctacttca agcagccggt ggcagccacc aggacggcgg tgagggccgc     1020

tgactacctg cacgtggctc tagacctgct ggagaggaag ctgcggtccc tgtggcgaag     1080

gccattcaat gtcactggta ctgttgccca tcacacccca ggtccctgct ccactagtga     1140

ctcctcctag ggaccccagc cgtctctcag tgatcccctc aacttcttcc tccagggagt     1200

ctcaggagtc tccagggctc ctcagcctca ggttgcctgg gataggaagt gaggcggctc     1260

agctccccca ttgttctttc ccccggcaga tgtgctgacg cccgcccagc tgaatgtgtt     1320

gtccaagtca agcggctgcg cctaccagga cgtgggggtg acttgcccgg agcaggacaa     1380

ataccgcacc atcaccggga tgtgcaacaa caggtgcggc tggctggggg tggctgcagg     1440

aaccgggctc agagaggcgt cccggacgcc acaagcctcc cggtgtcagc gccctgtcct     1500

cccctgcaga cgcagcccca cgctgggggc ctccaaccgt gcctttgtgc gctggctgcc     1560

ggcggagtat gaggacggct tctctcttcc ctacggctgg acgcccgggg tcaagcgcaa     1620

cggcttcccg gtggctctgg tgagcgccgg cgggcagagg gggcgaggcc cggccacgcg     1680

gtgcgcggac ccaggcgcca gctgatctcc gtgtcccgca ggctcgcgcg gtctccaacg     1740

agatcgtgcg cttccccact gatcagctga ctccggacca ggagcgctca ctcatgttca     1800

tgcaatgggg ccagctgttg gaccacgacc tcgacttcac ccctgagccg gccgcccggg     1860

cctccttcgt cactggcgtc aactgcgaga ccagctgcgt tcagcagccg ccctgcttcc     1920

cgctcaaggt ggccctgctt cccgcccact gcctgggttg ggagagggga gattgtttct     1980

ggaaggggcc atctcccttc tgtgcccagg tttcctttcc cagacgctga ggaaggctgg     2040

ccctgcctcc cttgtgtcca cagcactggc tgctcagcta tgacctgtct ccttcctgcg     2100

cctgggctca gccagctggc agccagggcc gtccatttgc tcactgcctc tggcaggcca     2160

agggagagag ggttgttcct ccttggctga caggagtcct gttggtggaa tcactggctc     2220

tttgttaggt tgggagaggg ttctgggagg agaccttctc agtgcccccc aagcctccat     2280

gctcagtcct gtttccgccc cactgagagg gcagccccaa cccccatcac acagaaagag     2340

agagcctggg ggtctggtgg gtgccgtggg acagggctgg gagtgggcag ggtggtcgag     2400

gcactgctga gtagggaagg aggaggagag aaaggagaga gcgcctgaga gagggagaga     2460

cagacaggag aatgtcaggg ccagagggag agcaggcaca gcagagagaa ggggagagag     2520

acgggcgacg ctttagtgag gagggatccg aggcctggcg gtgcccagac ccaggggttg     2580

cccaggttcc cagttcagtg ttctgctcat taaccctgca cctcagaggc tgttgctgat     2640

ggcgccctag gcagcgatcc ctttcgggcc tccggaggcc tctctgcagg ttgagggtac     2700

cagaggtcct gagggcaggg ggagatccag ttctgcctgg gcaccttcct tgccctcggt     2760

gagccagtct agcctctctc tgtgcctcag atcccgccca atgacccccg catcaagaac     2820

caagccgact gcatcccgtt cttccgctcc tgcccggctt gccccgggag caacatcacc     2880

atccgcaacc agatcaacgc gctcacttcc ttcgtggacg ccagcatggt gtacggcagc     2940

gaggagcccc tggccaggaa cctgcgcaac atgtccaacc agctggggct gctggccgtc     3000

aaccagcgct tccaagacaa cggccgggcc ctgctgccct ttgacaacct gcacgatgac     3060

ccctgtctcc tcaccaaccg ctcagcgcgc atcccctgct tcctggcagg tcagctttgg     3120

ggtggggacc agaggtggca taggagatgt tccctgttgg agccacagtg agtctgtttg     3180

tgagcagctt gtgggtttgt acttagagag actgtcctca ccagccatta ctatcaactt     3240

catgatatta atccagttgc cttggtaaca agtcagatgg agccagaaaa gcagaaaagc     3300

aaacaacctc tccaacccta agatggagac ccaggcatgg ctgaggagcc tctgtccatc     3360

agtccattgc tttctctgtc ccttttttcc cttctggact ctggaagaca agagagtcaa     3420

atccctcagg ggcagggaac taaaggacag agaacgctgt ggtgccctca ccctctagag     3480

cgacactatc caatgtggta gccactagct gcatgtggct attttaattt tagttatttt     3540

aaattaaata aacccagttt ctcagcggtg ctaaccacat ttcaaatgct caattgcctt     3600

acagtctagt ggctccccct attgggtggc acagatgtaa aatattttca tcatctcgga     3660

aggttgagtt ggaccacact gctccaaaat atcccctcct gcgcatctta attacccata     3720

ataagagatg atgggatttt aaatgtaaac aattggcagt tttggagaca caattggggc     3780

agtgatggca gccatgacca gacattttcc agtgtttttt ttttttctaa gacggaatct     3840

cactctgtca cccaggcttg agtacagtgg cacagtctcg gctcactgca acctctgcca     3900

actgggttta agtgattctt ctgcctcagc ctcctgagta gctgggacta caggcgcctg     3960

ccaccacact tggctaattt tttgtatttt tagtagagat ggtgttttac catgttggcc     4020

aggctggtct caaactcctg acctcatgat ctgcccacct taacctccca aagtgctggg     4080

attacaggtg tgagccaccg cacgcagcca ttttccagtg tttttaaatg taaacccaat     4140

agtggacctc aggccctttc ctctggcctc cctctgcctc ctcctgatgg ttcctgctgg     4200

tctctccctg gctgggaccc tgggaggagg atccaggtcg gactttctgc aagagatgcc     4260

cgacctcttc tgggctagtt aggcaagggc agccctcctt agccagactc agctgcccac     4320

agcctggctt tgatgagccc catgtcccaa gcctgggtca ttatgacctc tagctgttca     4380

tggatggaga tgctgctctc agagaatttc ctttccacca gcttctagag ctggcaagga     4440

ctgaccctag tgtcctgacc ctagccctct ggcccctgca gagaagggtt ctgggaggac     4500

taggggaacc aggggggatc tgcttagttt tcttgaacag tctcaggtgt ggactccaca     4560

tatctcatgt cctagccctc tcccctgggg caggctgcac cattggcatg tagtaaaggg     4620

cagggctgac aactctgttc aaatggtagt ggctttctgg agtcctgtgt tgagaaggat     4680

tctgaggttg catccaattt tagcaggaaa gagtgctgtg atcgttagct gcatcagaca     4740

tgggcatgga tgtaggctgt gccaacacgc atgcttcttc ccagacactc cttggaatca     4800

agagaatcaa tgaaccaagc agtgttcata agtctcatac tttgtaccag gcactctgtt     4860

aagccccatg tatccatatg tgtgctagga gggagggtaa aatataaaaa ggcacaagcc     4920

ttggtcctgt gctcaaggag ctcccagttt caggtacagg aaagtggcat gtcatgtgtt     4980

ggaagtgata cctattgaat gagcatcccc aaccagcagc ggccctgagc tgaggcagaa     5040

gctatcattg tggactgggg catcagggaa ggcctcctag aggagtgtaa attggggata     5100

gtttgaaaag atggatgggg aggaggggtg agtaggaggc atttcaggcc ttggctgggg     5160

agggggtttc agtggagcaa atcttttctg ggatggaggc cttaagaatg acagtggctt     5220

ttgcctcccc aaggggacac ccgttccagt gagatgcccg agctcacctc catgcacacc     5280

ctcttacttc gggagcacaa ccggctggcc acagagctca agagcctgaa ccctaggtgg     5340

gatggggaga ggctctacca ggaagcccgg aagatcgtgg gggccatggt ccaggtaggc     5400

cgtcttgaac accgggcaca cgggatccag atgtgtccct gcaacatcct agctgtgtga     5460

ccttgggcaa gttcctaacg ctcctgagcc ttgccttctt catccatgag gctgtaagga     5520

ttataataca tatatgaaaa cacctttcag atagtaagtg cttgacaaag cctccttccc     5580

ttccctctcc ttcttcgcag gtttgctccc tgcagtcttc tttcctgtgg gtctgttatt     5640

ggtgttggga gggttgaatg tttggtttag cgcccaagta gctgttcagt aaattcttct     5700

tccctccatc tcactgtcac tcttagctcc tttacccact cacctcttct ttttaaaatt     5760

aaaaaaatta tagggctggg cgcggtggct catgcctgta atcccagcac tttgggaggc     5820

ggaggcgggt ggatcacgag gtcaggagat caagaccatg ctggctaaca cggtgaaacc     5880

ccatctctac taaaaataca aaaattagcc gggcgtggtg gcgggcacct gtagtcccag     5940

ctacttggga ggctgaggca ggagaatggc gtgaacctgg gaggcagagc ttgcagtgag     6000

ctgagatggc gctgctgcac tccagcctgg gtgacagagc gagactccat ctcaaaataa     6060

ataaataaat aaaataaaat ttaaaaatta ttattattat tttattttta ttttttgata     6120

cagggtctta ccctgttgcc caggctggag tgcagtggtg cgatcttggc tcactgcaac     6180

ctccaccttt caggctcaag ccattctccc atctcagcct tctgagtagc tgggatttca     6240

agcacgggcc accacgcccg gctaattttt gtattttttg tagagacggg gtttcgccac     6300

gttacccagt gtggtctcaa actcctgggc tcaagtaatc cgccctcctc gacctcccaa     6360

atccctggga ttacaggcac gagtcactgt gttcagccca ctcacctctt ctttttggct     6420

ctgtcataat ccacccaatg tccttagcct catttacccc tggggcaatg ttcctggttc     6480

tgtctctcaa catctggtgc ttcattgccc ccaccctctc cctcagaggc agaggcagac     6540

atacacacac tcacacacac acacacacac acacacacac acacacactc ctagattctg     6600

gaggagagag ttttcggtgc taagggtgag tttttgattt gcactaagaa ctcaagtgaa     6660

attcatttac actccaaaat aacttctaga ctctagaatg tagagtgaga ctattccatg     6720

tcttcacttt tcacagggaa aatcatgcca gaatggccct tcgttccttc ctgcctccct     6780

gctttccaca aacctaaact gagccttact gtgggcaggc acttggtggg cactgtgaag     6840

aatttcaaca acttcaattt cccttgctct caaggacttt ataagatagt tgcagaaatg     6900

agacattaat aacttgagga caaggaagtt atcacaatga ctaaaaaagt gatttgctat     6960

ttagtcctct aaggcagagg gcaatataat tatccttaat gtacatttga ggaactcagg     7020

ctcagagagg ttaataattg gtccaaggtc acccagccag caagcagtag aactctattt     7080

atgcctctca gtgccactct gttaggccca ggtccggtat tgtgggaggc tattccctga     7140

ccatagaggg atggaagggg tccccgaagc caagagcagg cagagactct ggccctcctg     7200

tgctgccagg cggcatttgc tgtggaggag tgggtggaga ggccattcca aatgacttgt     7260

ctttagatca tcacttaccg ggactacctg cccctggtgc tggggccaac ggccatgagg     7320

aagtacctgc ccacgtaccg ttcctacaat gactcagtgg acccacgcat cgccaacgtc     7380

ttcaccaatg ccttccgcta cggccacacc ctcatccaac ccttcatgtt ccgcctggac     7440

aatcggtacc agcccatgga acccaacccc cgtgtccccc tcagcagggt cttttttgcc     7500

tcctggaggg tcgtgctgga aggtaagcag gacctaggcc aggagcgagt gggctgacag     7560

ctaaaggtgg ggtagggatc accttggctc tagggagcta gggtggggag cagtctgctt     7620

tcctcccatc ttggagatct ggtctgactc tctagcctca gttcctctga ctctgcttcc     7680

atatctgata ataaacagtg gggtggggat ggccctcaga gggaactggc tacttcctat     7740

gtgttgtggc tcgctgtgac ccaggaagcc agggcctggg agcatcccaa tcttgcaagt     7800

ctgaatggca taatctcctg agacctatat gtccctgggg tcatgtagtg accggtgagg     7860

acctgctcac tggggccacc accttggcac caggcactgc ttgtgcccag ccctcttctc     7920

gaatcctcct gaccctacct ggacttgtcc ctgactccaa tctgagctct gatactgagc     7980

cagatacttc ccctgacctg gctcctctgg tccccaggtg gcattgaccc catcctccgg     8040

ggcctcatgg ccacccctgc caagctgaat cgtcagaacc aaattgcagt ggatgagatc     8100

cgggagcgat tgtttgagca ggtcatgagg attgggctgg acctgcctgc tctgaacatg     8160

cagcgcagca gggaccacgg cctcccaggt gaggggctgc aggagtctcc cctgatgctc     8220

acctcccctg agctgcctcc taggtagccc atcacccccc ttcccactct agggtccctg     8280

ccctctccaa agcatattag aggcactttg ctctcaggaa gtagcctgag gccaggcccc     8340

tgccttttcc cggacccacc tgataaattt ccagctcctc tcaacctcct ttgcatgctg     8400

taattctcat attaggggca tcgtcatcat aatgataacc cacagtctga aaatctgtct     8460

ctgtcatcat tccattttgc ttcattgtac gttaactctg ggggaagaga ttattattgc     8520

cttcatttta ctgatgagga aactgaggct tgatgagatt cagaacctgg ccaggttctc     8580

tagccaagtc agtggcaaag ctgaaatttg aagccagatt agcctaacct caaagcctgt     8640

gcctatgtta tggggtagcc catgtggctg aggcttctag acacatttca cctctcatgg     8700

gtcagagaca gacacatcgg gctgctctgg agggggacaa ggagatgtcc ctattcttgg     8760

acccagcagg aactgtggca gtttagggtt tgccttatgg taggagggca agagaaagac     8820

agagagggtc ccagagaggc ccagttgtgc cccttggtct aagccaaagg tacagggttc     8880

cttgaggtga ggtccttccc tccctctatg accttggttg tgttgggaag gtgggttttc     8940

tgggctgctg gcgatatgtg aatcttccct gctgtctcca gtgacctccc caccttgaag     9000

cagagagacc tgcccatagc cacctgtccc ctcccccatg caggatacaa tgcctggagg     9060

cgcttctgtg ggctcccgca gcctgaaact gtgggccagc tgggcacggt gctgaggaac     9120

ctgaaattgg cgaggaaact gatggagcag tatggcacgc ccaacaacat cgacatctgg     9180

atgggcggcg tgtccgagcc tctgaagcgc aaaggccgcg tgggcccact cctcgcctgc     9240

atcatcggta cccagttcag gaagctccgg gatggtgatc ggtgaggagg ggcaggcgtc     9300

gtgggccgct gggtggctgt gggcccatcc ttgactctct tggagcccaa atttcctcct     9360

gtcagttgaa ggactggagg gagtcagtaa ttttccagtg tgttccagga atcctccaga     9420

tgccctggag tgcccctaat gtgccctgaa cctgttgggg gtgcagcaag gaaagacctg     9480

gacccctggt tgtaaagggt ggaagataga aaagtgccac ccttagctac ttcttcccat     9540

gggtttcact gtgccaccat gtgggtgtca gtcaccagtg ggcagtcccc tcccagcttt     9600

cccacagctc cccttagtgt catttcctcg tggtcctgat gtcttcatgt ctagttcttg     9660

cttagcctgt actgcacctt tgggaaaatt agaaaaaggt gcccctctct gggtgggcga     9720

attataaacc acccccaaga ttctcttcca ctcctgtgga tgcagctcct ggccagggca     9780

cagggcttca ggagtgtgtg tgggcccggg tccccctcca gcatggtgcc ctggaggccg     9840

ccgggctagg ggtaagggaa aggccactgg gcagctgtgc tttactctgc acgatgagtg     9900

gagcatcact tgtgtgaaag cccctgggct gcccaagggc ctggggccct cctgtgcttc     9960

caggtggcat ttgttgtggc tttgttatat cctgggagca gcacaagccc atcgatgccc    10020

tgccagccca gaatatcctt gggcacagtg tccatgggtg ttccccatgc aggttttggt    10080

gggagaacga gggtgtgttc agcatgcagc agcgacaggc cctggcccag atctcattgc    10140

cccggatcat ctgcgacaac acaggcatca ccaccgtgtc taagaacaac atcttcatgt    10200

ccaactcata tccccgggac tttgtcaact gcagtacact tcctgcattg aacctggctt    10260

cctggaggga agcctcctag aggccaggta agggggtgca gcagtgaggg gtatatctgg    10320

gctggccagt tggaaccacg gagatctcct tgccctagat gagcccagcc ctgttctggg    10380

tgcagctgag aaaatgagtg actagacgtt catttgtgtg ctcatgtatg tgcgaagtat    10440

ataaattggc ttttcatgcg tgtgtgttgt ctgaacatgg ggagtgtttc atgggttatg    10500

tgtatgtgcc atttatgtga gtgtgtgttt gtgctgatga gaatactgag tatgtggaag    10560

gcagcagagc ggactggtga ggagcacagc tcaggaacta gactgcctgg gttccaatcc    10620

tggctctgtg gcttgctagc tatgtgacct tgagcaaatt accctcctta aacaagagtt    10680

ttcttccttg taaattacat ctgtcatggt ttcttggagg gcccacttgt atcctctggt    10740

tcttcattta ttgagcacct actacatgca aggcactgta ctaggcgtga gaagcatata    10800

gaggcaagaa agagatacca agatgccatc tgtgtcctgg ttagcagagc tggaccagtg    10860

gtgccttgga gggataagcc agctgcagct gggctgtgtg gttgacttat gggcccagcc    10920

agccaggctc aggccatggc tccccttttt cttcctcacc ctgatttctt gcttattcac    10980

tgaagttctc ctgaagagga actgggcctg ttgccctttc tgtaccattt atttgctccc    11040

aatgtttatg ataataaagg caccgctgat ggggacctcc                          11080


<210>  374
<211>  745
<212>  PRT
<213>  Homo sapiens


<220>
<221>  MISC_FEATURE
<223>  MPO protein Accession Number: NP_000241

<400>  374

Met Gly Val Pro Phe Phe Ser Ser Leu Arg Cys Met Val Asp Leu Gly 
1               5                   10                  15      


Pro Cys Trp Ala Gly Gly Leu Thr Ala Glu Met Lys Leu Leu Leu Ala 
            20                  25                  30          


Leu Ala Gly Leu Leu Ala Ile Leu Ala Thr Pro Gln Pro Ser Glu Gly 
        35                  40                  45              


Ala Ala Pro Ala Val Leu Gly Glu Val Asp Thr Ser Leu Val Leu Ser 
    50                  55                  60                  


Ser Met Glu Glu Ala Lys Gln Leu Val Asp Lys Ala Tyr Lys Glu Arg 
65                  70                  75                  80  


Arg Glu Ser Ile Lys Gln Arg Leu Arg Ser Gly Ser Ala Ser Pro Met 
                85                  90                  95      


Glu Leu Leu Ser Tyr Phe Lys Gln Pro Val Ala Ala Thr Arg Thr Ala 
            100                 105                 110         


Val Arg Ala Ala Asp Tyr Leu His Val Ala Leu Asp Leu Leu Glu Arg 
        115                 120                 125             


Lys Leu Arg Ser Leu Trp Arg Arg Pro Phe Asn Val Thr Asp Val Leu 
    130                 135                 140                 


Thr Pro Ala Gln Leu Asn Val Leu Ser Lys Ser Ser Gly Cys Ala Tyr 
145                 150                 155                 160 


Gln Asp Val Gly Val Thr Cys Pro Glu Gln Asp Lys Tyr Arg Thr Ile 
                165                 170                 175     


Thr Gly Met Cys Asn Asn Arg Arg Ser Pro Thr Leu Gly Ala Ser Asn 
            180                 185                 190         


Arg Ala Phe Val Arg Trp Leu Pro Ala Glu Tyr Glu Asp Gly Phe Ser 
        195                 200                 205             


Leu Pro Tyr Gly Trp Thr Pro Gly Val Lys Arg Asn Gly Phe Pro Val 
    210                 215                 220                 


Ala Leu Ala Arg Ala Val Ser Asn Glu Ile Val Arg Phe Pro Thr Asp 
225                 230                 235                 240 


Gln Leu Thr Pro Asp Gln Glu Arg Ser Leu Met Phe Met Gln Trp Gly 
                245                 250                 255     


Gln Leu Leu Asp His Asp Leu Asp Phe Thr Pro Glu Pro Ala Ala Arg 
            260                 265                 270         


Ala Ser Phe Val Thr Gly Val Asn Cys Glu Thr Ser Cys Val Gln Gln 
        275                 280                 285             


Pro Pro Cys Phe Pro Leu Lys Ile Pro Pro Asn Asp Pro Arg Ile Lys 
    290                 295                 300                 


Asn Gln Ala Asp Cys Ile Pro Phe Phe Arg Ser Cys Pro Ala Cys Pro 
305                 310                 315                 320 


Gly Ser Asn Ile Thr Ile Arg Asn Gln Ile Asn Ala Leu Thr Ser Phe 
                325                 330                 335     


Val Asp Ala Ser Met Val Tyr Gly Ser Glu Glu Pro Leu Ala Arg Asn 
            340                 345                 350         


Leu Arg Asn Met Ser Asn Gln Leu Gly Leu Leu Ala Val Asn Gln Arg 
        355                 360                 365             


Phe Gln Asp Asn Gly Arg Ala Leu Leu Pro Phe Asp Asn Leu His Asp 
    370                 375                 380                 


Asp Pro Cys Leu Leu Thr Asn Arg Ser Ala Arg Ile Pro Cys Phe Leu 
385                 390                 395                 400 


Ala Gly Asp Thr Arg Ser Ser Glu Met Pro Glu Leu Thr Ser Met His 
                405                 410                 415     


Thr Leu Leu Leu Arg Glu His Asn Arg Leu Ala Thr Glu Leu Lys Ser 
            420                 425                 430         


Leu Asn Pro Arg Trp Asp Gly Glu Arg Leu Tyr Gln Glu Ala Arg Lys 
        435                 440                 445             


Ile Val Gly Ala Met Val Gln Ile Ile Thr Tyr Arg Asp Tyr Leu Pro 
    450                 455                 460                 


Leu Val Leu Gly Pro Thr Ala Met Arg Lys Tyr Leu Pro Thr Tyr Arg 
465                 470                 475                 480 


Ser Tyr Asn Asp Ser Val Asp Pro Arg Ile Ala Asn Val Phe Thr Asn 
                485                 490                 495     


Ala Phe Arg Tyr Gly His Thr Leu Ile Gln Pro Phe Met Phe Arg Leu 
            500                 505                 510         


Asp Asn Arg Tyr Gln Pro Met Glu Pro Asn Pro Arg Val Pro Leu Ser 
        515                 520                 525             


Arg Val Phe Phe Ala Ser Trp Arg Val Val Leu Glu Gly Gly Ile Asp 
    530                 535                 540                 


Pro Ile Leu Arg Gly Leu Met Ala Thr Pro Ala Lys Leu Asn Arg Gln 
545                 550                 555                 560 


Asn Gln Ile Ala Val Asp Glu Ile Arg Glu Arg Leu Phe Glu Gln Val 
                565                 570                 575     


Met Arg Ile Gly Leu Asp Leu Pro Ala Leu Asn Met Gln Arg Ser Arg 
            580                 585                 590         


Asp His Gly Leu Pro Gly Tyr Asn Ala Trp Arg Arg Phe Cys Gly Leu 
        595                 600                 605             


Pro Gln Pro Glu Thr Val Gly Gln Leu Gly Thr Val Leu Arg Asn Leu 
    610                 615                 620                 


Lys Leu Ala Arg Lys Leu Met Glu Gln Tyr Gly Thr Pro Asn Asn Ile 
625                 630                 635                 640 


Asp Ile Trp Met Gly Gly Val Ser Glu Pro Leu Lys Arg Lys Gly Arg 
                645                 650                 655     


Val Gly Pro Leu Leu Ala Cys Ile Ile Gly Thr Gln Phe Arg Lys Leu 
            660                 665                 670         


Arg Asp Gly Asp Arg Phe Trp Trp Glu Asn Glu Gly Val Phe Ser Met 
        675                 680                 685             


Gln Gln Arg Gln Ala Leu Ala Gln Ile Ser Leu Pro Arg Ile Ile Cys 
    690                 695                 700                 


Asp Asn Thr Gly Ile Thr Thr Val Ser Lys Asn Asn Ile Phe Met Ser 
705                 710                 715                 720 


Asn Ser Tyr Pro Arg Asp Phe Val Asn Cys Ser Thr Leu Pro Ala Leu 
                725                 730                 735     


Asn Leu Ala Ser Trp Arg Glu Ala Ser 
            740                 745 


<210>  375
<211>  1625
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartite SV40 NLS (TG14280)

<400>  375

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  
    1625 


<210>  376
<211>  1631
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartite SV40 NLS, 6xHis (TG14412)

<400>  376

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  His His His His His  His 
    1625                 1630     


<210>  377
<211>  1625
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2  domain, SV40 and bipartite SV40
       NLS (TG14314)

<400>  377

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  
    1625 


<210>  378
<211>  1506
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta, SV40+bipartiteSV40 NLS (TG14297)

<400>  378

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1430                 1435                 1440             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1445                 1450                 1455             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1460                 1465                 1470             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Ser  Gly Gly Ser 
    1475                 1480                 1485             


Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu Ser Pro  Lys Lys Lys 
    1490                 1495                 1500             


Arg Lys  Val 
    1505     


<210>  379
<211>  1513
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with longer linker, SV40+bipartiteSV40 NLS 
       (TG14298)

<400>  379

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1505                 1510             


<210>  380
<211>  1616
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNH replaced with SSB, SV40+bipartiteSV40 NLS 
       (TG14299)

<400>  380

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Thr Thr  Glu Thr Phe Val Lys  Asp Ile Lys Pro Gly  Leu Lys Asn 
    1025                 1030                 1035             


Leu Asn  Leu Ile Phe Ile Val  Leu Glu Thr Gly Arg  Val Thr Lys 
    1040                 1045                 1050             


Thr Lys  Asp Gly His Glu Val  Arg Thr Cys Lys Val  Ala Asp Lys 
    1055                 1060                 1065             


Thr Gly  Ser Ile Asn Ile Ser  Val Trp Asp Asp Val  Gly Asn Leu 
    1070                 1075                 1080             


Ile Gln  Pro Gly Asp Ile Ile  Arg Leu Thr Lys Gly  Tyr Ala Ser 
    1085                 1090                 1095             


Val Phe  Lys Gly Cys Leu Thr  Leu Tyr Thr Gly Arg  Gly Gly Asp 
    1100                 1105                 1110             


Leu Gln  Lys Ile Gly Glu Phe  Cys Met Val Tyr Ser  Glu Val Pro 
    1115                 1120                 1125             


Asn Phe  Ser Glu Pro Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Ala Thr  Val Arg Lys Val Leu  Ala Met Pro Gln Val  Asn Ile Val 
    1310                 1315                 1320             


Lys Lys  Thr Glu Val Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile 
    1325                 1330                 1335             


Leu Ser  Lys Arg Glu Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly 
    1340                 1345                 1350             


Trp Asp  Thr Arg Lys Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala 
    1355                 1360                 1365             


Tyr Ser  Ile Leu Val Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys 
    1370                 1375                 1380             


Lys Leu  Lys Ser Val Lys Val  Leu Val Gly Ile Thr  Ile Met Glu 
    1385                 1390                 1395             


Lys Gly  Ser Tyr Glu Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys 
    1400                 1405                 1410             


Gly Tyr  Lys Asp Ile Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys 
    1415                 1420                 1425             


Tyr Ser  Leu Phe Glu Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala 
    1430                 1435                 1440             


Ser Ala  Thr Glu Leu Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln 
    1445                 1450                 1455             


His Leu  Val Arg Leu Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr 
    1460                 1465                 1470             


Thr Gly  Ser Asn Asn Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu 
    1475                 1480                 1485             


Phe Lys  Glu Ile Phe Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr 
    1490                 1495                 1500             


Ile Leu  Lys Asn Lys Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp 
    1505                 1510                 1515             


Glu Gln  Phe Ala Val Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe 
    1520                 1525                 1530             


Val Ser  Leu Leu Lys Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe 
    1535                 1540                 1545             


Thr Phe  Leu Asp Leu Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln 
    1550                 1555                 1560             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1565                 1570                 1575             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1580                 1585                 1590             


Asp Ser  Gly Gly Ser Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu 
    1595                 1600                 1605             


Ser Pro  Lys Lys Lys Arg Lys  Val 
    1610                 1615     


<210>  381
<211>  1488
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with longer deletion SV40+bipartiteSV40 NLS 
       (TG14300)

<400>  381

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Gly Gly Ser  Gly Gly Leu 
        995                 1000                 1005             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1010                 1015                 1020             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1025                 1030                 1035             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1040                 1045                 1050             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1055                 1060                 1065             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1070                 1075                 1080             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1085                 1090                 1095             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1100                 1105                 1110             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1115                 1120                 1125             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1130                 1135                 1140             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1145                 1150                 1155             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1160                 1165                 1170             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1175                 1180                 1185             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1190                 1195                 1200             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1205                 1210                 1215             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1220                 1225                 1230             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1235                 1240                 1245             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1250                 1255                 1260             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1265                 1270                 1275             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1280                 1285                 1290             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1295                 1300                 1305             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1310                 1315                 1320             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1325                 1330                 1335             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1340                 1345                 1350             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1355                 1360                 1365             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1370                 1375                 1380             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1385                 1390                 1395             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1400                 1405                 1410             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1415                 1420                 1425             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1430                 1435                 1440             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1445                 1450                 1455             


Asp Leu  Ser Gln Leu Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr 
    1460                 1465                 1470             


Ala Asp  Gly Ser Glu Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1475                 1480                 1485             


<210>  382
<211>  1513
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with longer positively charged linker, 
       SV40+bipartiteSV40 NLS (TG14301)

<400>  382

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Lys 
    1010                 1015                 1020             


Ala Arg  Gly Lys Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1505                 1510             


<210>  383
<211>  1506
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with shorter positively charged linker, 
       SV40+bipartiteSV40 NLS (14302)

<400>  383

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Lys Ser 
    1010                 1015                 1020             


Lys Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1430                 1435                 1440             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1445                 1450                 1455             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1460                 1465                 1470             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Ser  Gly Gly Ser 
    1475                 1480                 1485             


Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu Ser Pro  Lys Lys Lys 
    1490                 1495                 1500             


Arg Lys  Val 
    1505     


<210>  384
<211>  1580
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNH replaced with StkC, SV40+bipartiteSV40 NLS 
       (TG14303)

<400>  384

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ser Val  Ser Met Val His Ser  Glu Arg Leu Pro Leu  Leu Met Asn 
    1025                 1030                 1035             


Arg Asn  Gly Ile Arg Phe Gln  Ser Glu Val Ile Asp  Pro Asn Pro 
    1040                 1045                 1050             


Pro Lys  Lys Val Lys Trp Val  Leu Thr Gly Ser Gly  Gly Asn Met 
    1055                 1060                 1065             


Phe Thr  Ala Asn Val Arg Gly  Ser Asn Val Ser Ser  Leu Val Lys 
    1070                 1075                 1080             


His Thr  Trp Ser Glu Ala Phe  Val Ser Leu Ile Glu  Asp Arg Gly 
    1085                 1090                 1095             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1100                 1105                 1110             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1115                 1120                 1125             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1130                 1135                 1140             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1145                 1150                 1155             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1160                 1165                 1170             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1175                 1180                 1185             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1190                 1195                 1200             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1205                 1210                 1215             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1220                 1225                 1230             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1235                 1240                 1245             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1250                 1255                 1260             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1265                 1270                 1275             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1280                 1285                 1290             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1295                 1300                 1305             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1310                 1315                 1320             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1325                 1330                 1335             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1340                 1345                 1350             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1355                 1360                 1365             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1370                 1375                 1380             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1385                 1390                 1395             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1400                 1405                 1410             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1415                 1420                 1425             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1430                 1435                 1440             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1445                 1450                 1455             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1460                 1465                 1470             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1475                 1480                 1485             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1490                 1495                 1500             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1505                 1510                 1515             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1520                 1525                 1530             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1535                 1540                 1545             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1550                 1555                 1560             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1565                 1570                 1575             


Lys Val  
    1580 


<210>  385
<211>  1499
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, HNHdelta with HNS linker, SV40+bipartiteSV40 NLS 
       (TG14304)

<400>  385

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Ala Ala Val  Lys Ser Gly 
        995                 1000                 1005             


Thr Lys  Ala Lys Arg Ala Gln  Arg Pro Gly Leu Ser  Glu Ala Asp 
    1010                 1015                 1020             


Lys Ala  Gly Phe Ile Lys Arg  Gln Leu Val Glu Thr  Arg Gln Ile 
    1025                 1030                 1035             


Thr Lys  His Val Ala Arg Ile  Leu Asp Ser Arg Met  Asn Thr Lys 
    1040                 1045                 1050             


Arg Asp  Lys Asn Asp Lys Pro  Ile Arg Glu Val Lys  Val Ile Thr 
    1055                 1060                 1065             


Leu Lys  Ser Lys Leu Val Ser  Asp Phe Arg Lys Asp  Phe Gln Leu 
    1070                 1075                 1080             


Tyr Lys  Val Arg Asp Ile Asn  Asn Tyr His His Ala  His Asp Ala 
    1085                 1090                 1095             


Tyr Leu  Asn Ala Val Val Gly  Thr Ala Leu Ile Lys  Lys Tyr Pro 
    1100                 1105                 1110             


Lys Leu  Glu Ser Glu Phe Val  Tyr Gly Asp Tyr Lys  Val Tyr Asp 
    1115                 1120                 1125             


Val Arg  Lys Met Ile Ala Lys  Ser Glu Gln Glu Ile  Gly Lys Ala 
    1130                 1135                 1140             


Thr Ala  Lys Arg Phe Phe Tyr  Ser Asn Ile Met Asn  Phe Phe Lys 
    1145                 1150                 1155             


Thr Glu  Val Lys Leu Ala Asn  Gly Glu Ile Arg Lys  Arg Pro Leu 
    1160                 1165                 1170             


Ile Glu  Thr Asn Gly Glu Thr  Gly Glu Val Val Trp  Asn Lys Glu 
    1175                 1180                 1185             


Lys Asp  Phe Ala Thr Val Arg  Lys Val Leu Ala Met  Pro Gln Val 
    1190                 1195                 1200             


Asn Ile  Val Lys Lys Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys 
    1205                 1210                 1215             


Glu Ser  Ile Leu Ser Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg 
    1220                 1225                 1230             


Lys Lys  Gly Trp Asp Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro 
    1235                 1240                 1245             


Thr Val  Ala Tyr Ser Ile Leu  Val Val Ala Lys Val  Glu Lys Gly 
    1250                 1255                 1260             


Lys Ala  Lys Lys Leu Lys Ser  Val Lys Val Leu Val  Gly Ile Thr 
    1265                 1270                 1275             


Ile Met  Glu Lys Gly Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu 
    1280                 1285                 1290             


Glu Ala  Lys Gly Tyr Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys 
    1295                 1300                 1305             


Leu Pro  Lys Tyr Ser Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg 
    1310                 1315                 1320             


Met Leu  Ala Ser Ala Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val 
    1325                 1330                 1335             


Leu Pro  Gln His Leu Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile 
    1340                 1345                 1350             


Ser Ala  Thr Thr Gly Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His 
    1355                 1360                 1365             


Arg Glu  Glu Phe Lys Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser 
    1370                 1375                 1380             


Glu Lys  Tyr Ile Leu Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser 
    1385                 1390                 1395             


Ser Phe  Asp Glu Gln Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser 
    1400                 1405                 1410             


Asn Ser  Phe Val Ser Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Ser 
    1415                 1420                 1425             


Gly Gly  Phe Thr Phe Leu Asp  Leu Asp Val Lys Gln  Gly Arg Leu 
    1430                 1435                 1440             


Arg Tyr  Gln Thr Val Thr Glu  Val Leu Asp Ala Thr  Leu Ile Tyr 
    1445                 1450                 1455             


Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg Thr Asp  Leu Ser Gln 
    1460                 1465                 1470             


Leu Gly  Gly Asp Ser Gly Gly  Ser Lys Arg Thr Ala  Asp Gly Ser 
    1475                 1480                 1485             


Glu Phe  Glu Ser Pro Lys Lys  Lys Arg Lys Val 
    1490                 1495                 


<210>  386
<211>  1506
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNHdelta, SV40+bipartiteSV40 
       NLS (14315)

<400>  386

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1430                 1435                 1440             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1445                 1450                 1455             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1460                 1465                 1470             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Ser  Gly Gly Ser 
    1475                 1480                 1485             


Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu Ser Pro  Lys Lys Lys 
    1490                 1495                 1500             


Arg Lys  Val 
    1505     


<210>  387
<211>  1513
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNHdelta with longer linker, 
       SV40+bipartiteSV40 NLS (TG14316)

<400>  387

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1505                 1510             


<210>  388
<211>  1616
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNH replaced with SSB, 
       SV40+bipartiteSV40 NLS (TG14317)

<400>  388

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Thr Thr  Glu Thr Phe Val Lys  Asp Ile Lys Pro Gly  Leu Lys Asn 
    1025                 1030                 1035             


Leu Asn  Leu Ile Phe Ile Val  Leu Glu Thr Gly Arg  Val Thr Lys 
    1040                 1045                 1050             


Thr Lys  Asp Gly His Glu Val  Arg Thr Cys Lys Val  Ala Asp Lys 
    1055                 1060                 1065             


Thr Gly  Ser Ile Asn Ile Ser  Val Trp Asp Asp Val  Gly Asn Leu 
    1070                 1075                 1080             


Ile Gln  Pro Gly Asp Ile Ile  Arg Leu Thr Lys Gly  Tyr Ala Ser 
    1085                 1090                 1095             


Val Phe  Lys Gly Cys Leu Thr  Leu Tyr Thr Gly Arg  Gly Gly Asp 
    1100                 1105                 1110             


Leu Gln  Lys Ile Gly Glu Phe  Cys Met Val Tyr Ser  Glu Val Pro 
    1115                 1120                 1125             


Asn Phe  Ser Glu Pro Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly 
    1130                 1135                 1140             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1145                 1150                 1155             


Val Ala  Arg Ile Leu Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys 
    1160                 1165                 1170             


Asn Asp  Lys Pro Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1175                 1180                 1185             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val 
    1190                 1195                 1200             


Arg Asp  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1205                 1210                 1215             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1220                 1225                 1230             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1235                 1240                 1245             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1250                 1255                 1260             


Arg Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val 
    1265                 1270                 1275             


Lys Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1280                 1285                 1290             


Asn Gly  Glu Thr Gly Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe 
    1295                 1300                 1305             


Ala Thr  Val Arg Lys Val Leu  Ala Met Pro Gln Val  Asn Ile Val 
    1310                 1315                 1320             


Lys Lys  Thr Glu Val Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile 
    1325                 1330                 1335             


Leu Ser  Lys Arg Glu Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly 
    1340                 1345                 1350             


Trp Asp  Thr Arg Lys Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala 
    1355                 1360                 1365             


Tyr Ser  Ile Leu Val Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys 
    1370                 1375                 1380             


Lys Leu  Lys Ser Val Lys Val  Leu Val Gly Ile Thr  Ile Met Glu 
    1385                 1390                 1395             


Lys Gly  Ser Tyr Glu Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys 
    1400                 1405                 1410             


Gly Tyr  Lys Asp Ile Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys 
    1415                 1420                 1425             


Tyr Ser  Leu Phe Glu Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala 
    1430                 1435                 1440             


Ser Ala  Thr Glu Leu Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln 
    1445                 1450                 1455             


His Leu  Val Arg Leu Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr 
    1460                 1465                 1470             


Thr Gly  Ser Asn Asn Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu 
    1475                 1480                 1485             


Phe Lys  Glu Ile Phe Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr 
    1490                 1495                 1500             


Ile Leu  Lys Asn Lys Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp 
    1505                 1510                 1515             


Glu Gln  Phe Ala Val Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe 
    1520                 1525                 1530             


Val Ser  Leu Leu Lys Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe 
    1535                 1540                 1545             


Thr Phe  Leu Asp Leu Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln 
    1550                 1555                 1560             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1565                 1570                 1575             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1580                 1585                 1590             


Asp Ser  Gly Gly Ser Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu 
    1595                 1600                 1605             


Ser Pro  Lys Lys Lys Arg Lys  Val 
    1610                 1615     


<210>  389
<211>  1488
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNHdelta with longer deletion 
       SV40+bipartiteSV40 NLS (TG14318)

<400>  389

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Gly Gly Ser  Gly Gly Leu 
        995                 1000                 1005             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1010                 1015                 1020             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1025                 1030                 1035             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1040                 1045                 1050             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1055                 1060                 1065             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1070                 1075                 1080             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1085                 1090                 1095             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1100                 1105                 1110             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1115                 1120                 1125             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1130                 1135                 1140             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1145                 1150                 1155             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1160                 1165                 1170             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1175                 1180                 1185             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1190                 1195                 1200             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1205                 1210                 1215             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1220                 1225                 1230             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1235                 1240                 1245             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1250                 1255                 1260             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1265                 1270                 1275             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1280                 1285                 1290             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1295                 1300                 1305             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1310                 1315                 1320             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1325                 1330                 1335             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1340                 1345                 1350             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1355                 1360                 1365             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1370                 1375                 1380             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1385                 1390                 1395             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1400                 1405                 1410             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1415                 1420                 1425             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1430                 1435                 1440             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1445                 1450                 1455             


Asp Leu  Ser Gln Leu Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr 
    1460                 1465                 1470             


Ala Asp  Gly Ser Glu Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1475                 1480                 1485             


<210>  390
<211>  1513
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNHdelta with longer positively
       charged linker, SV40+bipartiteSV40 NLS (TG14319)

<400>  390

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Lys 
    1010                 1015                 1020             


Ala Arg  Gly Lys Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1505                 1510             


<210>  391
<211>  1506
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNHdelta with shorter 
       positively charged linker, SV40+bipartiteSV40 NLS (TG14320)

<400>  391

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Lys Ser 
    1010                 1015                 1020             


Lys Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Ser  Gly Gly Phe Thr Phe  Leu Asp Leu 
    1430                 1435                 1440             


Asp Val  Lys Gln Gly Arg Leu  Arg Tyr Gln Thr Val  Thr Glu Val 
    1445                 1450                 1455             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1460                 1465                 1470             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Ser  Gly Gly Ser 
    1475                 1480                 1485             


Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu Ser Pro  Lys Lys Lys 
    1490                 1495                 1500             


Arg Lys  Val 
    1505     


<210>  392
<211>  1580
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNH replaced with StkC, 
       SV40+bipartiteSV40 NLS (TG14321)

<400>  392

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ser Val  Ser Met Val His Ser  Glu Arg Leu Pro Leu  Leu Met Asn 
    1025                 1030                 1035             


Arg Asn  Gly Ile Arg Phe Gln  Ser Glu Val Ile Asp  Pro Asn Pro 
    1040                 1045                 1050             


Pro Lys  Lys Val Lys Trp Val  Leu Thr Gly Ser Gly  Gly Asn Met 
    1055                 1060                 1065             


Phe Thr  Ala Asn Val Arg Gly  Ser Asn Val Ser Ser  Leu Val Lys 
    1070                 1075                 1080             


His Thr  Trp Ser Glu Ala Phe  Val Ser Leu Ile Glu  Asp Arg Gly 
    1085                 1090                 1095             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1100                 1105                 1110             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1115                 1120                 1125             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1130                 1135                 1140             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1145                 1150                 1155             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1160                 1165                 1170             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1175                 1180                 1185             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1190                 1195                 1200             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1205                 1210                 1215             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1220                 1225                 1230             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1235                 1240                 1245             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1250                 1255                 1260             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1265                 1270                 1275             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1280                 1285                 1290             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1295                 1300                 1305             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1310                 1315                 1320             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1325                 1330                 1335             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1340                 1345                 1350             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1355                 1360                 1365             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1370                 1375                 1380             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1385                 1390                 1395             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1400                 1405                 1410             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1415                 1420                 1425             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1430                 1435                 1440             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1445                 1450                 1455             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1460                 1465                 1470             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1475                 1480                 1485             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1490                 1495                 1500             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1505                 1510                 1515             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1520                 1525                 1530             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1535                 1540                 1545             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1550                 1555                 1560             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1565                 1570                 1575             


Lys Val  
    1580 


<210>  393
<211>  1499
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNHdelta with HNS linker, 
       SV40+bipartiteSV40 NLS (TG14322)

<400>  393

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Ala Ala Val  Lys Ser Gly 
        995                 1000                 1005             


Thr Lys  Ala Lys Arg Ala Gln  Arg Pro Gly Leu Ser  Glu Ala Asp 
    1010                 1015                 1020             


Lys Ala  Gly Phe Ile Lys Arg  Gln Leu Val Glu Thr  Arg Gln Ile 
    1025                 1030                 1035             


Thr Lys  His Val Ala Arg Ile  Leu Asp Ser Arg Met  Asn Thr Lys 
    1040                 1045                 1050             


Arg Asp  Lys Asn Asp Lys Pro  Ile Arg Glu Val Lys  Val Ile Thr 
    1055                 1060                 1065             


Leu Lys  Ser Lys Leu Val Ser  Asp Phe Arg Lys Asp  Phe Gln Leu 
    1070                 1075                 1080             


Tyr Lys  Val Arg Asp Ile Asn  Asn Tyr His His Ala  His Asp Ala 
    1085                 1090                 1095             


Tyr Leu  Asn Ala Val Val Gly  Thr Ala Leu Ile Lys  Lys Tyr Pro 
    1100                 1105                 1110             


Lys Leu  Glu Ser Glu Phe Val  Tyr Gly Asp Tyr Lys  Val Tyr Asp 
    1115                 1120                 1125             


Val Arg  Lys Met Ile Ala Lys  Ser Glu Gln Glu Ile  Gly Lys Ala 
    1130                 1135                 1140             


Thr Ala  Lys Arg Phe Phe Tyr  Ser Asn Ile Met Asn  Phe Phe Lys 
    1145                 1150                 1155             


Thr Glu  Val Lys Leu Ala Asn  Gly Glu Ile Arg Lys  Arg Pro Leu 
    1160                 1165                 1170             


Ile Glu  Thr Asn Gly Glu Thr  Gly Glu Val Val Trp  Asn Lys Glu 
    1175                 1180                 1185             


Lys Asp  Phe Ala Thr Val Arg  Lys Val Leu Ala Met  Pro Gln Val 
    1190                 1195                 1200             


Asn Ile  Val Lys Lys Thr Glu  Val Gln Thr Gly Gly  Phe Ser Lys 
    1205                 1210                 1215             


Glu Ser  Ile Leu Ser Lys Arg  Glu Ser Ala Lys Leu  Ile Pro Arg 
    1220                 1225                 1230             


Lys Lys  Gly Trp Asp Thr Arg  Lys Tyr Gly Gly Phe  Gly Ser Pro 
    1235                 1240                 1245             


Thr Val  Ala Tyr Ser Ile Leu  Val Val Ala Lys Val  Glu Lys Gly 
    1250                 1255                 1260             


Lys Ala  Lys Lys Leu Lys Ser  Val Lys Val Leu Val  Gly Ile Thr 
    1265                 1270                 1275             


Ile Met  Glu Lys Gly Ser Tyr  Glu Lys Asp Pro Ile  Gly Phe Leu 
    1280                 1285                 1290             


Glu Ala  Lys Gly Tyr Lys Asp  Ile Lys Lys Glu Leu  Ile Phe Lys 
    1295                 1300                 1305             


Leu Pro  Lys Tyr Ser Leu Phe  Glu Leu Glu Asn Gly  Arg Arg Arg 
    1310                 1315                 1320             


Met Leu  Ala Ser Ala Thr Glu  Leu Gln Lys Ala Asn  Glu Leu Val 
    1325                 1330                 1335             


Leu Pro  Gln His Leu Val Arg  Leu Leu Tyr Tyr Thr  Gln Asn Ile 
    1340                 1345                 1350             


Ser Ala  Thr Thr Gly Ser Asn  Asn Leu Gly Tyr Ile  Glu Gln His 
    1355                 1360                 1365             


Arg Glu  Glu Phe Lys Glu Ile  Phe Glu Lys Ile Ile  Asp Phe Ser 
    1370                 1375                 1380             


Glu Lys  Tyr Ile Leu Lys Asn  Lys Val Asn Ser Asn  Leu Lys Ser 
    1385                 1390                 1395             


Ser Phe  Asp Glu Gln Phe Ala  Val Ser Asp Ser Ile  Leu Leu Ser 
    1400                 1405                 1410             


Asn Ser  Phe Val Ser Leu Leu  Lys Tyr Thr Ser Phe  Gly Ala Ser 
    1415                 1420                 1425             


Gly Gly  Phe Thr Phe Leu Asp  Leu Asp Val Lys Gln  Gly Arg Leu 
    1430                 1435                 1440             


Arg Tyr  Gln Thr Val Thr Glu  Val Leu Asp Ala Thr  Leu Ile Tyr 
    1445                 1450                 1455             


Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg Thr Asp  Leu Ser Gln 
    1460                 1465                 1470             


Leu Gly  Gly Asp Ser Gly Gly  Ser Lys Arg Thr Ala  Asp Gly Ser 
    1475                 1480                 1485             


Glu Phe  Glu Ser Pro Lys Lys  Lys Arg Lys Val 
    1490                 1495                 


<210>  394
<211>  1609
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, ScLoop delta, 
       SV40+nucleoplasminNLS (TG14483)

<400>  394

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Ile Glu Glu Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu 
    1010                 1015                 1020             


Lys Glu  Asn Pro Val Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu 
    1025                 1030                 1035             


Tyr Leu  Tyr Tyr Leu Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln 
    1040                 1045                 1050             


Glu Leu  Asp Ile Asn Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile 
    1055                 1060                 1065             


Val Pro  Gln Ser Phe Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val 
    1070                 1075                 1080             


Leu Thr  Arg Ser Val Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro 
    1085                 1090                 1095             


Ser Glu  Glu Val Val Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu 
    1100                 1105                 1110             


Leu Asn  Ala Lys Leu Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr 
    1115                 1120                 1125             


Lys Ala  Glu Arg Gly Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe 
    1130                 1135                 1140             


Ile Lys  Arg Gln Leu Val Glu  Thr Arg Gln Ile Thr  Lys His Val 
    1145                 1150                 1155             


Ala Arg  Ile Leu Asp Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn 
    1160                 1165                 1170             


Asp Lys  Pro Ile Arg Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys 
    1175                 1180                 1185             


Leu Val  Ser Asp Phe Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg 
    1190                 1195                 1200             


Asp Ile  Asn Asn Tyr His His  Ala His Asp Ala Tyr  Leu Asn Ala 
    1205                 1210                 1215             


Val Val  Gly Thr Ala Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser 
    1220                 1225                 1230             


Glu Phe  Val Tyr Gly Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met 
    1235                 1240                 1245             


Ile Ala  Lys Ser Glu Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg 
    1250                 1255                 1260             


Phe Phe  Tyr Ser Asn Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys 
    1265                 1270                 1275             


Leu Ala  Asn Gly Glu Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn 
    1280                 1285                 1290             


Gly Glu  Thr Gly Glu Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala 
    1295                 1300                 1305             


Thr Val  Arg Lys Val Leu Ala  Met Pro Gln Val Asn  Ile Val Lys 
    1310                 1315                 1320             


Lys Thr  Glu Val Gln Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu 
    1325                 1330                 1335             


Ser Lys  Arg Glu Ser Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp 
    1340                 1345                 1350             


Asp Thr  Arg Lys Tyr Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr 
    1355                 1360                 1365             


Ser Ile  Leu Val Val Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys 
    1370                 1375                 1380             


Leu Lys  Ser Val Lys Val Leu  Val Gly Ile Thr Ile  Met Glu Lys 
    1385                 1390                 1395             


Gly Ser  Tyr Glu Lys Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly 
    1400                 1405                 1410             


Tyr Lys  Asp Ile Lys Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr 
    1415                 1420                 1425             


Ser Leu  Phe Glu Leu Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser 
    1430                 1435                 1440             


Ala Thr  Glu Leu Gln Lys Ala  Asn Glu Leu Val Leu  Pro Gln His 
    1445                 1450                 1455             


Leu Val  Arg Leu Leu Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr 
    1460                 1465                 1470             


Gly Ser  Asn Asn Leu Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe 
    1475                 1480                 1485             


Lys Glu  Ile Phe Glu Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile 
    1490                 1495                 1500             


Leu Lys  Asn Lys Val Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu 
    1505                 1510                 1515             


Gln Phe  Ala Val Ser Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val 
    1520                 1525                 1530             


Ser Leu  Leu Lys Tyr Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr 
    1535                 1540                 1545             


Phe Leu  Asp Leu Asp Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr 
    1550                 1555                 1560             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1565                 1570                 1575             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1580                 1585                 1590             


Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys 
    1595                 1600                 1605             


Lys 
    


<210>  395
<211>  1619
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, ScLoopQQ, SV40+nucleoplasminNLS
       (TG14484)

<400>  395

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala 
    1595                 1600                 1605             


Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys Lys 
    1610                 1615                 


<210>  396
<211>  1619
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, ScLoopAA, SV40+nucleoplasminNLS
       (TG14485)

<400>  396

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala 
    1595                 1600                 1605             


Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys Lys 
    1610                 1615                 


<210>  397
<211>  10629
<212>  DNA
<213>  Mus musculus


<220>
<221>  misc_feature
<223>  MPO chromosome 11, GRCm38.p4 C57BL/6J DNA SEQ. ACCESSION: 
       NC_000077

<400>  397
ataacttctc agctttagct ccctgaattt tggttctttc aaaggatttg ggacactcag       60

tttaaccaac actttgaggg gctggttgtt tgtgctctcc gggagaagct ctgatccatt      120

cctttgtcca cccctccttc ttcatagacc cctttctgtg cacccctaac cccagtgggg      180

gaggagagtc tagtcttggg ataggtccat ctctttttag gtcagcccgc ccccagcttt      240

tacaccccag gcataaaaac ccgttgtcag tgagaggagt tgacagtgtc aggtaagtgg      300

tgaagttgag ctcttgggga ctggggacta tagctggagg gggtcttggg ttgtacacct      360

cttcatagac caggcccact agggtagggc tgtgtataaa ggggagagga gggggcttgg      420

gaggaggaga attgtaaaac ctctcttttt tcaaaagctc tggatacttc ctgagctgag      480

gtttttgttt tttgtcttgt tttgttttgt tttgttttgt tttgttttgt tttgttttgt      540

tttgttttgt ttttttaaag gtttatttat ttattatatg taagtacact gcagctgtcc      600

ttagatactc cagaagagag catcagattt tgttacggat ggttgtgagc caccatgtgg      660

ttgctgggat ttgaactcgg gacctttgga agagcagtcg gcgctcttaa ccactgagca      720

atctccccag cccccttgtt ttttgtcttg cagagatgaa gctactcttg gccttggcag      780

gcctcctggc ccctctggcc atgcttcaga cctccaatgg tgccacccca ggtgatagtg      840

ccaaaaggag ggtggactaa gggattgggg gtagggggtg gggattctca gcccagcatc      900

tccccttccc ctggataggc ttcctttgct ttggggcctg atgtagggct ttggggccag      960

ccctgacctt atgatttatg ctctgggcag ctctcctggg ggaagtggag aattcagttg     1020

tgctgagctg tatggaagag gccaagcagc tagtggacag agcctacaag gagcggagag     1080

aaaggtgagg gtgccaggcc ctgtgtggcc ctgggcaaaa ctgcttcaag tcttctggag     1140

aaagggcaca cagtgggggg aggggggttg tagaaaggat gagggcctac aggtgaaatc     1200

gtacctggtc tgcctctact tgagtttctg gctgtcagaa catcttcagt ggccttgctt     1260

tctgtctgga catccaggtc catgtccttt tgacaaaaga aatgcactgc tcctccgtgg     1320

ttctctccag agttctttct gcctgctcac cgcaggctgg cctttgctga gtgtctctat     1380

cccccttctc tggccctccc ttgtctaaac agcatcaagc ggagcctcca aagcggctct     1440

gccagcccca cggagctcct gttttacttc aagcagccgg tggcgggcac cagaacagct     1500

gtgagggccg ctgattatct acatgtggcc ctagacctgc tgaagaggaa gctgcagccc     1560

ctgtggccaa ggcctttcaa tgttacaggt actctaactc cttttgtgct accacctcca     1620

ccccaagcaa cttctgtctc caagcctcca ttcaaggcag tggttcttaa cctgtgggtc     1680

gctacccctt cgggtacgtc aagtgaccct ctcacatggg tccacctaag accataggaa     1740

aacacaggtc ttaacattat gattcatctc agtagcaaaa ttacagttat gaagtagcaa     1800

gtaattttat agttctgttc accccatgag aaactggctt aaggggtctc tcagcattag     1860

caaacttgag aaccactggt ctaaggagtc tcggggcctc tgagctggcc cacccccctc     1920

ctcaccctca ctgggctact cagccccagg ttgcctagca taggaaatga ggtgggggaa     1980

ggctcggatc taagatcatg gctcactctt gcagatgtgt tgacacctgc tcagctgaat     2040

ctgttgtccg tgtcaagtgg ctgtgcctat caggacgtga gggtgacatg cccaccgaat     2100

gacaagtatc gcaccatcac tggacactgc aacaacaggt gtgggcccag ggcagttcag     2160

ggttgtgtgg tgtagggggt ctaggtcttg cctgtggcga cccattgggc cagagctctg     2220

tgcttccctc tagacgaagc cccactctgg gggcctccaa ccgtgccttt gtacgctggt     2280

tgcctgcaga gtatgaagat ggcgtctcca tgccctttgg ctggacgcct ggagtcaatc     2340

gcaatggctt caaggtgccc ctggtgagtg tccagtagag ggaggtgagc ccagctaagc     2400

ttatgtgcag gacctaggca tccaatgact cctgttgttg ctgcaggctc gccaggtctc     2460

caatgccatc gtgcgcttcc ccaacgatca gctgaccaag gaccaggagc gtgcactcat     2520

gttcatgcag tggggacagt ttctggatca tgatatcacc ttgactccag agccagctac     2580

ccggttctcc ttcttcactg gcctcaactg cgagaccagc tgcctgcagc agccaccctg     2640

cttccccctc aaggtgctgc ctccttccat ccatcagggt ctgtgtctga agagagggaa     2700

gatggtttac attagtgacg gcgccccttc tgagtcctag gtcctctcta gcccctgagc     2760

aagactggtc ctcgctcctg ggcccagagt caatacttgg ctccctcagc agagccagct     2820

gtcaactcaa ggaacctgat atgcaccctg atatgcaccc tgaacttagc atcgctctgt     2880

gtggtggggt gactggttct ttgtgaggct gggagaattc tgggatgagg atctgacttg     2940

tcttcttcat actcccaatg tacaggcact ggcctgttct cactctctct cccctatggg     3000

cgctccccta accctgcttc acagtgtgtg tgtgtgtgtg tgtgtgtgtg taaccctgct     3060

tcacagtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg     3120

tgtgtttggg ttgccccata gtcagggtca gggttgaagc accttctgtg taggggggaa     3180

agtgtggaga aaaagaaagg cagagattaa ggagataaag ggccaggggt ctgacagggc     3240

cagggtgaca gaacagagag gaaaaggcag acacaggcaa gaagaggatt aagaggcata     3300

gtgaagaggg gtccaagtgg ccttgcccaa ccccagaagg ctctctccat tccagttcag     3360

tgaattcttt gtcaccctta aacctcagtc tctttgctga ctgagcccca gtatttctat     3420

gaaaccttga gagcttccct gcagctcgag ggggccagag ttctcttctg cagggtgtaa     3480

gttacaaatg cccgctctcc cgacccacaa aagccagcct agcttctctc tgtgctgtag     3540

atcccaccca atgaccctcg aatcaagaac caaaaggact gcatcccctt cttccgctcc     3600

tgcccggcat gcaccaggaa caacatcacc attcgcaacc agatcaacgc gctcacttcc     3660

ttcgtggacg ccagcggggt gtacggcagc gaggaccccc tagccagaaa gctgcgcaac     3720

ctcaccaacc agctggggct gctggctatc aatacacgct tccaagacaa tggcagggcc     3780

ctgatgccct ttgacagcct gcacgatgac ccctgcctcc tcaccaaccg ctccgcccgc     3840

attccttgtt ttctggcagg tcagcccggg gactgaacta gagtggcata gagctgtttc     3900

cagcttgtga gaacacagcc tgtatgtgag caacatgcgg gcttgtgata agagatgctg     3960

gtctcaacaa cttattagtc tagttgcctt ggtaacgagt tggatggaga cagaaaggaa     4020

aataacagca aaaatcaaac cacataagca tacaaaaaaa aaatcctcca tccatcaact     4080

gaagatggag tcacagggct gctaaggacc ctctattatt gatccagctt tctctgtctt     4140

ctccctcctg tggactgtgg aaaaagagag agccaagtct atgtccacaa gggacagaga     4200

actaaaagtg acagagagga atatggcatc ctcacctttt agaaccatcc tgtccaatct     4260

ggagctagta gccacatgtg gttatttcaa tttaaattct ttgtagcagg gagtactggc     4320

tcacaccttt aatcccagtc tttgggaggt agaggcatga ggctcttagt gagaattcaa     4380

ggccagcctg gtctacagag tgagttctag gacagccagg gctatagagt gatagtgaga     4440

ccctgtcttg aaagaagaag aagaagaaga agaagaagaa gaagaagaag aagaagaaga     4500

agaagaagaa gaagaagaag aagaagaaga agaagaagaa gaagaagaaa acaaccaata     4560

aatacataaa taattttaaa ttaaattaaa agtttctgca gtaactgcat tttaagggct     4620

cagtagtcca tgggtgaaga agtggttgtt ttcttcacct cagaaaattc tagtagattg     4680

tgtcacagaa tatcactccg tgagcttttt ggtttcttag aaagacagaa ggtgggattt     4740

caaaaggaga gatgtttggc agtttgggga taggattggg gcaggggttg tggtcacacc     4800

cagaaattga tgcctctccc tcacacctct ctgtgatttt agactgggtt atagctagcc     4860

ttcgcaggtc tggtacttgc tttgaagccc aggctagctt caaacattcc tcaatcttcc     4920

tatcttagcc tgtcaagtgt tgggtggctc acatgtacca tcaagcccac acacagcggg     4980

aggtctcttc ttgcacctcc cttcccttct aggggtccag ttggtctctc ctgggtcccc     5040

tggcagggca gccaaggtca atgcaagggt gtgtctccta agggagggcc tgctctcttc     5100

tggactggtc atgacctctg gttactcatg gggtggaaaa gctactctca ggaatatgtt     5160

ttcctctcag aattttcttt ctaagttggc aagggctgag cccagtcagt gtcttgcagc     5220

aaaggccctg gcccccacct tctggccttc cctgttgaag gtttatggag cactagagga     5280

atcggggact tcattttctt gaatagtctt aggtatggat gtacacattc cgtgccctct     5340

tcccaccatg acaggatgta cagtggctta tagtgagagt gataatgaca actgtcttca     5400

aatagtagag ggtttggggt cctgttttga gaaggcttct gaggttgatc caatttgagc     5460

agaaaacatg catattactt tttggcttgt gccaatgagt acttgctctt gggagtgttc     5520

atgtattgtg gagtgtgtgt gtatgtgtgt gtagtatgta tgtatgtatg tatgcatgca     5580

tgcatataag tatgtgtagt atgtgtgtgt aatatgtctg tatgtctgtg tagtgtgtat     5640

gcatatgtat atatgtgtgt atgtatagta tgtgtatgta gtatgtgtat atgttattat     5700

gagtgtgtat agtatgtatg tgtatagtgt atgtgcatgc atatataatg tatatgtatg     5760

tgtgcatatg tgtgtagtat gtgtacatgt gtgtgtgtag taaatggcaa aatacagaaa     5820

gacacagggc ttggccttgt gcctaaggat cttgtgccta aggtctgctc caggcaggtg     5880

acaagtcaaa tgtaatggct attgggtaag gctgcaggga gtaccacagt gttcctgagc     5940

tcttatgcca gcctgtggct cacacttgaa tctggtgtgt gggagtaggt ggaaaacagg     6000

cacgtgaggg ccactgaagc tctttgctgg tcaggagatg atggagacca taaaggtgac     6060

aaaggctttg cttccctcag gggacatgcg ctccagcgag atgccggagc tcacctccat     6120

gcacaccctc tttgttcgag agcataaccg gctggccaca cagctcaagc gcctgaatcc     6180

tcgatggaat ggggagaagc tctaccagga ggcccggaag attgtagggg ccatggtcca     6240

ggtaggctgt gtcctaacac atcttggcca tgtgttcttg ggcagttttt ttttttaaat     6300

ctctcctgaa cactggcttt tttttatcca taaaaggctc aaaggagcga aatgaaaatg     6360

cccctcagac atgagatgct tggcgagtgc acttcccttc ctcctgtctc cccggccctc     6420

ttccttcctg agggcctgag gagttgagag ggtcttaagc agaagcacca gagtcaccgc     6480

tggctctttt acccgctgtc atcatcttga gcattccccc acccccaccc ccacccccac     6540

cccacccctg ccacccgggg cagggctcct ccatctggtt tttgatgttt tcatgacccc     6600

taatttccca agaatgtaaa cattcgtgca tatcacacac acacacacac acaacacaca     6660

cacacacaca cacacacaca tacacacact ccctagactt tagggagggg gtgtggttag     6720

tgctgagcat ggggttttga ttgacaggca cacccacaag agggattcac tttggtctca     6780

gaggcaattc tagccttagg tccagagtga cactgtctca cagcctttgg gggacatcgc     6840

actggagcag acctctgcca tctctctgtc tataaacctg tactgagccc tactgtgtgc     6900

tagtctattg ctggacactg tgaataatct cagtaatttc aacttccctg ggtctcaagg     6960

ggaaatggga ccactaatac cttaaagaca cgggggatat tacaatgact aaaaagacat     7020

tttgtttaat cctgagagag agggagagag aattattcct tgtttacatt tgtggttcag     7080

agagtttaat aaatgacctg aggtgagttc tatttatgtc cttctgttaa aatctaagac     7140

ctaagactat gggagactga tcctagccag gtaggggcaa ttcccccaag ctggagacca     7200

cagggaggaa caaatagaca tctaacacaa agtgactttg ttttagatca tcacataccg     7260

ggactacctg cccttggtgt tggggccagc agccatgaag aagtacctac cccagtaccg     7320

atcttacaac gactcagtag accctcgaat cgccaatgtc ttcaccaacg ctttccgtta     7380

tggccacacc ctcatccaac ccttcatgtt ccgcctgaac aatcagtacc ggcccacagg     7440

gcccaacccc cgagtcccac tcagcaaggt cttttttgcc agctggagag tcgtgttgga     7500

aggtgaatgg aacctaagaa aagccagcag ctaatgatgg gatggtggct gcagagacct     7560

aggctaggga gaggtctttt ttcatcttgg ggaacccagt cttgctttcc actttcattg     7620

acttagaagg ctagcgcatc ccaaccttgt gaaccctccc ccaaccctcc aagtgttgca     7680

agagcttata gcttctctct gacccctacg attccacttg gaccagccct gactccactg     7740

tgtgctctca tagtaagaga accaaccttt cctttcactc tggtctcttg tcctcaggtg     7800

gcattgaccc catcctccga ggcctcatgg ccactccagc caaactgaat cgccagaatc     7860

aaattgtggt ggatgagatc cgggagcgac tatttgagca agtcatgagg ataggactgg     7920

atttgcctgc tcttaacatg cagcgcagcc gggatcacgg cctcccaggt gagaggccag     7980

ccacagcccc atcccacctc ccaggcagct ccccttcttg ctcaggggtc ctttccagtt     8040

ttatggatgc acctgaggca aggcgctgcc ttttccagcc cctggtggcc taagttcggc     8100

ctctcttggc ttcttcacaa gctggacttt tgcaacccgc gtagcagctg aacatcatcc     8160

cacgatctgg aagtctgcct ccatcgttag cacgttctga ctcattcatg aactcgcggg     8220

tgggagagta ccactgtcat ttttactgac gaggaaacta gggcttagag cagttcagag     8280

tctggctcgg tgtggttgcc acgtcagtga gagacgtgga ggttggacct aaggggatca     8340

gcggaaccca aaagccccgt gcctgtgttg tggagtagca caagtggcca cgcctctggg     8400

aaacgttttc attctctgag tcagagaaag atacattagg ctgctgctct aagggacaat     8460

gtgctgtttc cgtgcttgag tctggcggga tttgtggtaa tacagggctg gccagggtag     8520

gagagagtga ggaaaagagt gagaggtctg gaggagccca gctgcctact ttggcctgag     8580

ccaatcattt agggatccat tctacttacg gtcttgtgta ctgagatggg cttttgaaca     8640

tctgtgtatt ctttcctgcc cttcccagta ctcctagccg gcaggagagg ggtccattta     8700

cagctacctg ttctctggcc caacgcagga tacaatgcct ggagacgctt ttgtgggctt     8760

ccacagccca gcacagtggg tgagctcggc acggtgctga agaacctgga gttggcacgg     8820

aagctgatgg cacaatatgg cacgcccaac aacattgaca tctggatggg cggtgtgtcc     8880

gagcccctgg agcccaatgg ccgtgtaggc cagctccttg cctgcctcat tggcactcag     8940

tttaggaagc tacgtgatgg tgatcggtaa ggaagctgtg ggaaagcagg gtgccacaac     9000

tgtgaggccg ggagggggcg catgcgcacc gcagcagtca ggcggcgctt agagaacaac     9060

ttatctgtga gttggttcca tcatgtctgg tctagggatg gtagtcaagt tctcagcctt     9120

ggtggtacgt gcctttgccc actgaaccat cttgctggtc ccaaaaatgc cactcttaat     9180

aggttctctg ggtttctcag tgctactgtg gaggcgtcag ttaccatgga aaagtgccaa     9240

aagtctcccg ctgacccctt ttcatgtcat ttcatcacga tcttggaatc tccgtgtcta     9300

gttcttatct agcttgcatt acacctttgg caatatcgta aaaaggtgtc ccagtcaaga     9360

agagaacatc acaaatgcca cccaggcttt cttttacccc gtgggagcgt gcatcagact     9420

gggacccaag cagctgtgct tctgtcttct gggctgagtg ggcactgccc acgacctgag     9480

ctctcctgtg ctgtagataa gaatattctg aagccccctc acactcaggg gacgtctcaa     9540

gccaggggat gtcctgggta gcctcactcg ggacatcctc agacataatg tccacgatgc     9600

tccctcccat gcaggttttg gtgggagaac ccaggcgtgt tcagtaaaca gcagagacag     9660

gccttggcca gcatctcctt gccccgtatc atctgtgaca acactggcat caccaccgtg     9720

tcgaagaaca acatcttcat gtccaacaca tacccccgag actttgtcag ctgtaacacc     9780

cttcctaaac tgaacctgac ttcctggaag gagacctaga ggttgggtaa gggcttggca     9840

cactgaaggg tatacctggt tggctagtgg gaatcacaga gaactctctt ccctgggtga     9900

atccatcttt gtggatattg tgggtgacag tggggggtgg ggggaaatga ctggacattc     9960

acttgtgagt gcacaagtat atttgcatat acacttgtgc acatctatat atatgcaaca    10020

tacataaact agtatttcat gtgtgtctgg tctgtgagca tgggcaatgt ttgatgtgcc    10080

ttgtgtgcct gccaatcaca tgagtctggg ctcactgctt acgacaatat taggaacgtg    10140

gggacagtga gcagacagcc cgagttcaag tcatcaccct gtagcacagt actgtgtgtg    10200

actttggcca aattactcta aggataaatt ctcttcctta taaatgaaat cagccacggc    10260

ttgttagagg gcctattgcc tcctccaact ctccactcat ggagcagctc ccatgtgcta    10320

ggcactgtgc tagccatggg aagcagaaac aataaagatg aataccaaag atgaatgcca    10380

aatcctcgat caccttctgg ttggcagaaa caaggtggtg gcctccggag gagaggatgc    10440

agctagcagc tgggctgtgt ggttgtgata cagttctggg ccagctctgc ttaactggct    10500

atttctcctt ttcccttact ctcttctctt gctattcaca ggagtctgcc gggagaagga    10560

cggaggccat tggccttcct gtgctatttg tacctgaggt tcacactaat aaaacatggt    10620

ggatgggga                                                            10629


<210>  398
<211>  718
<212>  PRT
<213>  Mus musculus


<220>
<221>  MISC_FEATURE
<223>  MPO PROTEIN SEQ. ACCESSION: NP_034954

<400>  398

Met Lys Leu Leu Leu Ala Leu Ala Gly Leu Leu Ala Pro Leu Ala Met 
1               5                   10                  15      


Leu Gln Thr Ser Asn Gly Ala Thr Pro Ala Leu Leu Gly Glu Val Glu 
            20                  25                  30          


Asn Ser Val Val Leu Ser Cys Met Glu Glu Ala Lys Gln Leu Val Asp 
        35                  40                  45              


Arg Ala Tyr Lys Glu Arg Arg Glu Ser Ile Lys Arg Ser Leu Gln Ser 
    50                  55                  60                  


Gly Ser Ala Ser Pro Thr Glu Leu Leu Phe Tyr Phe Lys Gln Pro Val 
65                  70                  75                  80  


Ala Gly Thr Arg Thr Ala Val Arg Ala Ala Asp Tyr Leu His Val Ala 
                85                  90                  95      


Leu Asp Leu Leu Lys Arg Lys Leu Gln Pro Leu Trp Pro Arg Pro Phe 
            100                 105                 110         


Asn Val Thr Asp Val Leu Thr Pro Ala Gln Leu Asn Leu Leu Ser Val 
        115                 120                 125             


Ser Ser Gly Cys Ala Tyr Gln Asp Val Arg Val Thr Cys Pro Pro Asn 
    130                 135                 140                 


Asp Lys Tyr Arg Thr Ile Thr Gly His Cys Asn Asn Arg Arg Ser Pro 
145                 150                 155                 160 


Thr Leu Gly Ala Ser Asn Arg Ala Phe Val Arg Trp Leu Pro Ala Glu 
                165                 170                 175     


Tyr Glu Asp Gly Val Ser Met Pro Phe Gly Trp Thr Pro Gly Val Asn 
            180                 185                 190         


Arg Asn Gly Phe Lys Val Pro Leu Ala Arg Gln Val Ser Asn Ala Ile 
        195                 200                 205             


Val Arg Phe Pro Asn Asp Gln Leu Thr Lys Asp Gln Glu Arg Ala Leu 
    210                 215                 220                 


Met Phe Met Gln Trp Gly Gln Phe Leu Asp His Asp Ile Thr Leu Thr 
225                 230                 235                 240 


Pro Glu Pro Ala Thr Arg Phe Ser Phe Phe Thr Gly Leu Asn Cys Glu 
                245                 250                 255     


Thr Ser Cys Leu Gln Gln Pro Pro Cys Phe Pro Leu Lys Ile Pro Pro 
            260                 265                 270         


Asn Asp Pro Arg Ile Lys Asn Gln Lys Asp Cys Ile Pro Phe Phe Arg 
        275                 280                 285             


Ser Cys Pro Ala Cys Thr Arg Asn Asn Ile Thr Ile Arg Asn Gln Ile 
    290                 295                 300                 


Asn Ala Leu Thr Ser Phe Val Asp Ala Ser Gly Val Tyr Gly Ser Glu 
305                 310                 315                 320 


Asp Pro Leu Ala Arg Lys Leu Arg Asn Leu Thr Asn Gln Leu Gly Leu 
                325                 330                 335     


Leu Ala Ile Asn Thr Arg Phe Gln Asp Asn Gly Arg Ala Leu Met Pro 
            340                 345                 350         


Phe Asp Ser Leu His Asp Asp Pro Cys Leu Leu Thr Asn Arg Ser Ala 
        355                 360                 365             


Arg Ile Pro Cys Phe Leu Ala Gly Asp Met Arg Ser Ser Glu Met Pro 
    370                 375                 380                 


Glu Leu Thr Ser Met His Thr Leu Phe Val Arg Glu His Asn Arg Leu 
385                 390                 395                 400 


Ala Thr Gln Leu Lys Arg Leu Asn Pro Arg Trp Asn Gly Glu Lys Leu 
                405                 410                 415     


Tyr Gln Glu Ala Arg Lys Ile Val Gly Ala Met Val Gln Ile Ile Thr 
            420                 425                 430         


Tyr Arg Asp Tyr Leu Pro Leu Val Leu Gly Pro Ala Ala Met Lys Lys 
        435                 440                 445             


Tyr Leu Pro Gln Tyr Arg Ser Tyr Asn Asp Ser Val Asp Pro Arg Ile 
    450                 455                 460                 


Ala Asn Val Phe Thr Asn Ala Phe Arg Tyr Gly His Thr Leu Ile Gln 
465                 470                 475                 480 


Pro Phe Met Phe Arg Leu Asn Asn Gln Tyr Arg Pro Thr Gly Pro Asn 
                485                 490                 495     


Pro Arg Val Pro Leu Ser Lys Val Phe Phe Ala Ser Trp Arg Val Val 
            500                 505                 510         


Leu Glu Gly Gly Ile Asp Pro Ile Leu Arg Gly Leu Met Ala Thr Pro 
        515                 520                 525             


Ala Lys Leu Asn Arg Gln Asn Gln Ile Val Val Asp Glu Ile Arg Glu 
    530                 535                 540                 


Arg Leu Phe Glu Gln Val Met Arg Ile Gly Leu Asp Leu Pro Ala Leu 
545                 550                 555                 560 


Asn Met Gln Arg Ser Arg Asp His Gly Leu Pro Gly Tyr Asn Ala Trp 
                565                 570                 575     


Arg Arg Phe Cys Gly Leu Pro Gln Pro Ser Thr Val Gly Glu Leu Gly 
            580                 585                 590         


Thr Val Leu Lys Asn Leu Glu Leu Ala Arg Lys Leu Met Ala Gln Tyr 
        595                 600                 605             


Gly Thr Pro Asn Asn Ile Asp Ile Trp Met Gly Gly Val Ser Glu Pro 
    610                 615                 620                 


Leu Glu Pro Asn Gly Arg Val Gly Gln Leu Leu Ala Cys Leu Ile Gly 
625                 630                 635                 640 


Thr Gln Phe Arg Lys Leu Arg Asp Gly Asp Arg Phe Trp Trp Glu Asn 
                645                 650                 655     


Pro Gly Val Phe Ser Lys Gln Gln Arg Gln Ala Leu Ala Ser Ile Ser 
            660                 665                 670         


Leu Pro Arg Ile Ile Cys Asp Asn Thr Gly Ile Thr Thr Val Ser Lys 
        675                 680                 685             


Asn Asn Ile Phe Met Ser Asn Thr Tyr Pro Arg Asp Phe Val Ser Cys 
    690                 695                 700                 


Asn Thr Leu Pro Lys Leu Asn Leu Thr Ser Trp Lys Glu Thr 
705                 710                 715             


<210>  399
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  short linker

<400>  399

Gly Gly Ser Gly 
1               


<210>  400
<211>  7
<212>  PRT
<213>  Simian virus 40


<220>
<221>  MISC_FEATURE
<223>  SV40 nuclear localization sequence

<400>  400

Pro Lys Lys Lys Arg Lys Val 
1               5           


<210>  401
<211>  16
<212>  PRT
<213>  Mus musculus


<220>
<221>  MISC_FEATURE
<223>  nucleoplasmin nuclear localization sequence

<400>  401

Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys 
1               5                   10                  15      


<210>  402
<211>  18
<212>  PRT
<213>  Simian virus 40


<220>
<221>  MISC_FEATURE
<223>  bipartite SV40 nuclear localization sequence

<400>  402

Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys Arg 
1               5                   10                  15      


Lys Val 
        


<210>  403
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  6 x His tag

<400>  403

His His His His His His 
1               5       


<210>  404
<211>  34
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TALE non-specific

<400>  404

Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ser Gly Gly Lys 
1               5                   10                  15      


Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala 
            20                  25                  30          


His Gly 
        


<210>  405
<211>  111
<212>  PRT
<213>  Homo sapiens


<220>
<221>  MISC_FEATURE
<223>  Human SSB1, domain 1

<400>  405

Met Thr Thr Glu Thr Phe Val Lys Asp Ile Lys Pro Gly Leu Lys Asn 
1               5                   10                  15      


Leu Asn Leu Ile Phe Ile Val Leu Glu Thr Gly Arg Val Thr Lys Thr 
            20                  25                  30          


Lys Asp Gly His Glu Val Arg Thr Cys Lys Val Ala Asp Lys Thr Gly 
        35                  40                  45              


Ser Ile Asn Ile Ser Val Trp Asp Asp Val Gly Asn Leu Ile Gln Pro 
    50                  55                  60                  


Gly Asp Ile Ile Arg Leu Thr Lys Gly Tyr Ala Ser Val Phe Lys Gly 
65                  70                  75                  80  


Cys Leu Thr Leu Tyr Thr Gly Arg Gly Gly Asp Leu Gln Lys Ile Gly 
                85                  90                  95      


Glu Phe Cys Met Val Tyr Ser Glu Val Pro Asn Phe Ser Glu Pro 
            100                 105                 110     


<210>  406
<211>  1526
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, HNH delta, AvrBS3 PAMBD loop replacement, 2NLS

<400>  406

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Gly  Gly Ser Ala Gly Leu  Thr Pro Glu 
    1430                 1435                 1440             


Gln Val  Val Ala Ile Ala Ser  Asn Ser Gly Gly Lys  Gln Ala Leu 
    1445                 1450                 1455             


Glu Thr  Val Gln Arg Leu Leu  Pro Val Leu Cys Gln  Ala His Gly 
    1460                 1465                 1470             


Gly Gly  Ser Gly Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1475                 1480                 1485             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1490                 1495                 1500             


Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys 
    1505                 1510                 1515             


Ala Gly  Gln Ala Lys Lys Lys  Lys 
    1520                 1525     


<210>  407
<211>  1512
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCas9-FokI, HNHdelta, AvrBS3 PAMBD domain replacement, 2NLS

<400>  407

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Gly Gly Ser Ala  Gly Gly Thr Gly Gly  Ser Leu Thr 
    1340                 1345                 1350             


Pro Glu  Gln Val Val Ala Ile  Ala Ser Asn Ser Gly  Gly Lys Gln 
    1355                 1360                 1365             


Ala Leu  Glu Thr Val Gln Arg  Leu Leu Pro Val Leu  Cys Gln Ala 
    1370                 1375                 1380             


His Gly  Leu Thr Pro Glu Gln  Val Val Ala Ile Ala  Ser Asn Ser 
    1385                 1390                 1395             


Gly Gly  Lys Gln Ala Leu Glu  Thr Val Gln Arg Leu  Leu Pro Val 
    1400                 1405                 1410             


Leu Cys  Gln Ala His Gly Leu  Thr Pro Glu Gln Val  Val Ala Ile 
    1415                 1420                 1425             


Ala Ser  Asn Ser Gly Gly Lys  Gln Ala Leu Glu Thr  Val Gln Arg 
    1430                 1435                 1440             


Leu Leu  Pro Val Leu Cys Gln  Ala His Gly Gly Gly  Ser Ala Gly 
    1445                 1450                 1455             


Gly Thr  Gly Gly Ser Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1460                 1465                 1470             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1475                 1480                 1485             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1490                 1495                 1500             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1505                 1510         


<210>  408
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon dScCasFok SCNA cassette 14082

<400>  408
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa ccaggcatgc agatcccaca gggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgtct gggcggtgct      600

acaactgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  409
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon dScCasFok SCNA cassette 14084

<400>  409
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa ccccagttgt agcaccgccc aggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacacccgcc tgtgggatct      600

gcatgcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  410
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon dScCasFok SCNA cassette 14086

<400>  410
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa cccagcccag ttgtagcacc gggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccaggg cgcctgtggg      600

atctgcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  411
<211>  389
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon Cas9 sgRNA cassette 14106

<400>  411
gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag       60

ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga      120

aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat      180

atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga      240

cgaaacaccg gcgccctggc cagtcgtctg ttttagagct agaaatagca agttaaaata      300

aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttacgcgtac      360

ttgcggccgc atattcctag gataccggt                                        389


<210>  412
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon dScCasFok SCNA cassette 14078

<400>  412
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa ctctggtggg gctgctccag gggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggcg ccctggccag      600

tcgtctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  413
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon dScCasFok SCNA cassette 14080

<400>  413
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa ctggtggggc tgctccaggc aggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacacccgcc ctggccagtc      600

gtctgggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  414
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon dScCasFok SCNA cassette 14088

<400>  414
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa catcccacag gcgccctggc cggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgcta caactgggct      600

ggcggcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  415
<211>  695
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon dScCasFok SCNA cassette 14090

<400>  415
acgcgtaaaa aaaaagcacc gactcggtgc cactttttca agttgataac ggactagcct       60

tattttaact tgctatttct agctctaaaa cacaggcgcc ctggccagtc gggtgtttcg      120

tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat      180

ttcccagaac acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag      240

ccatcttcct gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc      300

gattccttgg agcgggttga tgacgtcagc gttcaaagag ggcctatttc ccatgattcc      360

ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt      420

aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt      480

tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta      540

tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccaact gggctggcgg      600

ccaggagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga      660

aaaagtggca ccgagtcggt gcttttttta cgcgt                                 695


<210>  416
<211>  97
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RHO gene 5'-3' fragment of figure 6

<400>  416
cttctacgtg cccttctcca atgcgacggg tgtggtacgc agccacttcg agtacccaca       60

gtactacctg gctgagccat ggcagttctc catgctg                                97


<210>  417
<211>  98
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RHO gene 3'-5' fragment of figure 6

<400>  417
gaagatgcac gggaagaggt tacgctgccc acaccatgcg tcggtgaagc tcatgggtgt       60

catgatggac cgactcggta ccgtcaagag gtacgacc                               98


<210>  418
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  COMP gene 5'-3' fragment of figure 7

<400>  418
cccaggagga ctcagaccac gatggccagg gtgatgcctg cgacgacgac gacaatgacg       60

g                                                                       61


<210>  419
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  COMP gene 3'-5' fragment of figure 7

<400>  419
gggtcctcct gagtctggtg ctaccggtcc cactacggac gctgctgctg ctgttactgc       60

c                                                                       61


<210>  420
<211>  77
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PDCD1 gene 5'-3' fragment of figure 8

<400>  420
ccctgcgcct cctatgcgga gatgctactg ccactgctgc tgtcctcgct gctgggcggt       60

gagtgggcca agggccg                                                      77


<210>  421
<211>  77
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PDCD1 gene 3'-5' fragment of figure 8

<400>  421
gggacgcgga ggatacgcct ctacgatgac ggtgacgacg acaggagcga cgacccgcca       60

ctcacccggt tcccggc                                                      77


<210>  422
<211>  25
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PDCD1 amino acid sequence of figure 8

<400>  422

Met Gln Ile Pro Gln Ala Pro Trp Pro Val Val Trp Ala Val Leu Gln 
1               5                   10                  15      


Leu Gly Trp Arg Pro Gly Trp Phe Leu 
            20                  25  


<210>  423
<211>  15
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon drop-off probe TGEE40


<220>
<221>  misc_feature
<222>  (8)..(8)
<223>  g is a locked nucleic acid

<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  c is a locked nucleic acid

<400>  423
cgccctggcc agtcg                                                        15


<210>  424
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon forward primer

<400>  424
gtttcccttc cgctcac                                                      17


<210>  425
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PD-1 Exon reverse primer

<400>  425
gggactgaga gtgaaaggt                                                    19


<210>  426
<211>  1612
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, N-terminal HMGN, ancestral RuvC+Rec1/2, HNH delta with
       longer linker, SV40 and bipartite SV40 NLS

<400>  426

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val 
1               5                   10                  15      


Ser Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala 
            20                  25                  30          


Arg Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys 
        35                  40                  45              


Ala Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly 
    50                  55                  60                  


Lys Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr 
65                  70                  75                  80  


Lys Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser 
                85                  90                  95      


Pro Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly 
            100                 105                 110         


Ser Gly Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu 
        115                 120                 125             


Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile 
    130                 135                 140                 


Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val 
145                 150                 155                 160 


Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly 
                165                 170                 175     


Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile 
            180                 185                 190         


Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn 
        195                 200                 205             


Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn 
    210                 215                 220                 


Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr 
225                 230                 235                 240 


Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe 
                245                 250                 255     


Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn 
            260                 265                 270         


Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu 
        275                 280                 285             


Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe 
    290                 295                 300                 


Asn Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser 
305                 310                 315                 320 


Glu Ser Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu 
                325                 330                 335     


Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr 
            340                 345                 350         


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys 
        355                 360                 365             


Ser Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu 
    370                 375                 380                 


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
385                 390                 395                 400 


Arg Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu 
                405                 410                 415     


Met Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe 
            420                 425                 430         


Leu Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn 
        435                 440                 445             


Leu Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His 
    450                 455                 460                 


Leu Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu 
465                 470                 475                 480 


Ile Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu 
                485                 490                 495     


Ile Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe 
            500                 505                 510         


Tyr Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu 
        515                 520                 525             


Asp Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser 
    530                 535                 540                 


Lys Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys 
545                 550                 555                 560 


Lys Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr 
                565                 570                 575     


Pro Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln 
            580                 585                 590         


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln 
        595                 600                 605             


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser 
    610                 615                 620                 


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr 
625                 630                 635                 640 


Lys Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His 
                645                 650                 655     


Gln Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu 
            660                 665                 670         


Lys Tyr Lys Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
        675                 680                 685             


Tyr Ile Asp Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr 
    690                 695                 700                 


Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met 
705                 710                 715                 720 


Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu 
                725                 730                 735     


Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His 
            740                 745                 750         


Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro 
        755                 760                 765             


Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg 
    770                 775                 780                 


Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala 
785                 790                 795                 800 


Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu 
                805                 810                 815     


Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met 
            820                 825                 830         


Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His 
        835                 840                 845             


Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val 
    850                 855                 860                 


Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu 
865                 870                 875                 880 


Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val 
                885                 890                 895     


Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe 
            900                 905                 910         


Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu 
        915                 920                 925             


Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu 
    930                 935                 940                 


Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu 
945                 950                 955                 960 


Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr 
                965                 970                 975     


Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His 
            980                 985                 990         


Tyr Thr Gly Trp Gly Arg Leu Ser  Arg Lys Met Ile Asn  Gly Ile Arg 
        995                 1000                 1005             


Asp Lys  Gln Ser Gly Lys Thr  Ile Leu Asp Phe Leu  Lys Ser Asp 
    1010                 1015                 1020             


Gly Phe  Ser Asn Arg Asn Phe  Met Gln Leu Ile His  Asp Asp Ser 
    1025                 1030                 1035             


Leu Thr  Phe Lys Glu Glu Ile  Glu Lys Ala Gln Val  Ser Gly Gln 
    1040                 1045                 1050             


Gly Asp  Ser Leu His Glu Gln  Ile Ala Asp Leu Ala  Gly Ser Pro 
    1055                 1060                 1065             


Ala Ile  Lys Lys Gly Ile Leu  Gln Thr Val Lys Ile  Val Asp Glu 
    1070                 1075                 1080             


Leu Val  Lys Val Met Gly His  Lys Pro Glu Asn Ile  Val Ile Glu 
    1085                 1090                 1095             


Met Ala  Arg Glu Asn Gln Thr  Thr Thr Lys Gly Leu  Gln Gln Ser 
    1100                 1105                 1110             


Arg Glu  Arg Lys Lys Arg Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1115                 1120                 1125             


Ser Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1130                 1135                 1140             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1145                 1150                 1155             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1160                 1165                 1170             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1175                 1180                 1185             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1190                 1195                 1200             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1205                 1210                 1215             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1220                 1225                 1230             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1235                 1240                 1245             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1250                 1255                 1260             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1265                 1270                 1275             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1280                 1285                 1290             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1295                 1300                 1305             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1310                 1315                 1320             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1325                 1330                 1335             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1340                 1345                 1350             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1355                 1360                 1365             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1370                 1375                 1380             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1385                 1390                 1395             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1400                 1405                 1410             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1415                 1420                 1425             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1430                 1435                 1440             


Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro Gln His  Leu Val Arg 
    1445                 1450                 1455             


Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn 
    1460                 1465                 1470             


Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile 
    1475                 1480                 1485             


Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn 
    1490                 1495                 1500             


Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala 
    1505                 1510                 1515             


Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu 
    1520                 1525                 1530             


Lys Tyr  Thr Ser Phe Gly Ala  Ser Gly Gly Phe Thr  Phe Leu Asp 
    1535                 1540                 1545             


Leu Asp  Val Lys Gln Gly Arg  Leu Arg Tyr Gln Thr  Val Thr Glu 
    1550                 1555                 1560             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1565                 1570                 1575             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Ser Gly Gly 
    1580                 1585                 1590             


Ser Lys  Arg Thr Ala Asp Gly  Ser Glu Phe Glu Ser  Pro Lys Lys 
    1595                 1600                 1605             


Lys Arg  Lys Val 
    1610         


<210>  427
<211>  1541
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNH delta with longer linker, 
       PAMBD loop replaced with Zinc Finger+longer linker, SV40 and 
       bipartite SV40 NLS

<400>  427

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ala Gly 
    1430                 1435                 1440             


Gly Ser  Ala Gly Gly Thr Gly  Gly Ser Ala Gly Gly  Ser Gly Ala 
    1445                 1450                 1455             


Pro Tyr  Arg Cys Pro Cys Arg  Ala Gly Cys Pro Ser  Leu Ala Ser 
    1460                 1465                 1470             


Met Gln  Ala His Met Arg Gly  His Ser Gly Gly Ser  Gly Tyr Gln 
    1475                 1480                 1485             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1490                 1495                 1500             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1505                 1510                 1515             


Asp Ser  Gly Gly Ser Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu 
    1520                 1525                 1530             


Ser Pro  Lys Lys Lys Arg Lys  Val 
    1535                 1540     


<210>  428
<211>  1457
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNH delta with longer linker, 
       PAMBD replaced with Lac repressor DBD, SV40 and bipartite SV40 
       NLS

<400>  428

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Gly Met 
    1340                 1345                 1350             


Lys Pro  Val Thr Leu Tyr Asp  Val Ala Glu Tyr Ala  Gly Val Ser 
    1355                 1360                 1365             


Tyr Gln  Thr Val Ser Arg Val  Val Asn Gln Ala Ser  His Val Ser 
    1370                 1375                 1380             


Ala Lys  Thr Arg Glu Lys Val  Glu Ala Ala Met Ala  Glu Leu Asn 
    1385                 1390                 1395             


Gly Gly  Ser Gly Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1400                 1405                 1410             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1415                 1420                 1425             


Leu Ser  Gln Leu Gly Gly Asp  Ser Gly Gly Ser Lys  Arg Thr Ala 
    1430                 1435                 1440             


Asp Gly  Ser Glu Phe Glu Ser  Pro Lys Lys Lys Arg  Lys Val 
    1445                 1450                 1455         


<210>  429
<211>  1477
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, ancestral RuvC+Rec1/2, HNH delta with longer linker, 
       PAMBD replaced with Sto7, SV40 and bipartite SV40 NLS

<400>  429

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly 
    1340                 1345                 1350             


Met Ala  Thr Val Lys Phe Lys  Tyr Lys Gly Glu Glu  Lys Glu Val 
    1355                 1360                 1365             


Asp Ile  Ser Lys Ile Lys Lys  Val Trp Arg Val Gly  Lys Met Ile 
    1370                 1375                 1380             


Ser Phe  Thr Tyr Asp Glu Gly  Gly Gly Lys Thr Gly  Arg Gly Ala 
    1385                 1390                 1395             


Val Ser  Glu Lys Asp Ala Pro  Lys Glu Leu Leu Gln  Met Leu Glu 
    1400                 1405                 1410             


Lys Gln  Lys Lys Gly Gly Ser  Ala Gly Tyr Gln Thr  Val Thr Glu 
    1415                 1420                 1425             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1430                 1435                 1440             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Ser Gly Gly 
    1445                 1450                 1455             


Ser Lys  Arg Thr Ala Asp Gly  Ser Glu Phe Glu Ser  Pro Lys Lys 
    1460                 1465                 1470             


Lys Arg  Lys Val 
    1475         


<210>  430
<211>  1640
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, N-terminal HMGN, ancestral RuvC+Rec1/2, HNH delta with
       longer linker, PAMBD loop replaced with Zinc Finger+longer 
       linker, SV40 and bipartite SV40 NLS

<400>  430

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val 
1               5                   10                  15      


Ser Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala 
            20                  25                  30          


Arg Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys 
        35                  40                  45              


Ala Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly 
    50                  55                  60                  


Lys Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr 
65                  70                  75                  80  


Lys Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser 
                85                  90                  95      


Pro Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly 
            100                 105                 110         


Ser Gly Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu 
        115                 120                 125             


Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile 
    130                 135                 140                 


Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val 
145                 150                 155                 160 


Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly 
                165                 170                 175     


Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile 
            180                 185                 190         


Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn 
        195                 200                 205             


Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn 
    210                 215                 220                 


Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr 
225                 230                 235                 240 


Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe 
                245                 250                 255     


Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn 
            260                 265                 270         


Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu 
        275                 280                 285             


Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe 
    290                 295                 300                 


Asn Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser 
305                 310                 315                 320 


Glu Ser Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu 
                325                 330                 335     


Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr 
            340                 345                 350         


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys 
        355                 360                 365             


Ser Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu 
    370                 375                 380                 


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
385                 390                 395                 400 


Arg Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu 
                405                 410                 415     


Met Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe 
            420                 425                 430         


Leu Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn 
        435                 440                 445             


Leu Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His 
    450                 455                 460                 


Leu Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu 
465                 470                 475                 480 


Ile Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu 
                485                 490                 495     


Ile Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe 
            500                 505                 510         


Tyr Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu 
        515                 520                 525             


Asp Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser 
    530                 535                 540                 


Lys Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys 
545                 550                 555                 560 


Lys Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr 
                565                 570                 575     


Pro Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln 
            580                 585                 590         


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln 
        595                 600                 605             


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser 
    610                 615                 620                 


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr 
625                 630                 635                 640 


Lys Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His 
                645                 650                 655     


Gln Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu 
            660                 665                 670         


Lys Tyr Lys Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
        675                 680                 685             


Tyr Ile Asp Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr 
    690                 695                 700                 


Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met 
705                 710                 715                 720 


Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu 
                725                 730                 735     


Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His 
            740                 745                 750         


Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro 
        755                 760                 765             


Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg 
    770                 775                 780                 


Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala 
785                 790                 795                 800 


Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu 
                805                 810                 815     


Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met 
            820                 825                 830         


Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His 
        835                 840                 845             


Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val 
    850                 855                 860                 


Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu 
865                 870                 875                 880 


Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val 
                885                 890                 895     


Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe 
            900                 905                 910         


Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu 
        915                 920                 925             


Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu 
    930                 935                 940                 


Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu 
945                 950                 955                 960 


Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr 
                965                 970                 975     


Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His 
            980                 985                 990         


Tyr Thr Gly Trp Gly Arg Leu Ser  Arg Lys Met Ile Asn  Gly Ile Arg 
        995                 1000                 1005             


Asp Lys  Gln Ser Gly Lys Thr  Ile Leu Asp Phe Leu  Lys Ser Asp 
    1010                 1015                 1020             


Gly Phe  Ser Asn Arg Asn Phe  Met Gln Leu Ile His  Asp Asp Ser 
    1025                 1030                 1035             


Leu Thr  Phe Lys Glu Glu Ile  Glu Lys Ala Gln Val  Ser Gly Gln 
    1040                 1045                 1050             


Gly Asp  Ser Leu His Glu Gln  Ile Ala Asp Leu Ala  Gly Ser Pro 
    1055                 1060                 1065             


Ala Ile  Lys Lys Gly Ile Leu  Gln Thr Val Lys Ile  Val Asp Glu 
    1070                 1075                 1080             


Leu Val  Lys Val Met Gly His  Lys Pro Glu Asn Ile  Val Ile Glu 
    1085                 1090                 1095             


Met Ala  Arg Glu Asn Gln Thr  Thr Thr Lys Gly Leu  Gln Gln Ser 
    1100                 1105                 1110             


Arg Glu  Arg Lys Lys Arg Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1115                 1120                 1125             


Ser Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1130                 1135                 1140             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1145                 1150                 1155             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1160                 1165                 1170             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1175                 1180                 1185             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1190                 1195                 1200             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1205                 1210                 1215             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1220                 1225                 1230             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1235                 1240                 1245             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1250                 1255                 1260             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1265                 1270                 1275             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1280                 1285                 1290             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1295                 1300                 1305             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1310                 1315                 1320             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1325                 1330                 1335             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1340                 1345                 1350             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1355                 1360                 1365             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1370                 1375                 1380             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1385                 1390                 1395             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1400                 1405                 1410             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1415                 1420                 1425             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1430                 1435                 1440             


Leu Gln  Lys Ala Asn Glu Leu  Val Leu Pro Gln His  Leu Val Arg 
    1445                 1450                 1455             


Leu Leu  Tyr Tyr Thr Gln Asn  Ile Ser Ala Thr Thr  Gly Ser Asn 
    1460                 1465                 1470             


Asn Leu  Gly Tyr Ile Glu Gln  His Arg Glu Glu Phe  Lys Glu Ile 
    1475                 1480                 1485             


Phe Glu  Lys Ile Ile Asp Phe  Ser Glu Lys Tyr Ile  Leu Lys Asn 
    1490                 1495                 1500             


Lys Val  Asn Ser Asn Leu Lys  Ser Ser Phe Asp Glu  Gln Phe Ala 
    1505                 1510                 1515             


Val Ser  Asp Ser Ile Leu Leu  Ser Asn Ser Phe Val  Ser Leu Leu 
    1520                 1525                 1530             


Lys Tyr  Thr Ser Phe Gly Ala  Ala Gly Gly Ser Ala  Gly Gly Thr 
    1535                 1540                 1545             


Gly Gly  Ser Ala Gly Gly Ser  Gly Ala Pro Tyr Arg  Cys Pro Cys 
    1550                 1555                 1560             


Arg Ala  Gly Cys Pro Ser Leu  Ala Ser Met Gln Ala  His Met Arg 
    1565                 1570                 1575             


Gly His  Ser Gly Gly Ser Gly  Tyr Gln Thr Val Thr  Glu Val Leu 
    1580                 1585                 1590             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1595                 1600                 1605             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1610                 1615                 1620             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1625                 1630                 1635             


Lys Val  
    1640 


<210>  431
<211>  1556
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, N-terminal HMGN, ancestral RuvC+Rec1/2, HNH delta with
       longer linker, PAMBD replaced with Lac repressor DBD, SV40 and 
       bipartite SV40 NLS

<400>  431

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val 
1               5                   10                  15      


Ser Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala 
            20                  25                  30          


Arg Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys 
        35                  40                  45              


Ala Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly 
    50                  55                  60                  


Lys Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr 
65                  70                  75                  80  


Lys Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser 
                85                  90                  95      


Pro Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly 
            100                 105                 110         


Ser Gly Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu 
        115                 120                 125             


Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile 
    130                 135                 140                 


Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val 
145                 150                 155                 160 


Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly 
                165                 170                 175     


Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile 
            180                 185                 190         


Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn 
        195                 200                 205             


Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn 
    210                 215                 220                 


Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr 
225                 230                 235                 240 


Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe 
                245                 250                 255     


Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn 
            260                 265                 270         


Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu 
        275                 280                 285             


Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe 
    290                 295                 300                 


Asn Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser 
305                 310                 315                 320 


Glu Ser Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu 
                325                 330                 335     


Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr 
            340                 345                 350         


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys 
        355                 360                 365             


Ser Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu 
    370                 375                 380                 


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
385                 390                 395                 400 


Arg Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu 
                405                 410                 415     


Met Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe 
            420                 425                 430         


Leu Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn 
        435                 440                 445             


Leu Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His 
    450                 455                 460                 


Leu Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu 
465                 470                 475                 480 


Ile Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu 
                485                 490                 495     


Ile Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe 
            500                 505                 510         


Tyr Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu 
        515                 520                 525             


Asp Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser 
    530                 535                 540                 


Lys Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys 
545                 550                 555                 560 


Lys Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr 
                565                 570                 575     


Pro Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln 
            580                 585                 590         


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln 
        595                 600                 605             


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser 
    610                 615                 620                 


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr 
625                 630                 635                 640 


Lys Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His 
                645                 650                 655     


Gln Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu 
            660                 665                 670         


Lys Tyr Lys Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
        675                 680                 685             


Tyr Ile Asp Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr 
    690                 695                 700                 


Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met 
705                 710                 715                 720 


Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu 
                725                 730                 735     


Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His 
            740                 745                 750         


Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro 
        755                 760                 765             


Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg 
    770                 775                 780                 


Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala 
785                 790                 795                 800 


Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu 
                805                 810                 815     


Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met 
            820                 825                 830         


Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His 
        835                 840                 845             


Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val 
    850                 855                 860                 


Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu 
865                 870                 875                 880 


Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val 
                885                 890                 895     


Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe 
            900                 905                 910         


Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu 
        915                 920                 925             


Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu 
    930                 935                 940                 


Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu 
945                 950                 955                 960 


Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr 
                965                 970                 975     


Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His 
            980                 985                 990         


Tyr Thr Gly Trp Gly Arg Leu Ser  Arg Lys Met Ile Asn  Gly Ile Arg 
        995                 1000                 1005             


Asp Lys  Gln Ser Gly Lys Thr  Ile Leu Asp Phe Leu  Lys Ser Asp 
    1010                 1015                 1020             


Gly Phe  Ser Asn Arg Asn Phe  Met Gln Leu Ile His  Asp Asp Ser 
    1025                 1030                 1035             


Leu Thr  Phe Lys Glu Glu Ile  Glu Lys Ala Gln Val  Ser Gly Gln 
    1040                 1045                 1050             


Gly Asp  Ser Leu His Glu Gln  Ile Ala Asp Leu Ala  Gly Ser Pro 
    1055                 1060                 1065             


Ala Ile  Lys Lys Gly Ile Leu  Gln Thr Val Lys Ile  Val Asp Glu 
    1070                 1075                 1080             


Leu Val  Lys Val Met Gly His  Lys Pro Glu Asn Ile  Val Ile Glu 
    1085                 1090                 1095             


Met Ala  Arg Glu Asn Gln Thr  Thr Thr Lys Gly Leu  Gln Gln Ser 
    1100                 1105                 1110             


Arg Glu  Arg Lys Lys Arg Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1115                 1120                 1125             


Ser Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1130                 1135                 1140             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1145                 1150                 1155             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1160                 1165                 1170             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1175                 1180                 1185             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1190                 1195                 1200             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1205                 1210                 1215             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1220                 1225                 1230             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1235                 1240                 1245             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1250                 1255                 1260             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1265                 1270                 1275             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1280                 1285                 1290             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1295                 1300                 1305             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1310                 1315                 1320             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1325                 1330                 1335             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1340                 1345                 1350             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1355                 1360                 1365             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1370                 1375                 1380             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1385                 1390                 1395             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1400                 1405                 1410             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1415                 1420                 1425             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1430                 1435                 1440             


Leu Gln  Lys Ala Gly Gly Ser  Gly Met Lys Pro Val  Thr Leu Tyr 
    1445                 1450                 1455             


Asp Val  Ala Glu Tyr Ala Gly  Val Ser Tyr Gln Thr  Val Ser Arg 
    1460                 1465                 1470             


Val Val  Asn Gln Ala Ser His  Val Ser Ala Lys Thr  Arg Glu Lys 
    1475                 1480                 1485             


Val Glu  Ala Ala Met Ala Glu  Leu Asn Gly Gly Ser  Gly Tyr Gln 
    1490                 1495                 1500             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1505                 1510                 1515             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1520                 1525                 1530             


Asp Ser  Gly Gly Ser Lys Arg  Thr Ala Asp Gly Ser  Glu Phe Glu 
    1535                 1540                 1545             


Ser Pro  Lys Lys Lys Arg Lys  Val 
    1550                 1555     


<210>  432
<211>  1576
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, N-terminal HMGN, ancestral RuvC+Rec1/2, HNH delta with
       longer linker, PAMBD replaced with SSO7D, SV40 and bipartite SV40
       NLS

<400>  432

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Pro Lys Arg Lys Val 
1               5                   10                  15      


Ser Ser Ala Glu Gly Ala Ala Lys Glu Glu Pro Lys Arg Arg Ser Ala 
            20                  25                  30          


Arg Leu Ser Ala Lys Pro Pro Ala Lys Val Glu Ala Lys Pro Lys Lys 
        35                  40                  45              


Ala Ala Ala Lys Asp Lys Ser Ser Asp Lys Lys Val Gln Thr Lys Gly 
    50                  55                  60                  


Lys Arg Gly Ala Lys Gly Lys Gln Ala Glu Val Ala Asn Gln Glu Thr 
65                  70                  75                  80  


Lys Glu Asp Leu Pro Ala Glu Asn Gly Glu Thr Lys Thr Glu Glu Ser 
                85                  90                  95      


Pro Ala Ser Asp Glu Ala Gly Glu Lys Glu Ala Lys Ser Asp Gly Gly 
            100                 105                 110         


Ser Gly Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu 
        115                 120                 125             


Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile 
    130                 135                 140                 


Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val 
145                 150                 155                 160 


Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly 
                165                 170                 175     


Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile 
            180                 185                 190         


Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn 
        195                 200                 205             


Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn 
    210                 215                 220                 


Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr 
225                 230                 235                 240 


Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe 
                245                 250                 255     


Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn 
            260                 265                 270         


Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu 
        275                 280                 285             


Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe 
    290                 295                 300                 


Asn Asn Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser 
305                 310                 315                 320 


Glu Ser Ala Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu 
                325                 330                 335     


Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr 
            340                 345                 350         


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys 
        355                 360                 365             


Ser Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu 
    370                 375                 380                 


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
385                 390                 395                 400 


Arg Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu 
                405                 410                 415     


Met Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe 
            420                 425                 430         


Leu Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn 
        435                 440                 445             


Leu Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His 
    450                 455                 460                 


Leu Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu 
465                 470                 475                 480 


Ile Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu 
                485                 490                 495     


Ile Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe 
            500                 505                 510         


Tyr Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu 
        515                 520                 525             


Asp Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser 
    530                 535                 540                 


Lys Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys 
545                 550                 555                 560 


Lys Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr 
                565                 570                 575     


Pro Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln 
            580                 585                 590         


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln 
        595                 600                 605             


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser 
    610                 615                 620                 


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr 
625                 630                 635                 640 


Lys Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His 
                645                 650                 655     


Gln Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu 
            660                 665                 670         


Lys Tyr Lys Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
        675                 680                 685             


Tyr Ile Asp Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr 
    690                 695                 700                 


Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met 
705                 710                 715                 720 


Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu 
                725                 730                 735     


Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His 
            740                 745                 750         


Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro 
        755                 760                 765             


Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg 
    770                 775                 780                 


Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala 
785                 790                 795                 800 


Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu 
                805                 810                 815     


Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met 
            820                 825                 830         


Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His 
        835                 840                 845             


Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val 
    850                 855                 860                 


Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu 
865                 870                 875                 880 


Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val 
                885                 890                 895     


Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe 
            900                 905                 910         


Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu 
        915                 920                 925             


Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu 
    930                 935                 940                 


Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu 
945                 950                 955                 960 


Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr 
                965                 970                 975     


Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His 
            980                 985                 990         


Tyr Thr Gly Trp Gly Arg Leu Ser  Arg Lys Met Ile Asn  Gly Ile Arg 
        995                 1000                 1005             


Asp Lys  Gln Ser Gly Lys Thr  Ile Leu Asp Phe Leu  Lys Ser Asp 
    1010                 1015                 1020             


Gly Phe  Ser Asn Arg Asn Phe  Met Gln Leu Ile His  Asp Asp Ser 
    1025                 1030                 1035             


Leu Thr  Phe Lys Glu Glu Ile  Glu Lys Ala Gln Val  Ser Gly Gln 
    1040                 1045                 1050             


Gly Asp  Ser Leu His Glu Gln  Ile Ala Asp Leu Ala  Gly Ser Pro 
    1055                 1060                 1065             


Ala Ile  Lys Lys Gly Ile Leu  Gln Thr Val Lys Ile  Val Asp Glu 
    1070                 1075                 1080             


Leu Val  Lys Val Met Gly His  Lys Pro Glu Asn Ile  Val Ile Glu 
    1085                 1090                 1095             


Met Ala  Arg Glu Asn Gln Thr  Thr Thr Lys Gly Leu  Gln Gln Ser 
    1100                 1105                 1110             


Arg Glu  Arg Lys Lys Arg Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1115                 1120                 1125             


Ser Gly  Gly Leu Ser Glu Ala  Asp Lys Ala Gly Phe  Ile Lys Arg 
    1130                 1135                 1140             


Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys His Val  Ala Arg Ile 
    1145                 1150                 1155             


Leu Asp  Ser Arg Met Asn Thr  Lys Arg Asp Lys Asn  Asp Lys Pro 
    1160                 1165                 1170             


Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys Ser Lys  Leu Val Ser 
    1175                 1180                 1185             


Asp Phe  Arg Lys Asp Phe Gln  Leu Tyr Lys Val Arg  Asp Ile Asn 
    1190                 1195                 1200             


Asn Tyr  His His Ala His Asp  Ala Tyr Leu Asn Ala  Val Val Gly 
    1205                 1210                 1215             


Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
    1220                 1225                 1230             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1235                 1240                 1245             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Arg  Phe Phe Tyr 
    1250                 1255                 1260             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Val Lys  Leu Ala Asn 
    1265                 1270                 1275             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1280                 1285                 1290             


Gly Glu  Val Val Trp Asn Lys  Glu Lys Asp Phe Ala  Thr Val Arg 
    1295                 1300                 1305             


Lys Val  Leu Ala Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1310                 1315                 1320             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Ser Lys Arg 
    1325                 1330                 1335             


Glu Ser  Ala Lys Leu Ile Pro  Arg Lys Lys Gly Trp  Asp Thr Arg 
    1340                 1345                 1350             


Lys Tyr  Gly Gly Phe Gly Ser  Pro Thr Val Ala Tyr  Ser Ile Leu 
    1355                 1360                 1365             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ala Lys Lys  Leu Lys Ser 
    1370                 1375                 1380             


Val Lys  Val Leu Val Gly Ile  Thr Ile Met Glu Lys  Gly Ser Tyr 
    1385                 1390                 1395             


Glu Lys  Asp Pro Ile Gly Phe  Leu Glu Ala Lys Gly  Tyr Lys Asp 
    1400                 1405                 1410             


Ile Lys  Lys Glu Leu Ile Phe  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1415                 1420                 1425             


Glu Leu  Glu Asn Gly Arg Arg  Arg Met Leu Ala Ser  Ala Thr Glu 
    1430                 1435                 1440             


Leu Gln  Lys Ala Gly Gly Ser  Ala Gly Met Ala Thr  Val Lys Phe 
    1445                 1450                 1455             


Lys Tyr  Lys Gly Glu Glu Lys  Glu Val Asp Ile Ser  Lys Ile Lys 
    1460                 1465                 1470             


Lys Val  Trp Arg Val Gly Lys  Met Ile Ser Phe Thr  Tyr Asp Glu 
    1475                 1480                 1485             


Gly Gly  Gly Lys Thr Gly Arg  Gly Ala Val Ser Glu  Lys Asp Ala 
    1490                 1495                 1500             


Pro Lys  Glu Leu Leu Gln Met  Leu Glu Lys Gln Lys  Lys Gly Gly 
    1505                 1510                 1515             


Ser Ala  Gly Tyr Gln Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu 
    1520                 1525                 1530             


Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu 
    1535                 1540                 1545             


Ser Gln  Leu Gly Gly Asp Ser  Gly Gly Ser Lys Arg  Thr Ala Asp 
    1550                 1555                 1560             


Gly Ser  Glu Phe Glu Ser Pro  Lys Lys Lys Arg Lys  Val 
    1565                 1570                 1575     


<210>  433
<211>  1488
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok,HNH delta,PAM replace with Sto7,design 7, 
       SV40+nucleoplasmin NLS

<400>  433

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser 
    1340                 1345                 1350             


Ala Gly  Gly Val Thr Val Lys  Phe Lys Tyr Lys Gly  Glu Glu Lys 
    1355                 1360                 1365             


Glu Val  Asp Ile Ser Lys Ile  Lys Lys Val Trp Arg  Val Gly Lys 
    1370                 1375                 1380             


Met Ile  Ser Phe Thr Tyr Asp  Asp Asn Gly Lys Thr  Gly Arg Gly 
    1385                 1390                 1395             


Ala Val  Ser Glu Lys Asp Ala  Pro Lys Glu Leu Leu  Gln Met Leu 
    1400                 1405                 1410             


Glu Lys  Ser Gly Lys Lys Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1415                 1420                 1425             


Ser Ala  Gly Gly Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1430                 1435                 1440             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1445                 1450                 1455             


Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys 
    1460                 1465                 1470             


Ala Gly  Gln Ala Lys Lys Lys  Lys Pro Lys Lys Lys  Arg Lys Val 
    1475                 1480                 1485             


<210>  434
<211>  1575
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok,HNH delta,PAM replace with Sto7,design 8, 
       SV40+nucleoplasmin NLS

<400>  434

Met Ala Val Thr Ala Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly 
1               5                   10                  15      


Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg 
            20                  25                  30          


His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile 
        35                  40                  45              


Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu 
    50                  55                  60                  


Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser 
65                  70                  75                  80  


Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr 
                85                  90                  95      


Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro 
            100                 105                 110         


Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr 
        115                 120                 125             


Arg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser 
    130                 135                 140                 


Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly 
145                 150                 155                 160 


Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn 
                165                 170                 175     


Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile 
            180                 185                 190         


Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn 
        195                 200                 205             


Gly Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser 
    210                 215                 220                 


Ala Thr Pro Glu Ser Gly Thr Glu Lys Lys Tyr Ser Ile Gly Leu Ala 
225                 230                 235                 240 


Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys 
                245                 250                 255     


Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser 
            260                 265                 270         


Ile Lys Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr 
        275                 280                 285             


Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg 
    290                 295                 300                 


Arg Lys Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met 
305                 310                 315                 320 


Ala Lys Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu 
                325                 330                 335     


Val Glu Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu 
            340                 345                 350         


Ala Asp Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu 
        355                 360                 365             


Arg Lys Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile 
    370                 375                 380                 


Tyr Leu Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile 
385                 390                 395                 400 


Glu Gly Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr 
                405                 410                 415     


Gln Leu Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp 
            420                 425                 430         


Glu Ile Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys 
        435                 440                 445             


Ser Lys Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys 
    450                 455                 460                 


Asn Gly Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro 
465                 470                 475                 480 


Asn Phe Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu 
                485                 490                 495     


Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile 
            500                 505                 510         


Gly Asp Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp 
        515                 520                 525             


Ala Ile Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys 
    530                 535                 540                 


Ala Pro Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln 
545                 550                 555                 560 


Asp Leu Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys 
                565                 570                 575     


Tyr Ala Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr 
            580                 585                 590         


Val Gly Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr 
        595                 600                 605             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    610                 615                 620                 


Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg 
625                 630                 635                 640 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                645                 650                 655     


Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe 
            660                 665                 670         


Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        675                 680                 685             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    690                 695                 700                 


Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu 
705                 710                 715                 720 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                725                 730                 735     


Asn Phe Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser 
            740                 745                 750         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        755                 760                 765             


Tyr Val Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln 
    770                 775                 780                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
785                 790                 795                 800 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                805                 810                 815     


Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            820                 825                 830         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        835                 840                 845             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    850                 855                 860                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
865                 870                 875                 880 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr 
                885                 890                 895     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp 
            900                 905                 910         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        915                 920                 925             


Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    930                 935                 940                 


Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
945                 950                 955                 960 


His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                965                 970                 975     


Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly 
            980                 985                 990         


His Lys Pro Glu Asn Ile Val Ile  Glu Met Ala Arg Glu  Asn Gln Thr 
        995                 1000                 1005             


Thr Thr  Lys Gly Leu Gln Gln  Ser Arg Glu Arg Lys  Lys Arg Gly 
    1010                 1015                 1020             


Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys 
    1025                 1030                 1035             


Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr Lys His  Val Ala Arg 
    1040                 1045                 1050             


Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys 
    1055                 1060                 1065             


Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val 
    1070                 1075                 1080             


Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile 
    1085                 1090                 1095             


Asn Asn  Tyr His His Ala His  Asp Ala Tyr Leu Asn  Ala Val Val 
    1100                 1105                 1110             


Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
    1115                 1120                 1125             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1130                 1135                 1140             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe 
    1145                 1150                 1155             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala 
    1160                 1165                 1170             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1175                 1180                 1185             


Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val 
    1190                 1195                 1200             


Arg Lys  Val Leu Ala Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1205                 1210                 1215             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys 
    1220                 1225                 1230             


Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr 
    1235                 1240                 1245             


Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile 
    1250                 1255                 1260             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys 
    1265                 1270                 1275             


Ser Val  Lys Val Leu Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser 
    1280                 1285                 1290             


Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1295                 1300                 1305             


Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu 
    1310                 1315                 1320             


Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr 
    1325                 1330                 1335             


Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu Pro Gln  His Leu Val 
    1340                 1345                 1350             


Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser 
    1355                 1360                 1365             


Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu 
    1370                 1375                 1380             


Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys 
    1385                 1390                 1395             


Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe 
    1400                 1405                 1410             


Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu 
    1415                 1420                 1425             


Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly Ser Ala  Gly Gly Thr 
    1430                 1435                 1440             


Gly Gly  Ser Val Thr Val Lys  Phe Ala Tyr Ala Gly  Glu Glu Lys 
    1445                 1450                 1455             


Glu Val  Asp Ile Ser Lys Ile  Lys Ala Val Trp Arg  Val Gly Ala 
    1460                 1465                 1470             


Met Ile  Ser Phe Thr Tyr Asp  Asp Asn Gly Lys Thr  Gly Ala Gly 
    1475                 1480                 1485             


Ala Val  Ser Glu Lys Asp Ala  Pro Lys Glu Leu Leu  Gln Met Leu 
    1490                 1495                 1500             


Glu Lys  Ser Gly Lys Lys Gly  Gly Ser Ala Gly Gly  Thr Gly Gly 
    1505                 1510                 1515             


Ser Tyr  Gln Thr Val Thr Glu  Val Leu Asp Ala Thr  Leu Ile Tyr 
    1520                 1525                 1530             


Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg Thr Asp  Leu Ser Gln 
    1535                 1540                 1545             


Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr Lys Lys  Ala Gly Gln 
    1550                 1555                 1560             


Ala Lys  Lys Lys Lys Pro Lys  Lys Lys Arg Lys Val  
    1565                 1570                 1575 


<210>  435
<211>  1590
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok,HNH delta,PAM replace with HMGN,design 1, 
       SV40+nucleoplasmin NLS

<400>  435

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Gly  Gly Ser Gly Pro Lys  Arg Lys Val 
    1430                 1435                 1440             


Ser Ser  Ala Glu Gly Ala Ala  Lys Glu Glu Pro Lys  Arg Arg Ser 
    1445                 1450                 1455             


Ala Arg  Leu Ser Ala Lys Pro  Pro Ala Lys Val Glu  Ala Lys Pro 
    1460                 1465                 1470             


Lys Lys  Ala Ala Ala Lys Asp  Lys Ser Ser Asp Lys  Lys Val Gln 
    1475                 1480                 1485             


Thr Lys  Gly Lys Arg Gly Ala  Lys Gly Lys Gln Ala  Glu Val Ala 
    1490                 1495                 1500             


Asn Gln  Glu Thr Lys Glu Asp  Leu Pro Ala Glu Asn  Gly Glu Thr 
    1505                 1510                 1515             


Lys Thr  Glu Glu Ser Pro Ala  Ser Asp Glu Ala Gly  Glu Lys Glu 
    1520                 1525                 1530             


Ala Lys  Ser Asp Gly Gly Ser  Gly Tyr Gln Thr Val  Thr Glu Val 
    1535                 1540                 1545             


Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile Thr Gly  Leu Tyr Glu 
    1550                 1555                 1560             


Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly Asp Lys  Arg Pro Ala 
    1565                 1570                 1575             


Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys Lys Lys  
    1580                 1585                 1590 


<210>  436
<211>  1497
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok,HNH delta,PAM replace with HMGN,design 2, 
       SV40+nucleoplasmin NLS

<400>  436

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Gly Gly Ser Gly  Pro Lys Arg Lys Val  Ser Ser Ala 
    1340                 1345                 1350             


Glu Gly  Ala Ala Lys Glu Glu  Pro Lys Arg Arg Ser  Ala Arg Leu 
    1355                 1360                 1365             


Ser Ala  Lys Pro Pro Ala Lys  Val Glu Ala Lys Pro  Lys Lys Ala 
    1370                 1375                 1380             


Ala Ala  Lys Asp Lys Ser Ser  Asp Lys Lys Val Gln  Thr Lys Gly 
    1385                 1390                 1395             


Lys Arg  Gly Ala Lys Gly Lys  Gln Ala Glu Val Ala  Asn Gln Glu 
    1400                 1405                 1410             


Thr Lys  Glu Asp Leu Pro Ala  Glu Asn Gly Glu Thr  Lys Thr Glu 
    1415                 1420                 1425             


Glu Ser  Pro Ala Ser Asp Glu  Ala Gly Glu Lys Glu  Ala Lys Ser 
    1430                 1435                 1440             


Asp Gly  Gly Ser Gly Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1445                 1450                 1455             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1460                 1465                 1470             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1475                 1480                 1485             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1490                 1495         


<210>  437
<211>  1565
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok,HNHdelta,PAM replace with StkC,design 1, 
       SV40+nucleoplasmin NLS

<400>  437

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Asn Glu Leu Val  Leu Pro Gln His Leu  Val Arg Leu 
    1340                 1345                 1350             


Leu Tyr  Tyr Thr Gln Asn Ile  Ser Ala Thr Thr Gly  Ser Asn Asn 
    1355                 1360                 1365             


Leu Gly  Tyr Ile Glu Gln His  Arg Glu Glu Phe Lys  Glu Ile Phe 
    1370                 1375                 1380             


Glu Lys  Ile Ile Asp Phe Ser  Glu Lys Tyr Ile Leu  Lys Asn Lys 
    1385                 1390                 1395             


Val Asn  Ser Asn Leu Lys Ser  Ser Phe Asp Glu Gln  Phe Ala Val 
    1400                 1405                 1410             


Ser Asp  Ser Ile Leu Leu Ser  Asn Ser Phe Val Ser  Leu Leu Lys 
    1415                 1420                 1425             


Tyr Thr  Ser Phe Gly Ala Gly  Gly Ser Gly Ser Val  Ser Met Val 
    1430                 1435                 1440             


His Ser  Glu Arg Leu Pro Leu  Leu Met Asn Arg Asn  Gly Ile Arg 
    1445                 1450                 1455             


Phe Gln  Ser Glu Val Ile Asp  Pro Asn Pro Pro Lys  Lys Val Lys 
    1460                 1465                 1470             


Trp Val  Leu Thr Gly Ser Gly  Gly Asn Met Phe Thr  Ala Asn Val 
    1475                 1480                 1485             


Arg Gly  Ser Asn Val Ser Ser  Leu Val Lys His Thr  Trp Ser Glu 
    1490                 1495                 1500             


Ala Phe  Val Ser Leu Ile Glu  Asp Arg Gly Gly Ser  Gly Tyr Gln 
    1505                 1510                 1515             


Thr Val  Thr Glu Val Leu Asp  Ala Thr Leu Ile Tyr  Gln Ser Ile 
    1520                 1525                 1530             


Thr Gly  Leu Tyr Glu Thr Arg  Thr Asp Leu Ser Gln  Leu Gly Gly 
    1535                 1540                 1545             


Asp Lys  Arg Pro Ala Ala Thr  Lys Lys Ala Gly Gln  Ala Lys Lys 
    1550                 1555                 1560             


Lys Lys  
    1565 


<210>  438
<211>  1472
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok,HNHdelta,PAM replace with StkC,design 2, 
       SV40+nucleoplasmin NLS

<400>  438

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Gly Gly  Leu Ser Glu Ala Asp  Lys Ala Gly Phe Ile  Lys Arg Gln 
    1025                 1030                 1035             


Leu Val  Glu Thr Arg Gln Ile  Thr Lys His Val Ala  Arg Ile Leu 
    1040                 1045                 1050             


Asp Ser  Arg Met Asn Thr Lys  Arg Asp Lys Asn Asp  Lys Pro Ile 
    1055                 1060                 1065             


Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser Lys Leu  Val Ser Asp 
    1070                 1075                 1080             


Phe Arg  Lys Asp Phe Gln Leu  Tyr Lys Val Arg Asp  Ile Asn Asn 
    1085                 1090                 1095             


Tyr His  His Ala His Asp Ala  Tyr Leu Asn Ala Val  Val Gly Thr 
    1100                 1105                 1110             


Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu Ser Glu  Phe Val Tyr 
    1115                 1120                 1125             


Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys Met Ile  Ala Lys Ser 
    1130                 1135                 1140             


Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys Arg Phe  Phe Tyr Ser 
    1145                 1150                 1155             


Asn Ile  Met Asn Phe Phe Lys  Thr Glu Val Lys Leu  Ala Asn Gly 
    1160                 1165                 1170             


Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr Asn Gly  Glu Thr Gly 
    1175                 1180                 1185             


Glu Val  Val Trp Asn Lys Glu  Lys Asp Phe Ala Thr  Val Arg Lys 
    1190                 1195                 1200             


Val Leu  Ala Met Pro Gln Val  Asn Ile Val Lys Lys  Thr Glu Val 
    1205                 1210                 1215             


Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile Leu Ser  Lys Arg Glu 
    1220                 1225                 1230             


Ser Ala  Lys Leu Ile Pro Arg  Lys Lys Gly Trp Asp  Thr Arg Lys 
    1235                 1240                 1245             


Tyr Gly  Gly Phe Gly Ser Pro  Thr Val Ala Tyr Ser  Ile Leu Val 
    1250                 1255                 1260             


Val Ala  Lys Val Glu Lys Gly  Lys Ala Lys Lys Leu  Lys Ser Val 
    1265                 1270                 1275             


Lys Val  Leu Val Gly Ile Thr  Ile Met Glu Lys Gly  Ser Tyr Glu 
    1280                 1285                 1290             


Lys Asp  Pro Ile Gly Phe Leu  Glu Ala Lys Gly Tyr  Lys Asp Ile 
    1295                 1300                 1305             


Lys Lys  Glu Leu Ile Phe Lys  Leu Pro Lys Tyr Ser  Leu Phe Glu 
    1310                 1315                 1320             


Leu Glu  Asn Gly Arg Arg Arg  Met Leu Ala Ser Ala  Thr Glu Leu 
    1325                 1330                 1335             


Gln Lys  Ala Gly Gly Ser Gly  Ser Val Ser Met Val  His Ser Glu 
    1340                 1345                 1350             


Arg Leu  Pro Leu Leu Met Asn  Arg Asn Gly Ile Arg  Phe Gln Ser 
    1355                 1360                 1365             


Glu Val  Ile Asp Pro Asn Pro  Pro Lys Lys Val Lys  Trp Val Leu 
    1370                 1375                 1380             


Thr Gly  Ser Gly Gly Asn Met  Phe Thr Ala Asn Val  Arg Gly Ser 
    1385                 1390                 1395             


Asn Val  Ser Ser Leu Val Lys  His Thr Trp Ser Glu  Ala Phe Val 
    1400                 1405                 1410             


Ser Leu  Ile Glu Asp Arg Gly  Gly Ser Gly Tyr Gln  Thr Val Thr 
    1415                 1420                 1425             


Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu 
    1430                 1435                 1440             


Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg 
    1445                 1450                 1455             


Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1460                 1465                 1470         


<210>  439
<211>  194
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Ancestral FokI mutant

<400>  439

Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
1               5                   10                  15      


Lys Tyr Leu Pro His Glu Tyr Leu Glu Leu Ile Asp Ile Ala Arg Asp 
            20                  25                  30          


Pro Thr Gln Asn Arg Ile Phe Glu Met Lys Thr Met Glu Leu Phe Met 
        35                  40                  45              


Lys Val Tyr Gly Phe Arg Gly Glu His Leu Gly Gly Ser Arg Lys Pro 
    50                  55                  60                  


Asp Gly Ala Ile Tyr Thr Val Gly Leu Pro Ile Asp Tyr Gly Val Ile 
65                  70                  75                  80  


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
                85                  90                  95      


Ala Asp Glu Met Gln Arg Tyr Ile Glu Glu Asn Gln Thr Arg Asn Lys 
            100                 105                 110         


His Ile Asn Pro Asn Glu Trp Trp Lys Val Phe Pro Ser Ser Ile Thr 
        115                 120                 125             


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Phe Lys 
    130                 135                 140                 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Ala 
145                 150                 155                 160 


Leu Asn Val Glu Glu Leu Leu Ile Gly Ala Glu Met Ile Lys Ala Gly 
                165                 170                 175     


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
            180                 185                 190         


Asn Phe 
        


<210>  440
<211>  1625
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, SV40+bipartiteSV40, ancestral mutations in RuvC+REC1/2 
       domain, D4680

<400>  440

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  
    1625 


<210>  441
<211>  1631
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, SV40+bipartiteSV40, ancestral mutations in RuvC+REC1/2 
       domain, 6His tag D4681

<400>  441

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  His His His His His  His 
    1625                 1630     


<210>  442
<211>  1513
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, SV40+bipartiteSV40, ancestral mutations in RuvC+REC1/2 
       domain, HNH deletion D4682

<400>  442

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1505                 1510             


<210>  443
<211>  1519
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, SV40+bipartiteSV40, ancestral mutations in RuvC+REC1/2 
       domain, HNH deletion, 6His tag D4683

<400>  443

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val His His  His His His 
    1505                 1510                 1515             


His 
    


<210>  444
<211>  1625
<212>  PRT
<213>  Artificial Sequence

<220>
<223>   dCasFok, FokI consensus, SV40+bipartite SV40 D4732

<400>  444

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Leu Pro His Glu Tyr Leu Glu Leu Ile Asp Ile Ala 
        35                  40                  45              


Arg Asp Pro Thr Gln Asn Arg Ile Phe Glu Met Lys Thr Met Glu Leu 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Phe Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Leu Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Ile Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Phe Pro Ser Ser 
    130                 135                 140                 


Ile Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Phe Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Ala Leu Asn Val Glu Glu Leu Leu Ile Gly Ala Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  
    1625 


<210>  445
<211>  1631
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI consensus, SV40+bipartiteSV40 D4733

<400>  445

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Leu Pro His Glu Tyr Leu Glu Leu Ile Asp Ile Ala 
        35                  40                  45              


Arg Asp Pro Thr Gln Asn Arg Ile Phe Glu Met Lys Thr Met Glu Leu 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Phe Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Leu Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Ile Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Phe Pro Ser Ser 
    130                 135                 140                 


Ile Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Phe Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Ala Leu Asn Val Glu Glu Leu Leu Ile Gly Ala Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  His His His His His  His 
    1625                 1630     


<210>  446
<211>  1625
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI consensus, SV40+bipartiteSV40, ancestral mutations 
       in RuvC+REC1/2 domain, D4736

<400>  446

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Leu Pro His Glu Tyr Leu Glu Leu Ile Asp Ile Ala 
        35                  40                  45              


Arg Asp Pro Thr Gln Asn Arg Ile Phe Glu Met Lys Thr Met Glu Leu 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Phe Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Leu Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Ile Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Phe Pro Ser Ser 
    130                 135                 140                 


Ile Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Phe Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Ala Leu Asn Val Glu Glu Leu Leu Ile Gly Ala Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  
    1625 


<210>  447
<211>  1631
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI consensus, SV40+bipartiteSV40, ancestral mutations 
       in RuvC+REC1/2 domain, 6His tag D4737

<400>  447

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Leu Pro His Glu Tyr Leu Glu Leu Ile Asp Ile Ala 
        35                  40                  45              


Arg Asp Pro Thr Gln Asn Arg Ile Phe Glu Met Lys Thr Met Glu Leu 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Phe Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Leu Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Ile Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Phe Pro Ser Ser 
    130                 135                 140                 


Ile Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Phe Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Ala Leu Asn Val Glu Glu Leu Leu Ile Gly Ala Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  His His His His His  His 
    1625                 1630     


<210>  448
<211>  1513
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI consensus, SV40+bipartiteSV40, ancestral mutations 
       in RuvC+REC1/2 domain, HNH deletion longer linkers D4740

<400>  448

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Leu Pro His Glu Tyr Leu Glu Leu Ile Asp Ile Ala 
        35                  40                  45              


Arg Asp Pro Thr Gln Asn Arg Ile Phe Glu Met Lys Thr Met Glu Leu 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Phe Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Leu Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Ile Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Phe Pro Ser Ser 
    130                 135                 140                 


Ile Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Phe Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Ala Leu Asn Val Glu Glu Leu Leu Ile Gly Ala Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1505                 1510             


<210>  449
<211>  1519
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI consensus, SV40+bipartiteSV40, ancestral mutations 
       in RuvC+REC1/2 domain, HNH deletion longer linkers, 6His tag 
       D4741

<400>  449

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Leu Pro His Glu Tyr Leu Glu Leu Ile Asp Ile Ala 
        35                  40                  45              


Arg Asp Pro Thr Gln Asn Arg Ile Phe Glu Met Lys Thr Met Glu Leu 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Phe Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Leu Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Ile Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Phe Pro Ser Ser 
    130                 135                 140                 


Ile Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Phe Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Ala Leu Asn Val Glu Glu Leu Leu Ile Gly Ala Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val His His  His His His 
    1505                 1510                 1515             


His 
    


<210>  450
<211>  1615
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       deletion D4694

<400>  450

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Ile Glu Glu Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu 
    1010                 1015                 1020             


Lys Glu  Asn Pro Val Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu 
    1025                 1030                 1035             


Tyr Leu  Tyr Tyr Leu Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln 
    1040                 1045                 1050             


Glu Leu  Asp Ile Asn Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile 
    1055                 1060                 1065             


Val Pro  Gln Ser Phe Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val 
    1070                 1075                 1080             


Leu Thr  Arg Ser Val Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro 
    1085                 1090                 1095             


Ser Glu  Glu Val Val Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu 
    1100                 1105                 1110             


Leu Asn  Ala Lys Leu Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr 
    1115                 1120                 1125             


Lys Ala  Glu Arg Gly Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe 
    1130                 1135                 1140             


Ile Lys  Arg Gln Leu Val Glu  Thr Arg Gln Ile Thr  Lys His Val 
    1145                 1150                 1155             


Ala Arg  Ile Leu Asp Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn 
    1160                 1165                 1170             


Asp Lys  Pro Ile Arg Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys 
    1175                 1180                 1185             


Leu Val  Ser Asp Phe Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg 
    1190                 1195                 1200             


Asp Ile  Asn Asn Tyr His His  Ala His Asp Ala Tyr  Leu Asn Ala 
    1205                 1210                 1215             


Val Val  Gly Thr Ala Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser 
    1220                 1225                 1230             


Glu Phe  Val Tyr Gly Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met 
    1235                 1240                 1245             


Ile Ala  Lys Ser Glu Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg 
    1250                 1255                 1260             


Phe Phe  Tyr Ser Asn Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys 
    1265                 1270                 1275             


Leu Ala  Asn Gly Glu Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn 
    1280                 1285                 1290             


Gly Glu  Thr Gly Glu Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala 
    1295                 1300                 1305             


Thr Val  Arg Lys Val Leu Ala  Met Pro Gln Val Asn  Ile Val Lys 
    1310                 1315                 1320             


Lys Thr  Glu Val Gln Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu 
    1325                 1330                 1335             


Ser Lys  Arg Glu Ser Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp 
    1340                 1345                 1350             


Asp Thr  Arg Lys Tyr Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr 
    1355                 1360                 1365             


Ser Ile  Leu Val Val Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys 
    1370                 1375                 1380             


Leu Lys  Ser Val Lys Val Leu  Val Gly Ile Thr Ile  Met Glu Lys 
    1385                 1390                 1395             


Gly Ser  Tyr Glu Lys Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly 
    1400                 1405                 1410             


Tyr Lys  Asp Ile Lys Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr 
    1415                 1420                 1425             


Ser Leu  Phe Glu Leu Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser 
    1430                 1435                 1440             


Ala Thr  Glu Leu Gln Lys Ala  Asn Glu Leu Val Leu  Pro Gln His 
    1445                 1450                 1455             


Leu Val  Arg Leu Leu Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr 
    1460                 1465                 1470             


Gly Ser  Asn Asn Leu Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe 
    1475                 1480                 1485             


Lys Glu  Ile Phe Glu Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile 
    1490                 1495                 1500             


Leu Lys  Asn Lys Val Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu 
    1505                 1510                 1515             


Gln Phe  Ala Val Ser Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val 
    1520                 1525                 1530             


Ser Leu  Leu Lys Tyr Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr 
    1535                 1540                 1545             


Phe Leu  Asp Leu Asp Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr 
    1550                 1555                 1560             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1565                 1570                 1575             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1580                 1585                 1590             


Ser Gly  Gly Ser Lys Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser 
    1595                 1600                 1605             


Pro Lys  Lys Lys Arg Lys Val  
    1610                 1615 


<210>  451
<211>  1621
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       deletion, 6His tag D4695

<400>  451

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Ile Glu Glu Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu 
    1010                 1015                 1020             


Lys Glu  Asn Pro Val Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu 
    1025                 1030                 1035             


Tyr Leu  Tyr Tyr Leu Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln 
    1040                 1045                 1050             


Glu Leu  Asp Ile Asn Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile 
    1055                 1060                 1065             


Val Pro  Gln Ser Phe Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val 
    1070                 1075                 1080             


Leu Thr  Arg Ser Val Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro 
    1085                 1090                 1095             


Ser Glu  Glu Val Val Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu 
    1100                 1105                 1110             


Leu Asn  Ala Lys Leu Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr 
    1115                 1120                 1125             


Lys Ala  Glu Arg Gly Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe 
    1130                 1135                 1140             


Ile Lys  Arg Gln Leu Val Glu  Thr Arg Gln Ile Thr  Lys His Val 
    1145                 1150                 1155             


Ala Arg  Ile Leu Asp Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn 
    1160                 1165                 1170             


Asp Lys  Pro Ile Arg Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys 
    1175                 1180                 1185             


Leu Val  Ser Asp Phe Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg 
    1190                 1195                 1200             


Asp Ile  Asn Asn Tyr His His  Ala His Asp Ala Tyr  Leu Asn Ala 
    1205                 1210                 1215             


Val Val  Gly Thr Ala Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser 
    1220                 1225                 1230             


Glu Phe  Val Tyr Gly Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met 
    1235                 1240                 1245             


Ile Ala  Lys Ser Glu Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg 
    1250                 1255                 1260             


Phe Phe  Tyr Ser Asn Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys 
    1265                 1270                 1275             


Leu Ala  Asn Gly Glu Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn 
    1280                 1285                 1290             


Gly Glu  Thr Gly Glu Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala 
    1295                 1300                 1305             


Thr Val  Arg Lys Val Leu Ala  Met Pro Gln Val Asn  Ile Val Lys 
    1310                 1315                 1320             


Lys Thr  Glu Val Gln Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu 
    1325                 1330                 1335             


Ser Lys  Arg Glu Ser Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp 
    1340                 1345                 1350             


Asp Thr  Arg Lys Tyr Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr 
    1355                 1360                 1365             


Ser Ile  Leu Val Val Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys 
    1370                 1375                 1380             


Leu Lys  Ser Val Lys Val Leu  Val Gly Ile Thr Ile  Met Glu Lys 
    1385                 1390                 1395             


Gly Ser  Tyr Glu Lys Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly 
    1400                 1405                 1410             


Tyr Lys  Asp Ile Lys Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr 
    1415                 1420                 1425             


Ser Leu  Phe Glu Leu Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser 
    1430                 1435                 1440             


Ala Thr  Glu Leu Gln Lys Ala  Asn Glu Leu Val Leu  Pro Gln His 
    1445                 1450                 1455             


Leu Val  Arg Leu Leu Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr 
    1460                 1465                 1470             


Gly Ser  Asn Asn Leu Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe 
    1475                 1480                 1485             


Lys Glu  Ile Phe Glu Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile 
    1490                 1495                 1500             


Leu Lys  Asn Lys Val Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu 
    1505                 1510                 1515             


Gln Phe  Ala Val Ser Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val 
    1520                 1525                 1530             


Ser Leu  Leu Lys Tyr Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr 
    1535                 1540                 1545             


Phe Leu  Asp Leu Asp Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr 
    1550                 1555                 1560             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1565                 1570                 1575             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1580                 1585                 1590             


Ser Gly  Gly Ser Lys Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser 
    1595                 1600                 1605             


Pro Lys  Lys Lys Arg Lys Val  His His His His His  His 
    1610                 1615                 1620     


<210>  452
<211>  1503
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       deletion, HNHdeletion D4696

<400>  452

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1430                 1435                 1440             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1445                 1450                 1455             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1460                 1465                 1470             


Asp Leu  Ser Gln Leu Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr 
    1475                 1480                 1485             


Ala Asp  Gly Ser Glu Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1490                 1495                 1500             


<210>  453
<211>  1509
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       deletion, HNHdeletion, 6His tag D4697

<400>  453

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1430                 1435                 1440             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1445                 1450                 1455             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1460                 1465                 1470             


Asp Leu  Ser Gln Leu Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr 
    1475                 1480                 1485             


Ala Asp  Gly Ser Glu Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1490                 1495                 1500             


His His  His His His His 
    1505                 


<210>  454
<211>  1509
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       deletion, HNHdeletion, 6His tag D4698

<400>  454

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ala Thr Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Lys Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Leu Thr Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1430                 1435                 1440             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1445                 1450                 1455             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1460                 1465                 1470             


Asp Leu  Ser Gln Leu Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr 
    1475                 1480                 1485             


Ala Asp  Gly Ser Glu Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1490                 1495                 1500             


His His  His His His His 
    1505                 


<210>  455
<211>  1519
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       QQmutant, HNHdeletion, 6His tag D4699

<400>  455

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val His His  His His His 
    1505                 1510                 1515             


His 
    


<210>  456
<211>  1519
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  ScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       AAmutant, HNHdeletion, 6His tag D4700

<400>  456

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val His His  His His His 
    1505                 1510                 1515             


His 
    


<210>  457
<211>  1509
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop Sp 
       Replacement, HNHdeletion, 6His tag D4701

<400>  457

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1430                 1435                 1440             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1445                 1450                 1455             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1460                 1465                 1470             


Asp Leu  Ser Gln Leu Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr 
    1475                 1480                 1485             


Ala Asp  Gly Ser Glu Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1490                 1495                 1500             


His His  His His His His 
    1505                 


<210>  458
<211>  1509
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  > dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       deletion, HNHdeletion, 6His tag D4702

<400>  458

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Ser Gly Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys 
    1430                 1435                 1440             


Gln Gly  Arg Leu Arg Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1445                 1450                 1455             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1460                 1465                 1470             


Asp Leu  Ser Gln Leu Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr 
    1475                 1480                 1485             


Ala Asp  Gly Ser Glu Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1490                 1495                 1500             


His His  His His His His 
    1505                 


<210>  459
<211>  1519
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       QQmutant, HNHdeletion, 6His tag D4703

<400>  459

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val His His  His His His 
    1505                 1510                 1515             


His 
    


<210>  460
<211>  1519
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+bipartiteSV40, ancestral mutations, Scloop 
       AAmutant, HNHdeletion, 6His tag D4704

<400>  460

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val His His  His His His 
    1505                 1510                 1515             


His 
    


<210>  461
<211>  1526
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       deletion, HNH deletion, PAM replaced with Zinc finger D4710

<400>  461

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Gly Gly Ser Ala  Gly Gly Thr Gly Gly  Ser Ala Gly 
    1430                 1435                 1440             


Gly Ser  Gly Ala Pro Tyr Arg  Cys Pro Cys Arg Ala  Gly Cys Pro 
    1445                 1450                 1455             


Ser Leu  Ala Ser Met Gln Ala  His Met Arg Gly His  Ser Gly Gly 
    1460                 1465                 1470             


Ser Gly  Tyr Gln Thr Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile 
    1475                 1480                 1485             


Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser 
    1490                 1495                 1500             


Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly 
    1505                 1510                 1515             


Gln Ala  Lys Lys Lys Lys Gly  Ala 
    1520                 1525     


<210>  462
<211>  1443
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       deletion, HNH deletion, PAM replaced with LacI DNA binding domain
       D4711

<400>  462

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Gly Gly  Ser Gly Met Lys Pro  Val Thr Leu Tyr Asp  Val Ala Glu 
    1340                 1345                 1350             


Tyr Ala  Gly Val Ser Tyr Gln  Thr Val Ser Arg Val  Val Asn Gln 
    1355                 1360                 1365             


Ala Ser  His Val Ser Ala Lys  Thr Arg Glu Lys Val  Glu Ala Ala 
    1370                 1375                 1380             


Met Ala  Glu Leu Asn Gly Gly  Ser Gly Tyr Gln Thr  Val Thr Glu 
    1385                 1390                 1395             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1400                 1405                 1410             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1415                 1420                 1425             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1430                 1435                 1440             


<210>  463
<211>  1463
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       deletion, HNH deletion, PAM replaced with SSO7D D4712

<400>  463

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Gly Gly  Ser Ala Gly Met Ala  Thr Val Lys Phe Lys  Tyr Lys Gly 
    1340                 1345                 1350             


Glu Glu  Lys Glu Val Asp Ile  Ser Lys Ile Lys Lys  Val Trp Arg 
    1355                 1360                 1365             


Val Gly  Lys Met Ile Ser Phe  Thr Tyr Asp Glu Gly  Gly Gly Lys 
    1370                 1375                 1380             


Thr Gly  Arg Gly Ala Val Ser  Glu Lys Asp Ala Pro  Lys Glu Leu 
    1385                 1390                 1395             


Leu Gln  Met Leu Glu Lys Gln  Lys Lys Gly Gly Ser  Ala Gly Tyr 
    1400                 1405                 1410             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1415                 1420                 1425             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1430                 1435                 1440             


Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys 
    1445                 1450                 1455             


Lys Lys  Lys Gly Ala 
    1460             


<210>  464
<211>  1587
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       deletion, HNH deletion, PAM replaced with HMGN D4713

<400>  464

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Gly Gly Ser Gly  Pro Lys Arg Lys Val  Ser Ser Ala 
    1430                 1435                 1440             


Glu Gly  Ala Ala Lys Glu Glu  Pro Lys Arg Arg Ser  Ala Arg Leu 
    1445                 1450                 1455             


Ser Ala  Lys Pro Pro Ala Lys  Val Glu Ala Lys Pro  Lys Lys Ala 
    1460                 1465                 1470             


Ala Ala  Lys Asp Lys Ser Ser  Asp Lys Lys Val Gln  Thr Lys Gly 
    1475                 1480                 1485             


Lys Arg  Gly Ala Lys Gly Lys  Gln Ala Glu Val Ala  Asn Gln Glu 
    1490                 1495                 1500             


Thr Lys  Glu Asp Leu Pro Ala  Glu Asn Gly Glu Thr  Lys Thr Glu 
    1505                 1510                 1515             


Glu Ser  Pro Ala Ser Asp Glu  Ala Gly Glu Lys Glu  Ala Lys Ser 
    1520                 1525                 1530             


Asp Gly  Gly Ser Gly Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1535                 1540                 1545             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1550                 1555                 1560             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1565                 1570                 1575             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1580                 1585         


<210>  465
<211>  1483
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       deletion, HNH deletion, PAM replaced with STO7 D4714

<400>  465

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Thr Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Ala Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Glu 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser Ala Gly  Gly Val Thr 
    1340                 1345                 1350             


Val Lys  Phe Lys Tyr Lys Gly  Glu Glu Lys Glu Val  Asp Ile Ser 
    1355                 1360                 1365             


Lys Ile  Lys Lys Val Trp Arg  Val Gly Lys Met Ile  Ser Phe Thr 
    1370                 1375                 1380             


Tyr Asp  Asp Asn Gly Lys Thr  Gly Arg Gly Ala Val  Ser Glu Lys 
    1385                 1390                 1395             


Asp Ala  Pro Lys Glu Leu Leu  Gln Met Leu Glu Lys  Ser Gly Lys 
    1400                 1405                 1410             


Lys Gly  Gly Ser Ala Gly Gly  Thr Gly Gly Ser Ala  Gly Gly Tyr 
    1415                 1420                 1425             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1430                 1435                 1440             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1445                 1450                 1455             


Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys 
    1460                 1465                 1470             


Lys Lys  Lys Pro Lys Lys Lys  Arg Lys Val 
    1475                 1480             


<210>  466
<211>  1536
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       QQmutation, HNH deletion, PAM replaced with Zinc finger D4715

<400>  466

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly 
    1430                 1435                 1440             


Ser Ala  Gly Gly Thr Gly Gly  Ser Ala Gly Gly Ser  Gly Ala Pro 
    1445                 1450                 1455             


Tyr Arg  Cys Pro Cys Arg Ala  Gly Cys Pro Ser Leu  Ala Ser Met 
    1460                 1465                 1470             


Gln Ala  His Met Arg Gly His  Ser Gly Gly Ser Gly  Tyr Gln Thr 
    1475                 1480                 1485             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1490                 1495                 1500             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1505                 1510                 1515             


Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys 
    1520                 1525                 1530             


Lys Gly  Ala 
    1535     


<210>  467
<211>  1453
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       QQmutation HNH deletion, PAM replaced with LacI DNA binding 
       domain D4716

<400>  467

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Gly Met 
    1340                 1345                 1350             


Lys Pro  Val Thr Leu Tyr Asp  Val Ala Glu Tyr Ala  Gly Val Ser 
    1355                 1360                 1365             


Tyr Gln  Thr Val Ser Arg Val  Val Asn Gln Ala Ser  His Val Ser 
    1370                 1375                 1380             


Ala Lys  Thr Arg Glu Lys Val  Glu Ala Ala Met Ala  Glu Leu Asn 
    1385                 1390                 1395             


Gly Gly  Ser Gly Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1400                 1405                 1410             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1415                 1420                 1425             


Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys 
    1430                 1435                 1440             


Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1445                 1450             


<210>  468
<211>  1473
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       QQmutation, HNH deletion, PAM replaced with SSO7D D4717

<400>  468

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly 
    1340                 1345                 1350             


Met Ala  Thr Val Lys Phe Lys  Tyr Lys Gly Glu Glu  Lys Glu Val 
    1355                 1360                 1365             


Asp Ile  Ser Lys Ile Lys Lys  Val Trp Arg Val Gly  Lys Met Ile 
    1370                 1375                 1380             


Ser Phe  Thr Tyr Asp Glu Gly  Gly Gly Lys Thr Gly  Arg Gly Ala 
    1385                 1390                 1395             


Val Ser  Glu Lys Asp Ala Pro  Lys Glu Leu Leu Gln  Met Leu Glu 
    1400                 1405                 1410             


Lys Gln  Lys Lys Gly Gly Ser  Ala Gly Tyr Gln Thr  Val Thr Glu 
    1415                 1420                 1425             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1430                 1435                 1440             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1445                 1450                 1455             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1460                 1465                 1470             


<210>  469
<211>  1597
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       QQmutation, HNH deletion, PAM replaced with HMGN D4718

<400>  469

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly 
    1430                 1435                 1440             


Ser Gly  Pro Lys Arg Lys Val  Ser Ser Ala Glu Gly  Ala Ala Lys 
    1445                 1450                 1455             


Glu Glu  Pro Lys Arg Arg Ser  Ala Arg Leu Ser Ala  Lys Pro Pro 
    1460                 1465                 1470             


Ala Lys  Val Glu Ala Lys Pro  Lys Lys Ala Ala Ala  Lys Asp Lys 
    1475                 1480                 1485             


Ser Ser  Asp Lys Lys Val Gln  Thr Lys Gly Lys Arg  Gly Ala Lys 
    1490                 1495                 1500             


Gly Lys  Gln Ala Glu Val Ala  Asn Gln Glu Thr Lys  Glu Asp Leu 
    1505                 1510                 1515             


Pro Ala  Glu Asn Gly Glu Thr  Lys Thr Glu Glu Ser  Pro Ala Ser 
    1520                 1525                 1530             


Asp Glu  Ala Gly Glu Lys Glu  Ala Lys Ser Asp Gly  Gly Ser Gly 
    1535                 1540                 1545             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1550                 1555                 1560             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1565                 1570                 1575             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1580                 1585                 1590             


Lys Lys  Lys Lys 
    1595         


<210>  470
<211>  1493
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       QQmutation, HNH deletion, PAM replaced with STO7 D4719

<400>  470

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Gln Lys Gln Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly 
    1340                 1345                 1350             


Gly Thr  Gly Gly Ser Ala Gly  Gly Val Thr Val Lys  Phe Lys Tyr 
    1355                 1360                 1365             


Lys Gly  Glu Glu Lys Glu Val  Asp Ile Ser Lys Ile  Lys Lys Val 
    1370                 1375                 1380             


Trp Arg  Val Gly Lys Met Ile  Ser Phe Thr Tyr Asp  Asp Asn Gly 
    1385                 1390                 1395             


Lys Thr  Gly Arg Gly Ala Val  Ser Glu Lys Asp Ala  Pro Lys Glu 
    1400                 1405                 1410             


Leu Leu  Gln Met Leu Glu Lys  Ser Gly Lys Lys Gly  Gly Ser Ala 
    1415                 1420                 1425             


Gly Gly  Thr Gly Gly Ser Ala  Gly Gly Tyr Gln Thr  Val Thr Glu 
    1430                 1435                 1440             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1445                 1450                 1455             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1460                 1465                 1470             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Pro Lys 
    1475                 1480                 1485             


Lys Lys  Arg Lys Val 
    1490             


<210>  471
<211>  1536
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       AAmutation, HNH deletion, PAM replaced with Zinc finger D4720

<400>  471

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly 
    1430                 1435                 1440             


Ser Ala  Gly Gly Thr Gly Gly  Ser Ala Gly Gly Ser  Gly Ala Pro 
    1445                 1450                 1455             


Tyr Arg  Cys Pro Cys Arg Ala  Gly Cys Pro Ser Leu  Ala Ser Met 
    1460                 1465                 1470             


Gln Ala  His Met Arg Gly His  Ser Gly Gly Ser Gly  Tyr Gln Thr 
    1475                 1480                 1485             


Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr 
    1490                 1495                 1500             


Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1505                 1510                 1515             


Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly Gln Ala  Lys Lys Lys 
    1520                 1525                 1530             


Lys Gly  Ala 
    1535     


<210>  472
<211>  1453
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       AAmutation HNH deletion, PAM replaced with LacI DNA binding 
       domain D4721

<400>  472

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Gly Met 
    1340                 1345                 1350             


Lys Pro  Val Thr Leu Tyr Asp  Val Ala Glu Tyr Ala  Gly Val Ser 
    1355                 1360                 1365             


Tyr Gln  Thr Val Ser Arg Val  Val Asn Gln Ala Ser  His Val Ser 
    1370                 1375                 1380             


Ala Lys  Thr Arg Glu Lys Val  Glu Ala Ala Met Ala  Glu Leu Asn 
    1385                 1390                 1395             


Gly Gly  Ser Gly Tyr Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr 
    1400                 1405                 1410             


Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp 
    1415                 1420                 1425             


Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys 
    1430                 1435                 1440             


Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1445                 1450             


<210>  473
<211>  1473
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       AAmutation, HNH deletion, PAM replaced with SSO7D D4722

<400>  473

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly 
    1340                 1345                 1350             


Met Ala  Thr Val Lys Phe Lys  Tyr Lys Gly Glu Glu  Lys Glu Val 
    1355                 1360                 1365             


Asp Ile  Ser Lys Ile Lys Lys  Val Trp Arg Val Gly  Lys Met Ile 
    1370                 1375                 1380             


Ser Phe  Thr Tyr Asp Glu Gly  Gly Gly Lys Thr Gly  Arg Gly Ala 
    1385                 1390                 1395             


Val Ser  Glu Lys Asp Ala Pro  Lys Glu Leu Leu Gln  Met Leu Glu 
    1400                 1405                 1410             


Lys Gln  Lys Lys Gly Gly Ser  Ala Gly Tyr Gln Thr  Val Thr Glu 
    1415                 1420                 1425             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1430                 1435                 1440             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1445                 1450                 1455             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1460                 1465                 1470             


<210>  474
<211>  1597
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       AAmutation, HNH deletion, PAM replaced with HMGN D4723

<400>  474

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Gly Gly 
    1430                 1435                 1440             


Ser Gly  Pro Lys Arg Lys Val  Ser Ser Ala Glu Gly  Ala Ala Lys 
    1445                 1450                 1455             


Glu Glu  Pro Lys Arg Arg Ser  Ala Arg Leu Ser Ala  Lys Pro Pro 
    1460                 1465                 1470             


Ala Lys  Val Glu Ala Lys Pro  Lys Lys Ala Ala Ala  Lys Asp Lys 
    1475                 1480                 1485             


Ser Ser  Asp Lys Lys Val Gln  Thr Lys Gly Lys Arg  Gly Ala Lys 
    1490                 1495                 1500             


Gly Lys  Gln Ala Glu Val Ala  Asn Gln Glu Thr Lys  Glu Asp Leu 
    1505                 1510                 1515             


Pro Ala  Glu Asn Gly Glu Thr  Lys Thr Glu Glu Ser  Pro Ala Ser 
    1520                 1525                 1530             


Asp Glu  Ala Gly Glu Lys Glu  Ala Lys Ser Asp Gly  Gly Ser Gly 
    1535                 1540                 1545             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1550                 1555                 1560             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1565                 1570                 1575             


Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys Lys Ala  Gly Gln Ala 
    1580                 1585                 1590             


Lys Lys  Lys Lys 
    1595         


<210>  475
<211>  1493
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       AAmutation, HNH deletion, PAM GAreplaced with STO7 D4724

<400>  475

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Ala Lys Ala Thr Thr Lys Leu Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Gly Gly  Ser Ala Gly 
    1340                 1345                 1350             


Gly Thr  Gly Gly Ser Ala Gly  Gly Val Thr Val Lys  Phe Lys Tyr 
    1355                 1360                 1365             


Lys Gly  Glu Glu Lys Glu Val  Asp Ile Ser Lys Ile  Lys Lys Val 
    1370                 1375                 1380             


Trp Arg  Val Gly Lys Met Ile  Ser Phe Thr Tyr Asp  Asp Asn Gly 
    1385                 1390                 1395             


Lys Thr  Gly Arg Gly Ala Val  Ser Glu Lys Asp Ala  Pro Lys Glu 
    1400                 1405                 1410             


Leu Leu  Gln Met Leu Glu Lys  Ser Gly Lys Lys Gly  Gly Ser Ala 
    1415                 1420                 1425             


Gly Gly  Thr Gly Gly Ser Ala  Gly Gly Tyr Gln Thr  Val Thr Glu 
    1430                 1435                 1440             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1445                 1450                 1455             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1460                 1465                 1470             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Pro Lys 
    1475                 1480                 1485             


Lys Lys  Arg Lys Val 
    1490             


<210>  476
<211>  1526
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       SpReplacement, HNH deletion, PAM replaced with Zinc finger D4725

<400>  476

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Gly Gly Ser Ala  Gly Gly Thr Gly Gly  Ser Ala Gly 
    1430                 1435                 1440             


Gly Ser  Gly Ala Pro Tyr Arg  Cys Pro Cys Arg Ala  Gly Cys Pro 
    1445                 1450                 1455             


Ser Leu  Ala Ser Met Gln Ala  His Met Arg Gly His  Ser Gly Gly 
    1460                 1465                 1470             


Ser Gly  Tyr Gln Thr Val Thr  Glu Val Leu Asp Ala  Thr Leu Ile 
    1475                 1480                 1485             


Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr Arg Thr  Asp Leu Ser 
    1490                 1495                 1500             


Gln Leu  Gly Gly Asp Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly 
    1505                 1510                 1515             


Gln Ala  Lys Lys Lys Lys Gly  Ala 
    1520                 1525     


<210>  477
<211>  1443
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       SpReplacement HNH deletion, PAM replaced with LacI DNA binding 
       domain D4726

<400>  477

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Gly Gly  Ser Gly Met Lys Pro  Val Thr Leu Tyr Asp  Val Ala Glu 
    1340                 1345                 1350             


Tyr Ala  Gly Val Ser Tyr Gln  Thr Val Ser Arg Val  Val Asn Gln 
    1355                 1360                 1365             


Ala Ser  His Val Ser Ala Lys  Thr Arg Glu Lys Val  Glu Ala Ala 
    1370                 1375                 1380             


Met Ala  Glu Leu Asn Gly Gly  Ser Gly Tyr Gln Thr  Val Thr Glu 
    1385                 1390                 1395             


Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser Ile Thr  Gly Leu Tyr 
    1400                 1405                 1410             


Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly Gly Asp  Lys Arg Pro 
    1415                 1420                 1425             


Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys Lys Lys  Lys Gly Ala 
    1430                 1435                 1440             


<210>  478
<211>  1463
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       SpReplacement HNH deletion, PAM replaced with SSO7D D4727

<400>  478

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Gly Gly  Ser Ala Gly Met Ala  Thr Val Lys Phe Lys  Tyr Lys Gly 
    1340                 1345                 1350             


Glu Glu  Lys Glu Val Asp Ile  Ser Lys Ile Lys Lys  Val Trp Arg 
    1355                 1360                 1365             


Val Gly  Lys Met Ile Ser Phe  Thr Tyr Asp Glu Gly  Gly Gly Lys 
    1370                 1375                 1380             


Thr Gly  Arg Gly Ala Val Ser  Glu Lys Asp Ala Pro  Lys Glu Leu 
    1385                 1390                 1395             


Leu Gln  Met Leu Glu Lys Gln  Lys Lys Gly Gly Ser  Ala Gly Tyr 
    1400                 1405                 1410             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1415                 1420                 1425             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1430                 1435                 1440             


Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys 
    1445                 1450                 1455             


Lys Lys  Lys Gly Ala 
    1460             


<210>  479
<211>  1587
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       SpReplacement HNH deletion, PAM replaced with HMGN D4728

<400>  479

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Asn Glu  Leu Val Leu Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr 
    1340                 1345                 1350             


Thr Gln  Asn Ile Ser Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr 
    1355                 1360                 1365             


Ile Glu  Gln His Arg Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile 
    1370                 1375                 1380             


Ile Asp  Phe Ser Glu Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser 
    1385                 1390                 1395             


Asn Leu  Lys Ser Ser Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser 
    1400                 1405                 1410             


Ile Leu  Leu Ser Asn Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser 
    1415                 1420                 1425             


Phe Gly  Ala Gly Gly Ser Gly  Pro Lys Arg Lys Val  Ser Ser Ala 
    1430                 1435                 1440             


Glu Gly  Ala Ala Lys Glu Glu  Pro Lys Arg Arg Ser  Ala Arg Leu 
    1445                 1450                 1455             


Ser Ala  Lys Pro Pro Ala Lys  Val Glu Ala Lys Pro  Lys Lys Ala 
    1460                 1465                 1470             


Ala Ala  Lys Asp Lys Ser Ser  Asp Lys Lys Val Gln  Thr Lys Gly 
    1475                 1480                 1485             


Lys Arg  Gly Ala Lys Gly Lys  Gln Ala Glu Val Ala  Asn Gln Glu 
    1490                 1495                 1500             


Thr Lys  Glu Asp Leu Pro Ala  Glu Asn Gly Glu Thr  Lys Thr Glu 
    1505                 1510                 1515             


Glu Ser  Pro Ala Ser Asp Glu  Ala Gly Glu Lys Glu  Ala Lys Ser 
    1520                 1525                 1530             


Asp Gly  Gly Ser Gly Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala 
    1535                 1540                 1545             


Thr Leu  Ile Tyr Gln Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr 
    1550                 1555                 1560             


Asp Leu  Ser Gln Leu Gly Gly  Asp Lys Arg Pro Ala  Ala Thr Lys 
    1565                 1570                 1575             


Lys Ala  Gly Gln Ala Lys Lys  Lys Lys 
    1580                 1585         


<210>  480
<211>  1483
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dScCasFok, SV40+nucleoplasmin, ancestral mutations, Scloop 
       SpReplacement HNH deletion, PAM replaced with STO7 D4729

<400>  480

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
        595                 600                 605             


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
    610                 615                 620                 


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
625                 630                 635                 640 


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
                645                 650                 655     


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
            660                 665                 670         


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
        675                 680                 685             


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
    690                 695                 700                 


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
705                 710                 715                 720 


Glu Arg Met Thr Asn Phe Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu 
                725                 730                 735     


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
            740                 745                 750         


Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Glu Phe Leu 
        755                 760                 765             


Ser Gly Glu Gln Lys Glu Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
    770                 775                 780                 


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
785                 790                 795                 800 


Glu Cys Phe Asp Ser Val Glu Ile Ile Gly Val Glu Asp Arg Phe Asn 
                805                 810                 815     


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
            820                 825                 830         


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
        835                 840                 845             


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
    850                 855                 860                 


Lys Lys Tyr Ala Asn Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
865                 870                 875                 880 


Arg Arg His Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Met Ile Asn 
                885                 890                 895     


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
            900                 905                 910         


Ser Asp Gly Phe Ser Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
        915                 920                 925             


Ser Leu Thr Phe Lys Glu Glu Ile Glu Lys Ala Gln Val Ser Gly Gln 
    930                 935                 940                 


Gly Asp Ser Leu His Glu Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala 
945                 950                 955                 960 


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Ile Val Asp Glu Leu Val 
                965                 970                 975     


Lys Val Met Gly His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg 
            980                 985                 990         


Glu Asn Gln Thr Thr Thr Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys 
        995                 1000                 1005             


Lys Arg  Gly Gly Ser Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu 
    1010                 1015                 1020             


Ser Glu  Ala Asp Lys Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu 
    1025                 1030                 1035             


Thr Arg  Gln Ile Thr Lys His  Val Ala Arg Ile Leu  Asp Ser Arg 
    1040                 1045                 1050             


Met Asn  Thr Lys Arg Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val 
    1055                 1060                 1065             


Lys Val  Ile Thr Leu Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys 
    1070                 1075                 1080             


Asp Phe  Gln Leu Tyr Lys Val  Arg Asp Ile Asn Asn  Tyr His His 
    1085                 1090                 1095             


Ala His  Asp Ala Tyr Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile 
    1100                 1105                 1110             


Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr 
    1115                 1120                 1125             


Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu 
    1130                 1135                 1140             


Ile Gly  Lys Ala Thr Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met 
    1145                 1150                 1155             


Asn Phe  Phe Lys Thr Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg 
    1160                 1165                 1170             


Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val 
    1175                 1180                 1185             


Trp Asn  Lys Glu Lys Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala 
    1190                 1195                 1200             


Met Pro  Gln Val Asn Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly 
    1205                 1210                 1215             


Gly Phe  Ser Lys Glu Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys 
    1220                 1225                 1230             


Leu Ile  Pro Arg Lys Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly 
    1235                 1240                 1245             


Phe Gly  Ser Pro Thr Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys 
    1250                 1255                 1260             


Val Glu  Lys Gly Lys Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu 
    1265                 1270                 1275             


Val Gly  Ile Thr Ile Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro 
    1280                 1285                 1290             


Ile Gly  Phe Leu Glu Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu 
    1295                 1300                 1305             


Leu Ile  Phe Lys Leu Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn 
    1310                 1315                 1320             


Gly Arg  Arg Arg Met Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala 
    1325                 1330                 1335             


Gly Gly  Ser Ala Gly Gly Thr  Gly Gly Ser Ala Gly  Gly Val Thr 
    1340                 1345                 1350             


Val Lys  Phe Lys Tyr Lys Gly  Glu Glu Lys Glu Val  Asp Ile Ser 
    1355                 1360                 1365             


Lys Ile  Lys Lys Val Trp Arg  Val Gly Lys Met Ile  Ser Phe Thr 
    1370                 1375                 1380             


Tyr Asp  Asp Asn Gly Lys Thr  Gly Arg Gly Ala Val  Ser Glu Lys 
    1385                 1390                 1395             


Asp Ala  Pro Lys Glu Leu Leu  Gln Met Leu Glu Lys  Ser Gly Lys 
    1400                 1405                 1410             


Lys Gly  Gly Ser Ala Gly Gly  Thr Gly Gly Ser Ala  Gly Gly Tyr 
    1415                 1420                 1425             


Gln Thr  Val Thr Glu Val Leu  Asp Ala Thr Leu Ile  Tyr Gln Ser 
    1430                 1435                 1440             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Thr Asp Leu Ser  Gln Leu Gly 
    1445                 1450                 1455             


Gly Asp  Lys Arg Pro Ala Ala  Thr Lys Lys Ala Gly  Gln Ala Lys 
    1460                 1465                 1470             


Lys Lys  Lys Pro Lys Lys Lys  Arg Lys Val 
    1475                 1480             


<210>  481
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  short positively charged linker

<400>  481

Gly Lys Ser Lys Gly 
1               5   


<210>  482
<211>  100
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Residues 1-100 of HMGN

<400>  482

Met Pro Lys Arg Lys Val Ser Ser Ala Glu Gly Ala Ala Lys Glu Glu 
1               5                   10                  15      


Pro Lys Arg Arg Ser Ala Arg Leu Ser Ala Lys Pro Pro Ala Lys Val 
            20                  25                  30          


Glu Ala Lys Pro Lys Lys Ala Ala Ala Lys Asp Lys Ser Ser Asp Lys 
        35                  40                  45              


Lys Val Gln Thr Lys Gly Lys Arg Gly Ala Lys Gly Lys Gln Ala Glu 
    50                  55                  60                  


Val Ala Asn Gln Glu Thr Lys Glu Asp Leu Pro Ala Glu Asn Gly Glu 
65                  70                  75                  80  


Thr Lys Thr Glu Glu Ser Pro Ala Ser Asp Glu Ala Gly Glu Lys Glu 
                85                  90                  95      


Ala Lys Ser Asp 
            100 


<210>  483
<211>  83
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Residues 1-84 of HMGB1

<400>  483

Gly Lys Gly Asp Pro Lys Lys Pro Arg Gly Lys Met Ser Ser Tyr Ala 
1               5                   10                  15      


Phe Phe Val Gln Thr Cys Arg Glu Glu His Lys Lys Lys His Pro Asp 
            20                  25                  30          


Ala Ser Val Asn Phe Ser Glu Phe Ser Lys Lys Cys Ser Glu Arg Trp 
        35                  40                  45              


Lys Thr Met Ser Ala Lys Glu Lys Gly Lys Phe Glu Asp Met Ala Lys 
    50                  55                  60                  


Ala Asp Lys Ala Arg Tyr Glu Arg Glu Met Lys Thr Tyr Ile Pro Pro 
65                  70                  75                  80  


Lys Gly Glu 
            


<210>  484
<211>  79
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Residues 1-79 of HMGB3

<400>  484

Met Ala Lys Gly Asp Pro Lys Lys Pro Lys Gly Lys Met Ser Ala Tyr 
1               5                   10                  15      


Ala Phe Phe Val Gln Thr Cys Arg Glu Glu His Lys Lys Lys Asn Pro 
            20                  25                  30          


Glu Val Pro Val Asn Phe Ala Glu Phe Ser Lys Lys Cys Ser Glu Arg 
        35                  40                  45              


Trp Lys Thr Met Ser Gly Lys Glu Lys Ser Lys Phe Asp Glu Met Ala 
    50                  55                  60                  


Lys Ala Asp Lys Val Arg Tyr Asp Arg Glu Met Lys Asp Tyr Gly 
65                  70                  75                  


<210>  485
<211>  15
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HNS linker

<400>  485

Ala Ala Val Lys Ser Gly Thr Lys Ala Lys Arg Ala Gln Arg Pro 
1               5                   10                  15  


<210>  486
<211>  194
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Enhanced FokI nuclease domain

<400>  486

Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu 
1               5                   10                  15      


Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn 
            20                  25                  30          


Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met 
        35                  40                  45              


Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg Lys Pro 
    50                  55                  60                  


Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile 
65                  70                  75                  80  


Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln 
                85                  90                  95      


Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys 
            100                 105                 110         


His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr 
        115                 120                 125             


Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys 
    130                 135                 140                 


Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val 
145                 150                 155                 160 


Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly 
                165                 170                 175     


Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile 
            180                 185                 190         


Asn Phe 
        


<210>  487
<211>  1625
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI enhanced, SV40+bipartiteSV40

<400>  487

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  
    1625 


<210>  488
<211>  1631
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI enhanced, SV40+bipartiteSV40, 6His tag

<400>  488

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Glu Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asn Arg Lys Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Met Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Arg Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe Gln Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys Asn Glu Arg His Pro Ile Phe Gly Asn Leu Ala Asp 
            340                 345                 350         


Glu Val Ala Tyr His Arg Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Pro Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Ile Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Lys Leu Asn Ala Glu Asn Ser Asp Val Ala Lys Leu Phe Tyr Gln Leu 
                405                 410                 415     


Ile Gln Thr Tyr Asn Gln Leu Phe Glu Glu Ser Pro Leu Asp Glu Ile 
            420                 425                 430         


Glu Val Asp Ala Lys Gly Ile Leu Ser Ala Arg Leu Ser Lys Ser Lys 
        435                 440                 445             


Arg Leu Glu Lys Leu Ile Ala Val Phe Pro Asn Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Ser Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Ser Asn Ser Glu Val Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Phe Pro Glu Lys Tyr Ala 
                565                 570                 575     


Glu Ile Phe Lys Asp Asp Thr Lys Asn Gly Tyr Ala Gly Tyr Val Gly 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Leu Ala Thr Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Asp Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Lys Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Glu Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Leu Thr 
    690                 695                 700                 


Arg Lys Ser Glu Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Gln Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Arg Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Lys 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  His His His His His  His 
    1625                 1630     


<210>  489
<211>  1625
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI enhanced, SV40+bipartiteSV40, ancestral mutations 
       in RuvC+REC1/2 domain

<400>  489

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  
    1625 


<210>  490
<211>  1631
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI enhanced, SV40+bipartiteSV40, ancestral mutations 
       in RuvC+REC1/2 domain, 6His tag

<400>  490

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Ile Glu Glu 
    1010                 1015                 1020             


Gly Ile  Lys Glu Leu Glu Ser  Gln Ile Leu Lys Glu  Asn Pro Val 
    1025                 1030                 1035             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1040                 1045                 1050             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1055                 1060                 1065             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1070                 1075                 1080             


Ile Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Val 
    1085                 1090                 1095             


Glu Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1100                 1105                 1110             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1115                 1120                 1125             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1130                 1135                 1140             


Gly Leu  Ser Glu Ala Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1145                 1150                 1155             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Arg  Ile Leu Asp 
    1160                 1165                 1170             


Ser Arg  Met Asn Thr Lys Arg  Asp Lys Asn Asp Lys  Pro Ile Arg 
    1175                 1180                 1185             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1190                 1195                 1200             


Arg Lys  Asp Phe Gln Leu Tyr  Lys Val Arg Asp Ile  Asn Asn Tyr 
    1205                 1210                 1215             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1220                 1225                 1230             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1235                 1240                 1245             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1250                 1255                 1260             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Arg Phe Phe  Tyr Ser Asn 
    1265                 1270                 1275             


Ile Met  Asn Phe Phe Lys Thr  Glu Val Lys Leu Ala  Asn Gly Glu 
    1280                 1285                 1290             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1295                 1300                 1305             


Val Val  Trp Asn Lys Glu Lys  Asp Phe Ala Thr Val  Arg Lys Val 
    1310                 1315                 1320             


Leu Ala  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1325                 1330                 1335             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Ser Lys  Arg Glu Ser 
    1340                 1345                 1350             


Ala Lys  Leu Ile Pro Arg Lys  Lys Gly Trp Asp Thr  Arg Lys Tyr 
    1355                 1360                 1365             


Gly Gly  Phe Gly Ser Pro Thr  Val Ala Tyr Ser Ile  Leu Val Val 
    1370                 1375                 1380             


Ala Lys  Val Glu Lys Gly Lys  Ala Lys Lys Leu Lys  Ser Val Lys 
    1385                 1390                 1395             


Val Leu  Val Gly Ile Thr Ile  Met Glu Lys Gly Ser  Tyr Glu Lys 
    1400                 1405                 1410             


Asp Pro  Ile Gly Phe Leu Glu  Ala Lys Gly Tyr Lys  Asp Ile Lys 
    1415                 1420                 1425             


Lys Glu  Leu Ile Phe Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1430                 1435                 1440             


Glu Asn  Gly Arg Arg Arg Met  Leu Ala Ser Ala Thr  Glu Leu Gln 
    1445                 1450                 1455             


Lys Ala  Asn Glu Leu Val Leu  Pro Gln His Leu Val  Arg Leu Leu 
    1460                 1465                 1470             


Tyr Tyr  Thr Gln Asn Ile Ser  Ala Thr Thr Gly Ser  Asn Asn Leu 
    1475                 1480                 1485             


Gly Tyr  Ile Glu Gln His Arg  Glu Glu Phe Lys Glu  Ile Phe Glu 
    1490                 1495                 1500             


Lys Ile  Ile Asp Phe Ser Glu  Lys Tyr Ile Leu Lys  Asn Lys Val 
    1505                 1510                 1515             


Asn Ser  Asn Leu Lys Ser Ser  Phe Asp Glu Gln Phe  Ala Val Ser 
    1520                 1525                 1530             


Asp Ser  Ile Leu Leu Ser Asn  Ser Phe Val Ser Leu  Leu Lys Tyr 
    1535                 1540                 1545             


Thr Ser  Phe Gly Ala Ser Gly  Gly Phe Thr Phe Leu  Asp Leu Asp 
    1550                 1555                 1560             


Val Lys  Gln Gly Arg Leu Arg  Tyr Gln Thr Val Thr  Glu Val Leu 
    1565                 1570                 1575             


Asp Ala  Thr Leu Ile Tyr Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr 
    1580                 1585                 1590             


Arg Thr  Asp Leu Ser Gln Leu  Gly Gly Asp Ser Gly  Gly Ser Lys 
    1595                 1600                 1605             


Arg Thr  Ala Asp Gly Ser Glu  Phe Glu Ser Pro Lys  Lys Lys Arg 
    1610                 1615                 1620             


Lys Val  His His His His His  His 
    1625                 1630     


<210>  491
<211>  1513
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI enhanced, SV40+bipartiteSV40, ancestral mutations 
       in RuvC+REC1/2 domain, HNH deletion longer linker

<400>  491

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val 
    1505                 1510             


<210>  492
<211>  1519
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dCasFok, FokI enhanced SV40+bipartiteSV40, ancestral mutations in
       RuvC+REC1/2 domain, HNH deletion longer linker, 6His tag

<400>  492

Met Ala Val Thr Pro Lys Lys Lys Arg Lys Val Gly Gly Ser Gly Ser 
1               5                   10                  15      


Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
            20                  25                  30          


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
        35                  40                  45              


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
    50                  55                  60                  


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
65                  70                  75                  80  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
                85                  90                  95      


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
            100                 105                 110         


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
        115                 120                 125             


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
    130                 135                 140                 


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
145                 150                 155                 160 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
                165                 170                 175     


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
            180                 185                 190         


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
        195                 200                 205             


Glu Ile Asn Phe Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala 
    210                 215                 220                 


Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
225                 230                 235                 240 


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro 
                245                 250                 255     


Ser Lys Lys Met Lys Val Leu Gly Asn Thr Asp Arg Gln Ser Ile Lys 
            260                 265                 270         


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
        275                 280                 285             


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
    290                 295                 300                 


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ala Asn Glu Met Ala Lys 
305                 310                 315                 320 


Leu Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
                325                 330                 335     


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Leu Val Asp 
            340                 345                 350         


Glu Val Ala Tyr His Glu Asn Tyr Pro Thr Ile Tyr His Leu Arg Lys 
        355                 360                 365             


Lys Leu Ala Asp Ser Thr Glu Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
    370                 375                 380                 


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
385                 390                 395                 400 


Asp Leu Asn Ala Glu Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu 
                405                 410                 415     


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Glu Ser 
            420                 425                 430         


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
        435                 440                 445             


Arg Leu Glu Asn Leu Ile Ala Gln Phe Pro Gly Glu Lys Lys Asn Gly 
    450                 455                 460                 


Leu Phe Gly Asn Ile Ile Ala Leu Ala Leu Gly Leu Thr Pro Asn Phe 
465                 470                 475                 480 


Lys Ser Asn Phe Asp Leu Thr Glu Asp Ala Lys Leu Gln Leu Ser Lys 
                485                 490                 495     


Asp Thr Tyr Asp Asp Asp Leu Asp Glu Leu Leu Gly Gln Ile Gly Asp 
            500                 505                 510         


Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile 
        515                 520                 525             


Leu Leu Ser Asp Ile Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro 
    530                 535                 540                 


Leu Ser Ala Ser Met Val Lys Arg Tyr Asp Glu His His Gln Asp Leu 
545                 550                 555                 560 


Ala Leu Leu Lys Thr Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
                565                 570                 575     


Glu Ile Phe Ser Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
            580                 585                 590         


Ile Gly Ile Lys His Arg Lys Arg Thr Thr Lys Arg Thr Ser Gln Glu 
        595                 600                 605             


Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Ala 
    610                 615                 620                 


Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln 
625                 630                 635                 640 


Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu 
                645                 650                 655     


Leu His Ala Ile Leu Arg Arg Gln Glu Glu Phe Tyr Pro Phe Leu Lys 
            660                 665                 670         


Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr 
        675                 680                 685             


Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr 
    690                 695                 700                 


Arg Lys Ser Asp Glu Ala Ile Thr Pro Trp Asn Phe Glu Glu Val Val 
705                 710                 715                 720 


Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe 
                725                 730                 735     


Asp Glu Tyr Leu Pro Asn Lys Lys Val Leu Pro Lys His Ser Leu Leu 
            740                 745                 750         


Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val 
        755                 760                 765             


Thr Glu Gly Met Arg Lys Pro Glu Phe Leu Ser Gly Glu Gln Lys Glu 
    770                 775                 780                 


Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys 
785                 790                 795                 800 


Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val 
                805                 810                 815     


Glu Ile Ile Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr 
            820                 825                 830         


His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu 
        835                 840                 845             


Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe 
    850                 855                 860                 


Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Lys Tyr Ala Asn Leu 
865                 870                 875                 880 


Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr Gly 
                885                 890                 895     


Trp Gly Arg Leu Ser Arg Lys Met Ile Asn Gly Ile Arg Asp Lys Gln 
            900                 905                 910         


Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ser Asn 
        915                 920                 925             


Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu 
    930                 935                 940                 


Glu Ile Glu Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu 
945                 950                 955                 960 


Gln Ile Ala Asp Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu 
                965                 970                 975     


Gln Thr Val Lys Ile Val Asp Glu Leu Val Lys Val Met Gly His Lys 
            980                 985                 990         


Pro Glu Asn Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Thr 
        995                 1000                 1005             


Lys Gly  Leu Gln Gln Ser Arg  Glu Arg Lys Lys Arg  Gly Gly Ser 
    1010                 1015                 1020             


Ala Gly  Gly Thr Gly Gly Ser  Gly Gly Leu Ser Glu  Ala Asp Lys 
    1025                 1030                 1035             


Ala Gly  Phe Ile Lys Arg Gln  Leu Val Glu Thr Arg  Gln Ile Thr 
    1040                 1045                 1050             


Lys His  Val Ala Arg Ile Leu  Asp Ser Arg Met Asn  Thr Lys Arg 
    1055                 1060                 1065             


Asp Lys  Asn Asp Lys Pro Ile  Arg Glu Val Lys Val  Ile Thr Leu 
    1070                 1075                 1080             


Lys Ser  Lys Leu Val Ser Asp  Phe Arg Lys Asp Phe  Gln Leu Tyr 
    1085                 1090                 1095             


Lys Val  Arg Asp Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
    1100                 1105                 1110             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1115                 1120                 1125             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1130                 1135                 1140             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1145                 1150                 1155             


Ala Lys  Arg Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1160                 1165                 1170             


Glu Val  Lys Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1175                 1180                 1185             


Glu Thr  Asn Gly Glu Thr Gly  Glu Val Val Trp Asn  Lys Glu Lys 
    1190                 1195                 1200             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ala Met Pro  Gln Val Asn 
    1205                 1210                 1215             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1220                 1225                 1230             


Ser Ile  Leu Ser Lys Arg Glu  Ser Ala Lys Leu Ile  Pro Arg Lys 
    1235                 1240                 1245             


Lys Gly  Trp Asp Thr Arg Lys  Tyr Gly Gly Phe Gly  Ser Pro Thr 
    1250                 1255                 1260             


Val Ala  Tyr Ser Ile Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1265                 1270                 1275             


Ala Lys  Lys Leu Lys Ser Val  Lys Val Leu Val Gly  Ile Thr Ile 
    1280                 1285                 1290             


Met Glu  Lys Gly Ser Tyr Glu  Lys Asp Pro Ile Gly  Phe Leu Glu 
    1295                 1300                 1305             


Ala Lys  Gly Tyr Lys Asp Ile  Lys Lys Glu Leu Ile  Phe Lys Leu 
    1310                 1315                 1320             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Arg Arg Met 
    1325                 1330                 1335             


Leu Ala  Ser Ala Thr Glu Leu  Gln Lys Ala Asn Glu  Leu Val Leu 
    1340                 1345                 1350             


Pro Gln  His Leu Val Arg Leu  Leu Tyr Tyr Thr Gln  Asn Ile Ser 
    1355                 1360                 1365             


Ala Thr  Thr Gly Ser Asn Asn  Leu Gly Tyr Ile Glu  Gln His Arg 
    1370                 1375                 1380             


Glu Glu  Phe Lys Glu Ile Phe  Glu Lys Ile Ile Asp  Phe Ser Glu 
    1385                 1390                 1395             


Lys Tyr  Ile Leu Lys Asn Lys  Val Asn Ser Asn Leu  Lys Ser Ser 
    1400                 1405                 1410             


Phe Asp  Glu Gln Phe Ala Val  Ser Asp Ser Ile Leu  Leu Ser Asn 
    1415                 1420                 1425             


Ser Phe  Val Ser Leu Leu Lys  Tyr Thr Ser Phe Gly  Ala Ser Gly 
    1430                 1435                 1440             


Gly Phe  Thr Phe Leu Asp Leu  Asp Val Lys Gln Gly  Arg Leu Arg 
    1445                 1450                 1455             


Tyr Gln  Thr Val Thr Glu Val  Leu Asp Ala Thr Leu  Ile Tyr Gln 
    1460                 1465                 1470             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Thr Asp Leu  Ser Gln Leu 
    1475                 1480                 1485             


Gly Gly  Asp Ser Gly Gly Ser  Lys Arg Thr Ala Asp  Gly Ser Glu 
    1490                 1495                 1500             


Phe Glu  Ser Pro Lys Lys Lys  Arg Lys Val His His  His His His 
    1505                 1510                 1515             


His 
    


<210>  493
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CTLA4 gene 5'-3' fragment of figure 9A, set 1

<400>  493
cttgccttgg atttcagcgg cacaaggctc agctgaacct ggctaccagg acctggccct       60

g                                                                       61


<210>  494
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CTLA4 gene 3'-5' fragment of figure 9A, set 1

<400>  494
gaacggaacc taaagtcgcc gtgttccgag tcgacttgga ccgatggtcc tggaccggga       60

c                                                                       61


<210>  495
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CTLA4 gene 5'-3' fragment of figure 9B, set 2

<400>  495
ctgaacctgg ctaccaggac ctggccctgc actctcctgt tttttcttct cttcatccct       60

g                                                                       61


<210>  496
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CTLA4 gene 3'-5' fragment of figure 9B, set 2

<400>  496
gacttggacc gatggtcctg gaccgggacg tgagaggaca aaaaagaaga gaagtaggga       60

c                                                                       61


<210>  497
<211>  77
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SIGLEC10 gene 5'-3' fragment of figure 10

<400>  497
ccctgcgcct cctatgcgga gatgctactg ccactgctgc tgtcctcgct gctgggcggt       60

gagtgggcca agggccg                                                      77


<210>  498
<211>  77
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SIGLEC10gene 3'-5' fragment of figure 10

<400>  498
gggacgcgga ggatacgcct ctacgatgac ggtgacgacg acaggagcga cgacccgcca       60

ctcacccggt tcccggc                                                      77


<210>  499
<211>  80
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ITGB3 gene 5'-3' fragment of figure 11

<400>  499
cgagcgcggc ggctctgggc agcggtgctg gtggtggggg cgctggcggg cgtcggcgtc       60

ggaggtgagt ggggctcccg                                                   80


<210>  500
<211>  80
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ITGB3gene 3'-5' fragment of figure 11

<400>  500
gctcgcgccg ccgagacccg tcgccacgac caccaccccc gcgaccgccc gcagccgcag       60

cctccactca ccccgagggc                                                   80


<210>  501
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CTLA4 gene 5'-3' fragment of figure 12

<400>  501
ttcctgaaga cctgaacacc gctcccataa agccatggct tgccttggat ttcagcggca       60

caa                                                                     63


<210>  502
<211>  63
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CTLA4gene 3'-5' fragment of figure 12

<400>  502
aaggacttct ggacttgtgg cgagggtatt tcggtaccga acggaaccta aagtcgccgt       60

gtt                                                                     63


<210>  503
<211>  268
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PAM binding C-terminal domain 1108-1375 of scCas9

<400>  503

Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Ser Lys Arg 
1               5                   10                  15      


Glu Ser Ala Lys Leu Ile Pro Arg Lys Lys Gly Trp Asp Thr Arg Lys 
            20                  25                  30          


Tyr Gly Gly Phe Gly Ser Pro Thr Val Ala Tyr Ser Ile Leu Val Val 
        35                  40                  45              


Ala Lys Val Glu Lys Gly Lys Ala Lys Lys Leu Lys Ser Val Lys Val 
    50                  55                  60                  


Leu Val Gly Ile Thr Ile Met Glu Lys Gly Ser Tyr Glu Lys Asp Pro 
65                  70                  75                  80  


Ile Gly Phe Leu Glu Ala Lys Gly Tyr Lys Asp Ile Lys Lys Glu Leu 
                85                  90                  95      


Ile Phe Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg 
            100                 105                 110         


Arg Arg Met Leu Ala Ser Ala Thr Glu Leu Gln Lys Ala Asn Glu Leu 
        115                 120                 125             


Val Leu Pro Gln His Leu Val Arg Leu Leu Tyr Tyr Thr Gln Asn Ile 
    130                 135                 140                 


Ser Ala Thr Thr Gly Ser Asn Asn Leu Gly Tyr Ile Glu Gln His Arg 
145                 150                 155                 160 


Glu Glu Phe Lys Glu Ile Phe Glu Lys Ile Ile Asp Phe Ser Glu Lys 
                165                 170                 175     


Tyr Ile Leu Lys Asn Lys Val Asn Ser Asn Leu Lys Ser Ser Phe Asp 
            180                 185                 190         


Glu Gln Phe Ala Val Ser Asp Ser Ile Leu Leu Ser Asn Ser Phe Val 
        195                 200                 205             


Ser Leu Leu Lys Tyr Thr Ser Phe Gly Ala Ser Gly Gly Phe Thr Phe 
    210                 215                 220                 


Leu Asp Leu Asp Val Lys Gln Gly Arg Leu Arg Tyr Gln Thr Val Thr 
225                 230                 235                 240 


Glu Val Leu Asp Ala Thr Leu Ile Tyr Gln Ser Ile Thr Gly Leu Tyr 
                245                 250                 255     


Glu Thr Arg Thr Asp Leu Ser Gln Leu Gly Gly Asp 
            260                 265             


<210>  504
<211>  116
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PAMBD: 1228-343

<400>  504

Glu Leu Gln Lys Ala Asn Glu Leu Val Leu Pro Gln His Leu Val Arg 
1               5                   10                  15      


Leu Leu Tyr Tyr Thr Gln Asn Ile Ser Ala Thr Thr Gly Ser Asn Asn 
            20                  25                  30          


Leu Gly Tyr Ile Glu Gln His Arg Glu Glu Phe Lys Glu Ile Phe Glu 
        35                  40                  45              


Lys Ile Ile Asp Phe Ser Glu Lys Tyr Ile Leu Lys Asn Lys Val Asn 
    50                  55                  60                  


Ser Asn Leu Lys Ser Ser Phe Asp Glu Gln Phe Ala Val Ser Asp Ser 
65                  70                  75                  80  


Ile Leu Leu Ser Asn Ser Phe Val Ser Leu Leu Lys Tyr Thr Ser Phe 
                85                  90                  95      


Gly Ala Ser Gly Gly Phe Thr Phe Leu Asp Leu Asp Val Lys Gln Gly 
            100                 105                 110         


Arg Leu Arg Tyr 
        115     


<210>  505
<211>  13
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PAMBD LOOP 1330-1342

<400>  505

Thr Phe Leu Asp Leu Asp Val Lys Gln Gly Arg Leu Arg 
1               5                   10              


<210>  506
<211>  122
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  ScCas9 HNH domain, residues 793-914 of ScCas9

<400>  506

Ile Glu Glu Gly Ile Lys Glu Leu Glu Ser Gln Ile Leu Lys Glu Asn 
1               5                   10                  15      


Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr 
            20                  25                  30          


Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn 
        35                  40                  45              


Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Ile 
    50                  55                  60                  


Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Val Glu Asn 
65                  70                  75                  80  


Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met 
                85                  90                  95      


Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg 
            100                 105                 110         


Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
        115                 120         


