                         SEQUENCE LISTING

<110>  Deutsches Krebsforschungszentrum
       Universitt Heidelberg
       Berliner Institut fr Gesundheitsforschung Institute of Health  BIH
 
<120>  Improving Cas Nuclease Target Specificity

<130>  DK15992PC

<150>  EP19186590.6
<151>  2019-07-16

<160>  65    

<170>  PatentIn version 3.5

<210>  1
<211>  1367
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Cas9

<400>  1

Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val Gly 
1               5                   10                  15      


Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys 
            20                  25                  30          


Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly 
        35                  40                  45              


Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys 
    50                  55                  60                  


Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr 
65                  70                  75                  80  


Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe 
                85                  90                  95      


Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His 
            100                 105                 110         


Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His 
        115                 120                 125             


Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser 
    130                 135                 140                 


Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met 
145                 150                 155                 160 


Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp 
                165                 170                 175     


Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn 
            180                 185                 190         


Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys 
        195                 200                 205             


Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu 
    210                 215                 220                 


Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu 
225                 230                 235                 240 


Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp 
                245                 250                 255     


Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp 
            260                 265                 270         


Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu 
        275                 280                 285             


Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile 
    290                 295                 300                 


Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met 
305                 310                 315                 320 


Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala 
                325                 330                 335     


Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp 
            340                 345                 350         


Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln 
        355                 360                 365             


Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
    370                 375                 380                 


Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys 
385                 390                 395                 400 


Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly 
                405                 410                 415     


Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu 
            420                 425                 430         


Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
        435                 440                 445             


Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met 
    450                 455                 460                 


Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val 
465                 470                 475                 480 


Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
                485                 490                 495     


Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu 
            500                 505                 510         


Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
        515                 520                 525             


Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys 
    530                 535                 540                 


Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
545                 550                 555                 560 


Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
                565                 570                 575     


Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
            580                 585                 590         


Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
        595                 600                 605             


Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
    610                 615                 620                 


Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
625                 630                 635                 640 


Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr 
                645                 650                 655     


Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys 
            660                 665                 670         


Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala 
        675                 680                 685             


Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys 
    690                 695                 700                 


Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His 
705                 710                 715                 720 


Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile 
                725                 730                 735     


Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg 
            740                 745                 750         


His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr 
        755                 760                 765             


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


Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val 
785                 790                 795                 800 


Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln 
                805                 810                 815     


Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu 
            820                 825                 830         


Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys Asp 
        835                 840                 845             


Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly 
    850                 855                 860                 


Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn 
865                 870                 875                 880 


Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe 
                885                 890                 895     


Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys 
            900                 905                 910         


Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys 
        915                 920                 925             


His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu 
    930                 935                 940                 


Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys 
945                 950                 955                 960 


Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu 
                965                 970                 975     


Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val 
            980                 985                 990         


Gly Thr Ala Leu Ile Lys Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val 
        995                 1000                 1005             


Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys 
    1010                 1015                 1020             


Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr 
    1025                 1030                 1035             


Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn 
    1040                 1045                 1050             


Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr 
    1055                 1060                 1065             


Gly Glu  Ile Val Trp Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg 
    1070                 1075                 1080             


Lys Val  Leu Ser Met Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu 
    1085                 1090                 1095             


Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg 
    1100                 1105                 1110             


Asn Ser  Asp Lys Leu Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys 
    1115                 1120                 1125             


Lys Tyr  Gly Gly Phe Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu 
    1130                 1135                 1140             


Val Val  Ala Lys Val Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser 
    1145                 1150                 1155             


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


Glu Lys  Asn Pro Ile Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu 
    1175                 1180                 1185             


Val Lys  Lys Asp Leu Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe 
    1190                 1195                 1200             


Glu Leu  Glu Asn Gly Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu 
    1205                 1210                 1215             


Leu Gln  Lys Gly Asn Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn 
    1220                 1225                 1230             


Phe Leu  Tyr Leu Ala Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro 
    1235                 1240                 1245             


Glu Asp  Asn Glu Gln Lys Gln  Leu Phe Val Glu Gln  His Lys His 
    1250                 1255                 1260             


Tyr Leu  Asp Glu Ile Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg 
    1265                 1270                 1275             


Val Ile  Leu Ala Asp Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr 
    1280                 1285                 1290             


Asn Lys  His Arg Asp Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile 
    1295                 1300                 1305             


Ile His  Leu Phe Thr Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe 
    1310                 1315                 1320             


Lys Tyr  Phe Asp Thr Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr 
    1325                 1330                 1335             


Lys Glu  Val Leu Asp Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly 
    1340                 1345                 1350             


Leu Tyr  Glu Thr Arg Ile Asp  Leu Ser Gln Leu Gly  Gly Asp 
    1355                 1360                 1365         


<210>  2
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Cas9 encoding

<400>  2
gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc      180

acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat      240

ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg      300

gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg      660

ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg      720

attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat      780

gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg      900

ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg      960

atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag     1020

cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa     1140

aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc     1260

attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag     1320

aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga     1380

ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg     1440

gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat     1560

aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc     1620

ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg     1680

aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc     1740

ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc     1800

aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg     1860

accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac     1920

ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat     2040

ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac     2160

gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg     2220

aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg     2340

aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg     2400

gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat     2460

atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc     2520

gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac     2580

aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc     2700

aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg     2760

gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact     2820

aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag     2880

ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac     3000

cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac     3120

atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct     3180

ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc     3240

accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag     3300

acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc     3360

agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat     3420

tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa     3480

gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt     3540

ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac     3600

tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag     3660

aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac     3720

tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc     3840

ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc     3900

atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct     3960

gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac     4080

ctgtctcagc tgggaggcga c                                               4101


<210>  3
<211>  87
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  AcrIIA4

<400>  3

Met Asn Ile Asn Asp Leu Ile Arg Glu Ile Lys Asn Lys Asp Tyr Thr 
1               5                   10                  15      


Val Lys Leu Ser Gly Thr Asp Ser Asn Ser Ile Thr Gln Leu Ile Ile 
            20                  25                  30          


Arg Val Asn Asn Asp Gly Asn Glu Tyr Val Ile Ser Glu Ser Glu Asn 
        35                  40                  45              


Glu Ser Ile Val Glu Lys Phe Ile Ser Ala Phe Lys Asn Gly Trp Asn 
    50                  55                  60                  


Gln Glu Tyr Glu Asp Glu Glu Glu Phe Tyr Asn Asp Met Gln Thr Ile 
65                  70                  75                  80  


Thr Leu Lys Ser Glu Leu Asn 
                85          


<210>  4
<211>  261
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AcrIIA4 encoding

<400>  4
atgaacatta acgacctgat tcgggaaata aagaacaagg actacacggt caagctgtca       60

ggcaccgact ccaactctat aacacagctc attataaggg ttaataatga tggcaacgaa      120

tatgtcatct ctgaatccga gaatgaatca atcgtggaga aattcatcag tgcatttaaa      180

aacggctgga accaggagta cgaagatgag gaagagttct acaatgatat gcaaactatc      240

acactgaaat ctgagctgaa c                                                261


<210>  5
<211>  87
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  AcrIIA4 N39A

<400>  5

Met Asn Ile Asn Asp Leu Ile Arg Glu Ile Lys Asn Lys Asp Tyr Thr 
1               5                   10                  15      


Val Lys Leu Ser Gly Thr Asp Ser Asn Ser Ile Thr Gln Leu Ile Ile 
            20                  25                  30          


Arg Val Asn Asn Asp Gly Ala Glu Tyr Val Ile Ser Glu Ser Glu Asn 
        35                  40                  45              


Glu Ser Ile Val Glu Lys Phe Ile Ser Ala Phe Lys Asn Gly Trp Asn 
    50                  55                  60                  


Gln Glu Tyr Glu Asp Glu Glu Glu Phe Tyr Asn Asp Met Gln Thr Ile 
65                  70                  75                  80  


Thr Leu Lys Ser Glu Leu Asn 
                85          


<210>  6
<211>  261
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AcrIIA4 N39A encoding

<400>  6
atgaacatta acgacctgat tcgggaaata aagaacaagg actacacggt caagctgtca       60

ggcaccgact ccaactctat aacacagctc attataaggg ttaataatga tggcgccgaa      120

tatgtcatct ctgaatccga gaatgaatca atcgtggaga aattcatcag tgcatttaaa      180

aacggctgga accaggagta cgaagatgag gaagagttct acaatgatat gcaaactatc      240

acactgaaat ctgagctgaa c                                                261


<210>  7
<211>  87
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  AcrIIA4 D14A/G38A

<400>  7

Met Asn Ile Asn Asp Leu Ile Arg Glu Ile Lys Asn Lys Ala Tyr Thr 
1               5                   10                  15      


Val Lys Leu Ser Gly Thr Asp Ser Asn Ser Ile Thr Gln Leu Ile Ile 
            20                  25                  30          


Arg Val Asn Asn Asp Ala Asn Glu Tyr Val Ile Ser Glu Ser Glu Asn 
        35                  40                  45              


Glu Ser Ile Val Glu Lys Phe Ile Ser Ala Phe Lys Asn Gly Trp Asn 
    50                  55                  60                  


Gln Glu Tyr Glu Asp Glu Glu Glu Phe Tyr Asn Asp Met Gln Thr Ile 
65                  70                  75                  80  


Thr Leu Lys Ser Glu Leu Asn 
                85          


<210>  8
<211>  261
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AcrIIA4 D14A/G38A encoding

<400>  8
atgaacatta acgacctgat tcgggaaata aagaacaagg cctacacggt caagctgtca       60

ggcaccgact ccaactctat aacacagctc attataaggg ttaataatga tgccaacgaa      120

tatgtcatct ctgaatccga gaatgaatca atcgtggaga aattcatcag tgcatttaaa      180

aacggctgga accaggagta cgaagatgag gaagagttct acaatgatat gcaaactatc      240

acactgaaat ctgagctgaa c                                                261


<210>  9
<211>  227
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  AcrIIA4 del N64/Q65/E66 + LOV

<400>  9

Met Asn Ile Asn Asp Leu Ile Arg Glu Ile Lys Asn Lys Asp Tyr Thr 
1               5                   10                  15      


Val Lys Leu Ser Gly Thr Asp Ser Asn Ser Ile Thr Gln Leu Ile Ile 
            20                  25                  30          


Arg Val Asn Asn Asp Gly Asn Glu Tyr Val Ile Ser Glu Ser Glu Asn 
        35                  40                  45              


Glu Ser Ile Val Glu Lys Phe Ile Ser Ala Phe Lys Asn Gly Trp Leu 
    50                  55                  60                  


Ala Ala Ala Leu Glu Arg Ile Glu Lys Asn Phe Val Ile Thr Asp Pro 
65                  70                  75                  80  


Arg Leu Pro Asp Asn Pro Ile Ile Phe Ala Ser Asp Ser Phe Leu Gln 
                85                  90                  95      


Leu Thr Glu Tyr Ser Arg Glu Glu Ile Leu Gly Arg Asn Cys Arg Phe 
            100                 105                 110         


Leu Gln Gly Pro Glu Thr Asp Arg Ala Thr Val Arg Lys Ile Arg Asp 
        115                 120                 125             


Ala Ile Asp Asn Gln Thr Glu Val Thr Val Gln Leu Ile Asn Tyr Thr 
    130                 135                 140                 


Lys Ser Gly Lys Lys Phe Trp Asn Leu Phe His Leu Gln Pro Met Arg 
145                 150                 155                 160 


Asp Gln Lys Gly Asp Val Gln Tyr Phe Ile Gly Val Gln Leu Asp Gly 
                165                 170                 175     


Thr Glu His Val Arg Asp Ala Ala Glu Arg Glu Ala Val Met Leu Ile 
            180                 185                 190         


Lys Lys Thr Ala Glu Glu Ile Asp Glu Ala Ala Lys Glu Leu Tyr Glu 
        195                 200                 205             


Asp Glu Glu Glu Phe Tyr Asn Asp Met Gln Thr Ile Thr Leu Lys Ser 
    210                 215                 220                 


Glu Leu Asn 
225         


<210>  10
<211>  681
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AcrIIA4 del N64/Q65/E66 + LOV encoding

<400>  10
atgaacatta acgacctgat tcgggaaata aagaacaagg actacacggt caagctgtca       60

ggcaccgact ccaactctat aacacagctc attataaggg ttaataatga tggcaacgaa      120

tatgtcatct ctgaatccga gaatgaatca atcgtggaga aattcatcag tgcatttaaa      180

aacggctggc ttgctgcagc gttggagcgg attgagaaga atttcgtcat aactgatccc      240

aggttgcctg ataatcctat aatattcgcc tccgatagtt ttcttcaact tacggagtac      300

tccagagagg agatactggg aaggaattgc agattcctcc aaggtccaga aactgataga      360

gctactgtga ggaagatacg ggatgccatt gacaatcaaa ctgaagtgac agtgcaactt      420

atcaattaca ccaagtcagg gaaaaaattc tggaacctgt tccatttgca acccatgagg      480

gaccaaaagg gcgatgtgca atatttcata ggcgttcaac ttgatggtac tgaacacgtc      540

agagatgcgg cggagcgaga agcagtgatg ctgattaaga aaacagcaga ggagatcgac      600

gaagcggcga aagagttgta cgaagatgag gaagagttct acaatgatat gcaaactatc      660

acactgaaat ctgagctgaa c                                                681


<210>  11
<211>  143
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  LOV2 domain

<400>  11

Leu Ala Thr Thr Leu Glu Arg Ile Glu Lys Asn Phe Val Ile Thr Asp 
1               5                   10                  15      


Pro Arg Leu Pro Asp Asn Pro Ile Ile Phe Ala Ser Asp Ser Phe Leu 
            20                  25                  30          


Gln Leu Thr Glu Tyr Ser Arg Glu Glu Ile Leu Gly Arg Asn Cys Arg 
        35                  40                  45              


Phe Leu Gln Gly Pro Glu Thr Asp Arg Ala Thr Val Arg Lys Ile Arg 
    50                  55                  60                  


Asp Ala Ile Asp Asn Gln Thr Glu Val Thr Val Gln Leu Ile Asn Tyr 
65                  70                  75                  80  


Thr Lys Ser Gly Lys Lys Phe Trp Asn Leu Phe His Leu Gln Pro Met 
                85                  90                  95      


Arg Asp Gln Lys Gly Asp Val Gln Tyr Phe Ile Gly Val Gln Leu Asp 
            100                 105                 110         


Gly Thr Glu His Val Arg Asp Ala Ala Glu Arg Glu Gly Val Met Leu 
        115                 120                 125             


Ile Lys Lys Thr Ala Glu Asn Ile Asp Glu Ala Ala Lys Glu Leu 
    130                 135                 140             


<210>  12
<211>  429
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  LOV2 domain encoding

<400>  12
cttgctacaa ctttggagcg gattgagaag aatttcgtca taactgatcc caggttgcct       60

gataatccta taatattcgc ctccgatagt tttcttcaac ttacggagta ctccagagag      120

gagatactgg gaaggaattg cagattcctc caaggtccag aaactgatag agctactgtg      180

aggaagatac gggatgccat tgacaatcaa actgaagtga cagtgcaact tatcaattac      240

accaagtcag ggaaaaaatt ctggaacctg ttccatttgc aacccatgag ggaccaaaag      300

ggcgatgtgc aatatttcat aggcgttcaa cttgatggta ctgaacacgt cagagatgcg      360

gcggagcgag aaggagtgat gctgattaag aaaacagcag agaatatcga cgaagcggcg      420

aaagagttg                                                              429


<210>  13
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  GGSG linker

<400>  13

Gly Gly Ser Gly 
1               


<210>  14
<211>  40
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  GGSG linker long

<400>  14

Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly 
1               5                   10                  15      


Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly 
            20                  25                  30          


Gly Gly Ser Gly Gly Gly Ser Gly 
        35                  40  


<210>  15
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#1 (NLS-Cas9-NLS fused to wild-type AcrIIA4)

<400>  15

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  16
<211>  1692
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#2 (NLS-Cas9-NLS fused to AcrIIA4 with N64/Q65/E66 
       deletion and an insertion of a LOV-domain)

<400>  16

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Leu Ala  Thr Thr Leu 
    1520                 1525                 1530             


Glu Arg  Ile Glu Lys Asn Phe  Val Ile Thr Asp Pro  Arg Leu Pro 
    1535                 1540                 1545             


Asp Asn  Pro Ile Ile Phe Ala  Ser Asp Ser Phe Leu  Gln Leu Thr 
    1550                 1555                 1560             


Glu Tyr  Ser Arg Glu Glu Ile  Leu Gly Arg Asn Cys  Arg Phe Leu 
    1565                 1570                 1575             


Gln Gly  Pro Glu Thr Asp Arg  Ala Thr Val Arg Lys  Ile Arg Asp 
    1580                 1585                 1590             


Ala Ile  Asp Asn Gln Thr Glu  Val Thr Val Gln Leu  Ile Asn Tyr 
    1595                 1600                 1605             


Thr Lys  Ser Gly Lys Lys Phe  Trp Asn Leu Phe His  Leu Gln Pro 
    1610                 1615                 1620             


Met Arg  Asp Gln Lys Gly Asp  Val Gln Tyr Phe Ile  Gly Val Gln 
    1625                 1630                 1635             


Leu Asp  Gly Thr Glu His Val  Arg Asp Ala Ala Glu  Arg Glu Gly 
    1640                 1645                 1650             


Val Met  Leu Ile Lys Lys Thr  Ala Glu Asn Ile Asp  Glu Ala Ala 
    1655                 1660                 1665             


Lys Glu  Leu Tyr Glu Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln 
    1670                 1675                 1680             


Thr Ile  Thr Leu Lys Ser Glu  Leu Asn 
    1685                 1690         


<210>  17
<211>  1695
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#3 (NLS-Cas9-NLS fused to AcrIIA4 with Q65/E66 deletion 
       and an insertion of a LOV-domain flanked by glycine linker 
       residues)

<400>  17

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gly  Leu Ala Thr 
    1520                 1525                 1530             


Thr Leu  Glu Arg Ile Glu Lys  Asn Phe Val Ile Thr  Asp Pro Arg 
    1535                 1540                 1545             


Leu Pro  Asp Asn Pro Ile Ile  Phe Ala Ser Asp Ser  Phe Leu Gln 
    1550                 1555                 1560             


Leu Thr  Glu Tyr Ser Arg Glu  Glu Ile Leu Gly Arg  Asn Cys Arg 
    1565                 1570                 1575             


Phe Leu  Gln Gly Pro Glu Thr  Asp Arg Ala Thr Val  Arg Lys Ile 
    1580                 1585                 1590             


Arg Asp  Ala Ile Asp Asn Gln  Thr Glu Val Thr Val  Gln Leu Ile 
    1595                 1600                 1605             


Asn Tyr  Thr Lys Ser Gly Lys  Lys Phe Trp Asn Leu  Phe His Leu 
    1610                 1615                 1620             


Gln Pro  Met Arg Asp Gln Lys  Gly Asp Val Gln Tyr  Phe Ile Gly 
    1625                 1630                 1635             


Val Gln  Leu Asp Gly Thr Glu  His Val Arg Asp Ala  Ala Glu Arg 
    1640                 1645                 1650             


Glu Gly  Val Met Leu Ile Lys  Lys Thr Ala Glu Asn  Ile Asp Glu 
    1655                 1660                 1665             


Ala Ala  Lys Glu Leu Gly Tyr  Glu Asp Glu Glu Glu  Phe Tyr Asn 
    1670                 1675                 1680             


Asp Met  Gln Thr Ile Thr Leu  Lys Ser Glu Leu Asn  
    1685                 1690                 1695 


<210>  18
<211>  1699
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#4 (NLS-Cas9-NLS fused to AcrIIA4 with Q65/E66 deletion 
       and an insertion of a LOV-domain flanked by Gly-Ser-Gly linkers)

<400>  18

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gly  Ser Gly Leu 
    1520                 1525                 1530             


Ala Thr  Thr Leu Glu Arg Ile  Glu Lys Asn Phe Val  Ile Thr Asp 
    1535                 1540                 1545             


Pro Arg  Leu Pro Asp Asn Pro  Ile Ile Phe Ala Ser  Asp Ser Phe 
    1550                 1555                 1560             


Leu Gln  Leu Thr Glu Tyr Ser  Arg Glu Glu Ile Leu  Gly Arg Asn 
    1565                 1570                 1575             


Cys Arg  Phe Leu Gln Gly Pro  Glu Thr Asp Arg Ala  Thr Val Arg 
    1580                 1585                 1590             


Lys Ile  Arg Asp Ala Ile Asp  Asn Gln Thr Glu Val  Thr Val Gln 
    1595                 1600                 1605             


Leu Ile  Asn Tyr Thr Lys Ser  Gly Lys Lys Phe Trp  Asn Leu Phe 
    1610                 1615                 1620             


His Leu  Gln Pro Met Arg Asp  Gln Lys Gly Asp Val  Gln Tyr Phe 
    1625                 1630                 1635             


Ile Gly  Val Gln Leu Asp Gly  Thr Glu His Val Arg  Asp Ala Ala 
    1640                 1645                 1650             


Glu Arg  Glu Gly Val Met Leu  Ile Lys Lys Thr Ala  Glu Asn Ile 
    1655                 1660                 1665             


Asp Glu  Ala Ala Lys Glu Leu  Gly Ser Gly Tyr Glu  Asp Glu Glu 
    1670                 1675                 1680             


Glu Phe  Tyr Asn Asp Met Gln  Thr Ile Thr Leu Lys  Ser Glu Leu 
    1685                 1690                 1695             


Asn 
    


<210>  19
<211>  1692
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#5 (NLS-Cas9-NLS fused to AcrIIA4 with N64/Q65/E66 
       deletion and an insertion of a LOV-domain bearing 
       T406A/T407A/G528A/N538E mutations)

<400>  19

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Leu Ala  Ala Ala Leu 
    1520                 1525                 1530             


Glu Arg  Ile Glu Lys Asn Phe  Val Ile Thr Asp Pro  Arg Leu Pro 
    1535                 1540                 1545             


Asp Asn  Pro Ile Ile Phe Ala  Ser Asp Ser Phe Leu  Gln Leu Thr 
    1550                 1555                 1560             


Glu Tyr  Ser Arg Glu Glu Ile  Leu Gly Arg Asn Cys  Arg Phe Leu 
    1565                 1570                 1575             


Gln Gly  Pro Glu Thr Asp Arg  Ala Thr Val Arg Lys  Ile Arg Asp 
    1580                 1585                 1590             


Ala Ile  Asp Asn Gln Thr Glu  Val Thr Val Gln Leu  Ile Asn Tyr 
    1595                 1600                 1605             


Thr Lys  Ser Gly Lys Lys Phe  Trp Asn Leu Phe His  Leu Gln Pro 
    1610                 1615                 1620             


Met Arg  Asp Gln Lys Gly Asp  Val Gln Tyr Phe Ile  Gly Val Gln 
    1625                 1630                 1635             


Leu Asp  Gly Thr Glu His Val  Arg Asp Ala Ala Glu  Arg Glu Ala 
    1640                 1645                 1650             


Val Met  Leu Ile Lys Lys Thr  Ala Glu Glu Ile Asp  Glu Ala Ala 
    1655                 1660                 1665             


Lys Glu  Leu Tyr Glu Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln 
    1670                 1675                 1680             


Thr Ile  Thr Leu Lys Ser Glu  Leu Asn 
    1685                 1690         


<210>  20
<211>  1692
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#6 (NLS-Cas9-NLS fused to AcrIIA4 with N64/Q65/E66 
       deletion and an insertion of a LOV-domain bearing T406A/T407A 
       mutations)

<400>  20

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Leu Ala  Ala Ala Leu 
    1520                 1525                 1530             


Glu Arg  Ile Glu Lys Asn Phe  Val Ile Thr Asp Pro  Arg Leu Pro 
    1535                 1540                 1545             


Asp Asn  Pro Ile Ile Phe Ala  Ser Asp Ser Phe Leu  Gln Leu Thr 
    1550                 1555                 1560             


Glu Tyr  Ser Arg Glu Glu Ile  Leu Gly Arg Asn Cys  Arg Phe Leu 
    1565                 1570                 1575             


Gln Gly  Pro Glu Thr Asp Arg  Ala Thr Val Arg Lys  Ile Arg Asp 
    1580                 1585                 1590             


Ala Ile  Asp Asn Gln Thr Glu  Val Thr Val Gln Leu  Ile Asn Tyr 
    1595                 1600                 1605             


Thr Lys  Ser Gly Lys Lys Phe  Trp Asn Leu Phe His  Leu Gln Pro 
    1610                 1615                 1620             


Met Arg  Asp Gln Lys Gly Asp  Val Gln Tyr Phe Ile  Gly Val Gln 
    1625                 1630                 1635             


Leu Asp  Gly Thr Glu His Val  Arg Asp Ala Ala Glu  Arg Glu Gly 
    1640                 1645                 1650             


Val Met  Leu Ile Lys Lys Thr  Ala Glu Asn Ile Asp  Glu Ala Ala 
    1655                 1660                 1665             


Lys Glu  Leu Tyr Glu Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln 
    1670                 1675                 1680             


Thr Ile  Thr Leu Lys Ser Glu  Leu Asn 
    1685                 1690         


<210>  21
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#7 (NLS-Cas9-NLS fused to AcrIIA4 bearing a M77A mutation)

<400>  21

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Ala Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  22
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#8 (NLS-Cas9-NLS fused to AcrIIA4 bearing D76A/M77A 
       mutations)

<400>  22

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Ala Ala Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  23
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#9 (NLS-Cas9-NLS fused to AcrIIA4 bearing a D14A mutation)

<400>  23

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Ala Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  24
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#10 (NLS-Cas9-NLS fused to AcrIIA4 bearing D14A/Y15A 
       mutation)

<400>  24

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Ala Ala  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  25
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#11 (NLS-Cas9-NLS fused to AcrIIA4 bearing a N39A 
       mutation)

<400>  25

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Gly 
    1490                 1495                 1500             


Ala Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  26
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#12 (NLS-Cas9-NLS fused to AcrIIA4 bearing a G38A 
       mutation)

<400>  26

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Ala 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  27
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#13 (NLS-Cas9-NLS fused to AcrIIA4 bearing D37A/G38A 
       mutation)

<400>  27

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Asp Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Ala Ala 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  28
<211>  1552
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CasCAID#14 (NLS-Cas9-NLS fused to AcrIIA4 bearing D14A/G38A 
       mutation)

<400>  28

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              


Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             


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


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         


Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 


Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 


Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     


Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         


Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             


Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 


Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 


Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     


His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         


Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             


Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 


Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 


Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     


Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         


Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             


Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 


Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 


Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     


Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         


Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             


Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 


Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 


Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     


Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         


Lys Val Ile Thr Leu Lys Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp 
        995                 1000                 1005             


Phe Gln  Phe Tyr Lys Val Arg  Glu Ile Asn Asn Tyr  His His Ala 
    1010                 1015                 1020             


His Asp  Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
    1025                 1030                 1035             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1040                 1045                 1050             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1055                 1060                 1065             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1070                 1075                 1080             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1085                 1090                 1095             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1100                 1105                 1110             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1115                 1120                 1125             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1130                 1135                 1140             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1145                 1150                 1155             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1160                 1165                 1170             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1175                 1180                 1185             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1190                 1195                 1200             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1205                 1210                 1215             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1220                 1225                 1230             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1235                 1240                 1245             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1250                 1255                 1260             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1265                 1270                 1275             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1280                 1285                 1290             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1295                 1300                 1305             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1310                 1315                 1320             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1325                 1330                 1335             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1340                 1345                 1350             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1355                 1360                 1365             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1370                 1375                 1380             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1385                 1390                 1395             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Lys Arg Pro  Ala Ala Thr 
    1400                 1405                 1410             


Lys Lys  Ala Gly Gln Ala Lys  Lys Lys Lys Glu Phe  Gly Gly Ser 
    1415                 1420                 1425             


Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly Gly Ser  Gly Gly Gly 
    1430                 1435                 1440             


Ser Gly  Gly Gly Ser Gly Gly  Gly Ser Gly Gly Gly  Ser Gly Gly 
    1445                 1450                 1455             


Gly Ser  Gly Gly Gly Ser Gly  Met Asn Ile Asn Asp  Leu Ile Arg 
    1460                 1465                 1470             


Glu Ile  Lys Asn Lys Ala Tyr  Thr Val Lys Leu Ser  Gly Thr Asp 
    1475                 1480                 1485             


Ser Asn  Ser Ile Thr Gln Leu  Ile Ile Arg Val Asn  Asn Asp Ala 
    1490                 1495                 1500             


Asn Glu  Tyr Val Ile Ser Glu  Ser Glu Asn Glu Ser  Ile Val Glu 
    1505                 1510                 1515             


Lys Phe  Ile Ser Ala Phe Lys  Asn Gly Trp Asn Gln  Glu Tyr Glu 
    1520                 1525                 1530             


Asp Glu  Glu Glu Phe Tyr Asn  Asp Met Gln Thr Ile  Thr Leu Lys 
    1535                 1540                 1545             


Ser Glu  Leu Asn 
    1550         


<210>  29
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1 ON

<400>  29
gggagggaga gcttggcagg ggg                                               23


<210>  30
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1 OFF1

<400>  30
gggaagggga gcttggcagg tgg                                               23


<210>  31
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1 OFF2

<400>  31
gggaaggtga gcttggcagg tgg                                               23


<210>  32
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HBB ON

<400>  32
ccacgttcac cttgccccac agg                                               23


<210>  33
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HBB OFF

<400>  33
ccacattcac cttgccccac agg                                               23


<210>  34
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX ON

<400>  34
gagtccgagc agaagaagaa ggg                                               23


<210>  35
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX OFF

<400>  35
gagttagagc agaagaagaa agg                                               23


<210>  36
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HEK ON

<400>  36
ggcactgcgg ctggaggtgg ggg                                               23


<210>  37
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HEK OFF

<400>  37
tgcactgcgg ccggaggagg tgg                                               23


<210>  38
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RUNX ON

<400>  38
gcattttcag gaggaagcga tgg                                               23


<210>  39
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RUNX OFF

<400>  39
gcattttcag aaggaagcaa ggg                                               23


<210>  40
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_ON_fw

<400>  40
tggctactgg ccttatctca cagg                                              24


<210>  41
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_ON_re

<400>  41
ctctctagtc tgtgctagct cttccag                                           27


<210>  42
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_OFF1_fw

<400>  42
actcttctac cttgcacgcc tttgc                                             25


<210>  43
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_OFF1_re

<400>  43
cctgcctccc atgcaaacag tgtc                                              24


<210>  44
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_OFF2_fw

<400>  44
gagctggttt gcttatgtgt gcagg                                             25


<210>  45
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_OFF2_re

<400>  45
gcacttctcg ctggccactt acag                                              24


<210>  46
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_ON_seq_fw

<400>  46
tcgtcggcag cgtcagatgt gtataagaga caggggatca gtgaaacgca ccag             54


<210>  47
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_ON_seq_re

<400>  47
gtctcgtggg ctcggagatg tgtataagag acaggacaca cccccatttc ctgg             54


<210>  48
<211>  55
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_OFF1_seq_fw

<400>  48
tcgtcggcag cgtcagatgt gtataagaga caggactaag gcagagagac cgagg            55


<210>  49
<211>  58
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  AAVS1_OFF1_seq_re

<400>  49
gtctcgtggg ctcggagatg tgtataagag acagatgtgt gagccactta catagcac         58


<210>  50
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HBB_ON_fw

<400>  50
caagcgtccc atagactcac cctgaag                                           27


<210>  51
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HBB_ON_re

<400>  51
gtgccagaag agccaaggac aggtac                                            26


<210>  52
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HBB_OFF_fw

<400>  52
atgatctctg ccccatctat gcttgg                                            26


<210>  53
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HBB_OFF_re

<400>  53
ttgacccact gcatcagaat catttgg                                           27


<210>  54
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX_ON_fw

<400>  54
tcgtcggcag cgtcagatgt gtataagaga cagaggtgaa ggtgtggttc cag              53


<210>  55
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX_ON_re

<400>  55
gtctcgtggg ctcggagatg tgtataagag acagagtggc cagagtccag ctt              53


<210>  56
<211>  59
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX_OFF_fw

<400>  56
tcgtcggcag cgtcagatgt gtataagaga caggacacct tttaagatct gacagagaa        59


<210>  57
<211>  58
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EMX_OFF_re

<400>  57
gtctcgtggg ctcggagatg tgtataagag acagtgcaca tgtatgtaca ggagtcat         58


<210>  58
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HEK_ON_fw

<400>  58
tcgtcggcag cgtcagatgt gtataagaga caggccctcc cctcccttca aga              53


<210>  59
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HEK_ON_re

<400>  59
gtctcgtggg ctcggagatg tgtataagag acagccagac tccttctggg gcct             54


<210>  60
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HEK_OFF_fw

<400>  60
tcgtcggcag cgtcagatgt gtataagaga cagcctgggg catggcttct gag              53


<210>  61
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HEK_OFF_re

<400>  61
gtctcgtggg ctcggagatg tgtataagag acagctcaac ccaggtgttg gccc             54


<210>  62
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RUNX_ON_fw

<400>  62
tcgtcggcag cgtcagatgt gtataagaga cagtacaggc aaagctgagc aaa              53


<210>  63
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RUNX_ON_re

<400>  63
gtctcgtggg ctcggagatg tgtataagag acagccagag gtatccagca gagg             54


<210>  64
<211>  53
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RUNX_OFF_fw

<400>  64
tcgtcggcag cgtcagatgt gtataagaga caggcatgat actttggggg aga              53


<210>  65
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  RUNX_OFF_re

<400>  65
gtctcgtggg ctcggagatg tgtataagag acagtctgat cagcaatgtt gagatg           56


