                         SEQUENCE LISTING

<110>  INTELLIA THERAPEUTICS, INC.
 
<120>  POLYNUCLEOTIDES, COMPOSITIONS, AND METHODS FOR GENOME EDITING

<130>  01155-0020-00PCT

<150>  US 62/566,144
<151>  2017-09-29

<160>  198   

<170>  PatentIn version 3.5

<210>  1
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 DNA coding sequence 2

<400>  1
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcccga agaagaagag aaaggtctag     4140


<210>  2
<211>  4143
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 DNA coding sequence 1

<400>  2
atggataaga agtactcaat cgggctggat atcggaacta attccgtggg ttgggcagtg       60

atcacggatg aatacaaagt gccgtccaag aagttcaagg tcctggggaa caccgataga      120

cacagcatca agaaaaatct catcggagcc ctgctgtttg actccggcga aaccgcagaa      180

gcgacccggc tcaaacgtac cgcgaggcga cgctacaccc ggcggaagaa tcgcatctgc      240

tatctgcaag agatcttttc gaacgaaatg gcaaaggtcg acgacagctt cttccaccgc      300

ctggaagaat ctttcctggt ggaggaggac aagaagcatg aacggcatcc tatctttgga      360

aacatcgtcg acgaagtggc gtaccacgaa aagtacccga ccatctacca tctgcggaag      420

aagttggttg actcaactga caaggccgac ctcagattga tctacttggc cctcgcccat      480

atgatcaaat tccgcggaca cttcctgatc gaaggcgatc tgaaccctga taactccgac      540

gtggataagc ttttcattca actggtgcag acctacaacc aactgttcga agaaaaccca      600

atcaatgcta gcggcgtcga tgccaaggcc atcctgtccg cccggctgtc gaagtcgcgg      660

cgcctcgaaa acctgatcgc acagctgccg ggagagaaaa agaacggact tttcggcaac      720

ttgatcgctc tctcactggg actcactccc aatttcaagt ccaattttga cctggccgag      780

gacgcgaagc tgcaactctc aaaggacacc tacgacgacg acttggacaa tttgctggca      840

caaattggcg atcagtacgc ggatctgttc cttgccgcta agaacctttc ggacgcaatc      900

ttgctgtccg atatcctgcg cgtgaacacc gaaataacca aagcgccgct tagcgcctcg      960

atgattaagc ggtacgacga gcatcaccag gatctcacgc tgctcaaagc gctcgtgaga     1020

cagcaactgc ctgaaaagta caaggagatc ttcttcgacc agtccaagaa tgggtacgca     1080

gggtacatcg atggaggcgc tagccaggaa gagttctata agttcatcaa gccaatcctg     1140

gaaaagatgg acggaaccga agaactgctg gtcaagctga acagggagga tctgctccgg     1200

aaacagagaa cctttgacaa cggatccatt ccccaccaga tccatctggg tgagctgcac     1260

gccatcttgc ggcgccagga ggacttttac ccattcctca aggacaaccg ggaaaagatc     1320

gagaaaattc tgacgttccg catcccgtat tacgtgggcc cactggcgcg cggcaattcg     1380

cgcttcgcgt ggatgactag aaaatcagag gaaaccatca ctccttggaa tttcgaggaa     1440

gttgtggata agggagcttc ggcacaaagc ttcatcgaac gaatgaccaa cttcgacaag     1500

aatctcccaa acgagaaggt gcttcctaag cacagcctcc tttacgaata cttcactgtc     1560

tacaacgaac tgactaaagt gaaatacgtt actgaaggaa tgaggaagcc ggcctttctg     1620

tccggagaac agaagaaagc aattgtcgat ctgctgttca agaccaaccg caaggtgacc     1680

gtcaagcagc ttaaagagga ctacttcaag aagatcgagt gtttcgactc agtggaaatc     1740

agcggggtgg aggacagatt caacgcttcg ctgggaacct atcatgatct cctgaagatc     1800

atcaaggaca aggacttcct tgacaacgag gagaacgagg acatcctgga agatatcgtc     1860

ctgaccttga cccttttcga ggatcgcgag atgatcgagg agaggcttaa gacctacgct     1920

catctcttcg acgataaggt catgaaacaa ctcaagcgcc gccggtacac tggttggggc     1980

cgcctctccc gcaagctgat caacggtatt cgcgataaac agagcggtaa aactatcctg     2040

gatttcctca aatcggatgg cttcgctaat cgtaacttca tgcaattgat ccacgacgac     2100

agcctgacct ttaaggagga catccaaaaa gcacaagtgt ccggacaggg agactcactc     2160

catgaacaca tcgcgaatct ggccggttcg ccggcgatta agaagggaat tctgcaaact     2220

gtgaaggtgg tcgacgagct ggtgaaggtc atgggacggc acaaaccgga gaatatcgtg     2280

attgaaatgg cccgagaaaa ccagactacc cagaagggcc agaaaaactc ccgcgaaagg     2340

atgaagcgga tcgaagaagg aatcaaggag ctgggcagcc agatcctgaa agagcacccg     2400

gtggaaaaca cgcagctgca gaacgagaag ctctacctgt actatttgca aaatggacgg     2460

gacatgtacg tggaccaaga gctggacatc aatcggttgt ctgattacga cgtggaccac     2520

atcgttccac agtcctttct gaaggatgac tcgatcgata acaaggtgtt gactcgcagc     2580

gacaagaaca gagggaagtc agataatgtg ccatcggagg aggtcgtgaa gaagatgaag     2640

aattactggc ggcagctcct gaatgcgaag ctgattaccc agagaaagtt tgacaatctc     2700

actaaagccg agcgcggcgg actctcagag ctggataagg ctggattcat caaacggcag     2760

ctggtcgaga ctcggcagat taccaagcac gtggcgcaga tcttggactc ccgcatgaac     2820

actaaatacg acgagaacga taagctcatc cgggaagtga aggtgattac cctgaaaagc     2880

aaacttgtgt cggactttcg gaaggacttt cagttttaca aagtgagaga aatcaacaac     2940

taccatcacg cgcatgacgc atacctcaac gctgtggtcg gtaccgccct gatcaaaaag     3000

taccctaaac ttgaatcgga gtttgtgtac ggagactaca aggtctacga cgtgaggaag     3060

atgatagcca agtccgaaca ggaaatcggg aaagcaactg cgaaatactt cttttactca     3120

aacatcatga actttttcaa gactgaaatt acgctggcca atggagaaat caggaagagg     3180

ccactgatcg aaactaacgg agaaacgggc gaaatcgtgt gggacaaggg cagggacttc     3240

gcaactgttc gcaaagtgct ctctatgccg caagtcaata ttgtgaagaa aaccgaagtg     3300

caaaccggcg gattttcaaa ggaatcgatc ctcccaaaga gaaatagcga caagctcatt     3360

gcacgcaaga aagactggga cccgaagaag tacggaggat tcgattcgcc gactgtcgca     3420

tactccgtcc tcgtggtggc caaggtggag aagggaaaga gcaaaaagct caaatccgtc     3480

aaagagctgc tggggattac catcatggaa cgatcctcgt tcgagaagaa cccgattgat     3540

ttcctcgagg cgaagggtta caaggaggtg aagaaggatc tgatcatcaa actccccaag     3600

tactcactgt tcgaactgga aaatggtcgg aagcgcatgc tggcttcggc cggagaactc     3660

caaaaaggaa atgagctggc cttgcctagc aagtacgtca acttcctcta tcttgcttcg     3720

cactacgaaa aactcaaagg gtcaccggaa gataacgaac agaagcagct tttcgtggag     3780

cagcacaagc attatctgga tgaaatcatc gaacaaatct ccgagttttc aaagcgcgtg     3840

atcctcgccg acgccaacct cgacaaagtc ctgtcggcct acaataagca tagagataag     3900

ccgatcagag aacaggccga gaacattatc cacttgttca ccctgactaa cctgggagcc     3960

ccagccgcct tcaagtactt cgatactact atcgatcgca aaagatacac gtccaccaag     4020

gaagttctgg acgcgaccct gatccaccaa agcatcactg gactctacga aactaggatc     4080

gatctgtcgc agctgggtgg cgatggcggt ggatctccga aaaagaagag aaaggtgtaa     4140

tga                                                                   4143


<210>  3
<211>  1379
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 amino acid sequence

<400>  3

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


Gly Gly  Gly Ser Pro Lys Lys  Lys Arg Lys Val 
    1370                 1375                 


<210>  4
<211>  4140
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA open reading frame (ORF) 2

<400>  4
auggacaaga aguacagcau cggacuggac aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggagga ggaagcccga agaagaagag aaaggucuag     4140


<210>  5
<211>  4143
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA ORF 1

<400>  5
auggauaaga aguacucaau cgggcuggau aucggaacua auuccguggg uugggcagug       60

aucacggaug aauacaaagu gccguccaag aaguucaagg uccuggggaa caccgauaga      120

cacagcauca agaaaaaucu caucggagcc cugcuguuug acuccggcga aaccgcagaa      180

gcgacccggc ucaaacguac cgcgaggcga cgcuacaccc ggcggaagaa ucgcaucugc      240

uaucugcaag agaucuuuuc gaacgaaaug gcaaaggucg acgacagcuu cuuccaccgc      300

cuggaagaau cuuuccuggu ggaggaggac aagaagcaug aacggcaucc uaucuuugga      360

aacaucgucg acgaaguggc guaccacgaa aaguacccga ccaucuacca ucugcggaag      420

aaguugguug acucaacuga caaggccgac cucagauuga ucuacuuggc ccucgcccau      480

augaucaaau uccgcggaca cuuccugauc gaaggcgauc ugaacccuga uaacuccgac      540

guggauaagc uuuucauuca acuggugcag accuacaacc aacuguucga agaaaaccca      600

aucaaugcua gcggcgucga ugccaaggcc auccuguccg cccggcuguc gaagucgcgg      660

cgccucgaaa accugaucgc acagcugccg ggagagaaaa agaacggacu uuucggcaac      720

uugaucgcuc ucucacuggg acucacuccc aauuucaagu ccaauuuuga ccuggccgag      780

gacgcgaagc ugcaacucuc aaaggacacc uacgacgacg acuuggacaa uuugcuggca      840

caaauuggcg aucaguacgc ggaucuguuc cuugccgcua agaaccuuuc ggacgcaauc      900

uugcuguccg auauccugcg cgugaacacc gaaauaacca aagcgccgcu uagcgccucg      960

augauuaagc gguacgacga gcaucaccag gaucucacgc ugcucaaagc gcucgugaga     1020

cagcaacugc cugaaaagua caaggagauc uucuucgacc aguccaagaa uggguacgca     1080

ggguacaucg auggaggcgc uagccaggaa gaguucuaua aguucaucaa gccaauccug     1140

gaaaagaugg acggaaccga agaacugcug gucaagcuga acagggagga ucugcuccgg     1200

aaacagagaa ccuuugacaa cggauccauu ccccaccaga uccaucuggg ugagcugcac     1260

gccaucuugc ggcgccagga ggacuuuuac ccauuccuca aggacaaccg ggaaaagauc     1320

gagaaaauuc ugacguuccg caucccguau uacgugggcc cacuggcgcg cggcaauucg     1380

cgcuucgcgu ggaugacuag aaaaucagag gaaaccauca cuccuuggaa uuucgaggaa     1440

guuguggaua agggagcuuc ggcacaaagc uucaucgaac gaaugaccaa cuucgacaag     1500

aaucucccaa acgagaaggu gcuuccuaag cacagccucc uuuacgaaua cuucacuguc     1560

uacaacgaac ugacuaaagu gaaauacguu acugaaggaa ugaggaagcc ggccuuucug     1620

uccggagaac agaagaaagc aauugucgau cugcuguuca agaccaaccg caaggugacc     1680

gucaagcagc uuaaagagga cuacuucaag aagaucgagu guuucgacuc aguggaaauc     1740

agcggggugg aggacagauu caacgcuucg cugggaaccu aucaugaucu ccugaagauc     1800

aucaaggaca aggacuuccu ugacaacgag gagaacgagg acauccugga agauaucguc     1860

cugaccuuga cccuuuucga ggaucgcgag augaucgagg agaggcuuaa gaccuacgcu     1920

caucucuucg acgauaaggu caugaaacaa cucaagcgcc gccgguacac ugguuggggc     1980

cgccucuccc gcaagcugau caacgguauu cgcgauaaac agagcgguaa aacuauccug     2040

gauuuccuca aaucggaugg cuucgcuaau cguaacuuca ugcaauugau ccacgacgac     2100

agccugaccu uuaaggagga cauccaaaaa gcacaagugu ccggacaggg agacucacuc     2160

caugaacaca ucgcgaaucu ggccgguucg ccggcgauua agaagggaau ucugcaaacu     2220

gugaaggugg ucgacgagcu ggugaagguc augggacggc acaaaccgga gaauaucgug     2280

auugaaaugg cccgagaaaa ccagacuacc cagaagggcc agaaaaacuc ccgcgaaagg     2340

augaagcgga ucgaagaagg aaucaaggag cugggcagcc agauccugaa agagcacccg     2400

guggaaaaca cgcagcugca gaacgagaag cucuaccugu acuauuugca aaauggacgg     2460

gacauguacg uggaccaaga gcuggacauc aaucgguugu cugauuacga cguggaccac     2520

aucguuccac aguccuuucu gaaggaugac ucgaucgaua acaagguguu gacucgcagc     2580

gacaagaaca gagggaaguc agauaaugug ccaucggagg aggucgugaa gaagaugaag     2640

aauuacuggc ggcagcuccu gaaugcgaag cugauuaccc agagaaaguu ugacaaucuc     2700

acuaaagccg agcgcggcgg acucucagag cuggauaagg cuggauucau caaacggcag     2760

cuggucgaga cucggcagau uaccaagcac guggcgcaga ucuuggacuc ccgcaugaac     2820

acuaaauacg acgagaacga uaagcucauc cgggaaguga aggugauuac ccugaaaagc     2880

aaacuugugu cggacuuucg gaaggacuuu caguuuuaca aagugagaga aaucaacaac     2940

uaccaucacg cgcaugacgc auaccucaac gcuguggucg guaccgcccu gaucaaaaag     3000

uacccuaaac uugaaucgga guuuguguac ggagacuaca aggucuacga cgugaggaag     3060

augauagcca aguccgaaca ggaaaucggg aaagcaacug cgaaauacuu cuuuuacuca     3120

aacaucauga acuuuuucaa gacugaaauu acgcuggcca auggagaaau caggaagagg     3180

ccacugaucg aaacuaacgg agaaacgggc gaaaucgugu gggacaaggg cagggacuuc     3240

gcaacuguuc gcaaagugcu cucuaugccg caagucaaua uugugaagaa aaccgaagug     3300

caaaccggcg gauuuucaaa ggaaucgauc cucccaaaga gaaauagcga caagcucauu     3360

gcacgcaaga aagacuggga cccgaagaag uacggaggau ucgauucgcc gacugucgca     3420

uacuccgucc ucgugguggc caagguggag aagggaaaga gcaaaaagcu caaauccguc     3480

aaagagcugc uggggauuac caucauggaa cgauccucgu ucgagaagaa cccgauugau     3540

uuccucgagg cgaaggguua caaggaggug aagaaggauc ugaucaucaa acuccccaag     3600

uacucacugu ucgaacugga aaauggucgg aagcgcaugc uggcuucggc cggagaacuc     3660

caaaaaggaa augagcuggc cuugccuagc aaguacguca acuuccucua ucuugcuucg     3720

cacuacgaaa aacucaaagg gucaccggaa gauaacgaac agaagcagcu uuucguggag     3780

cagcacaagc auuaucugga ugaaaucauc gaacaaaucu ccgaguuuuc aaagcgcgug     3840

auccucgccg acgccaaccu cgacaaaguc cugucggccu acaauaagca uagagauaag     3900

ccgaucagag aacaggccga gaacauuauc cacuuguuca cccugacuaa ccugggagcc     3960

ccagccgccu ucaaguacuu cgauacuacu aucgaucgca aaagauacac guccaccaag     4020

gaaguucugg acgcgacccu gauccaccaa agcaucacug gacucuacga aacuaggauc     4080

gaucugucgc agcugggugg cgauggcggu ggaucuccga aaaagaagag aaagguguaa     4140

uga                                                                   4143


<210>  6
<211>  1379
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase (D10A) amino acid sequence

<400>  6

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


Gly Gly  Gly Ser Pro Lys Lys  Lys Arg Lys Val 
    1370                 1375                 


<210>  7
<211>  4140
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase (D10A) mRNA ORF

<400>  7
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggagga ggaagcccga agaagaagag aaaggucuag     4140


<210>  8
<211>  1379
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 (D10A H840A) amino acid sequence

<400>  8

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


Gly Gly  Gly Ser Pro Lys Lys  Lys Arg Lys Val 
    1370                 1375                 


<210>  9
<211>  4140
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 (D10A H840A) mRNA ORF

<400>  9
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgacgca     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggagga ggaagcccga agaagaagag aaaggucuag     4140


<210>  10
<211>  4134
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 bare coding sequence

<400>  10
gacaagaagu acagcaucgg acuggacauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agcccgaaga agaagagaaa gguc           4134


<210>  11
<211>  4134
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase bare coding sequence

<400>  11
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agcccgaaga agaagagaaa gguc           4134


<210>  12
<211>  4134
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 bare coding sequence

<400>  12
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgacgcaauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agcccgaaga agaagagaaa gguc           4134


<210>  13
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 (without NLS)

<400>  13

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  14
<211>  4107
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA ORF encoding SEQ ID NO: 13 using minimal 
       uridine codons as listed in Table 3, with start and stop codons

<400>  14
auggacaaga aguacagcau cggacuggac aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacuag                                         4107


<210>  15
<211>  4101
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 coding sequence encoding SEQ ID NO: 13 using 
       minimal uridine codons as listed in Table 3 (no start or stop 
       codons; suitable for inclusion in fusion protein coding sequence)

<400>  15
gacaagaagu acagcaucgg acuggacauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga c                                               4101


<210>  16
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 nickase (without NLS)

<400>  16

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  17
<211>  4107
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase mRNA ORF encoding SEQ ID NO: 16 using 
       minimal uridine codons as listed in Table 3, with start and stop 
       codons

<400>  17
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacuag                                         4107


<210>  18
<211>  4101
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase coding sequence encoding SEQ ID NO: 16 
       using minimal uridine codons as listed in Table 3 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  18
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga c                                               4101


<210>  19
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of dCas9 (without NLS)

<400>  19

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  20
<211>  4107
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 mRNA ORF encoding SEQ ID NO: 19 using minimal 
       uridine codons as listed in Table 3, with start and stop codons

<400>  20
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgacgca     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacuag                                         4107


<210>  21
<211>  4113
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 coding sequence encoding SEQ ID NO: 19 using 
       minimal uridine codons as listed in Table 3 (no start or stop 
       codons; suitable for inclusion in fusion protein coding sequence)

<400>  21
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgacgcaauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaggagga agc                                  4113


<210>  22
<211>  1392
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 with two nuclear 
       localization signals as the C-terminal amino acids

<400>  22

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


Gly Ser  Gly Ser Pro Lys Lys  Lys Arg Lys Val Asp  Gly Ser Pro 
    1370                 1375                 1380             


Lys Lys  Lys Arg Lys Val Asp  Ser Gly 
    1385                 1390         


<210>  23
<211>  4179
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 mRNA ORF encoding SEQ ID NO: 22 using minimal 
       uridine codons as listed in Table 3, with start and stop codons

<400>  23
auggacaaga aguacagcau cggacuggac aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggaagc ggaagcccga agaagaagag aaaggucgac     4140

ggaagcccga agaagaagag aaaggucgac agcggauag                            4179


<210>  24
<211>  4173
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 coding sequence encoding SEQ ID NO: 23 using 
       minimal uridine codons as listed in Table 3 (no start or stop 
       codons; suitable for inclusion in fusion protein coding sequence)

<400>  24
gacaagaagu acagcaucgg acuggacauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaagcgga agcccgaaga agaagagaaa ggucgacgga     4140

agcccgaaga agaagagaaa ggucgacagc gga                                  4173


<210>  25
<211>  1392
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Cas9 nickase with two nuclear 
       localization signals as the C-terminal amino acids

<400>  25

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


Gly Ser  Gly Ser Pro Lys Lys  Lys Arg Lys Val Asp  Gly Ser Pro 
    1370                 1375                 1380             


Lys Lys  Lys Arg Lys Val Asp  Ser Gly 
    1385                 1390         


<210>  26
<211>  4179
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase mRNA ORF encoding SEQ ID NO: 25 using 
       minimal uridine codons as listed in Table 3, with start and stop 
       codons

<400>  26
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgaccac     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggaagc ggaagcccga agaagaagag aaaggucgac     4140

ggaagcccga agaagaagag aaaggucgac agcggauag                            4179


<210>  27
<211>  4173
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase coding sequence encoding SEQ ID NO: 25 
       using minimal uridine codons as listed in Table 3 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  27
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgaccacauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaagcgga agcccgaaga agaagagaaa ggucgacgga     4140

agcccgaaga agaagagaaa ggucgacagc gga                                  4173


<210>  28
<211>  1392
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of dCas9 with two nuclear 
       localization signals as the C-terminal amino acids

<400>  28

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


Gly Ser  Gly Ser Pro Lys Lys  Lys Arg Lys Val Asp  Gly Ser Pro 
    1370                 1375                 1380             


Lys Lys  Lys Arg Lys Val Asp  Ser Gly 
    1385                 1390         


<210>  29
<211>  4179
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 mRNA ORF encoding SEQ ID NO: 28 using minimal 
       uridine codons as listed in Table 3, with start and stop codons

<400>  29
auggacaaga aguacagcau cggacuggca aucggaacaa acagcgucgg augggcaguc       60

aucacagacg aauacaaggu cccgagcaag aaguucaagg uccugggaaa cacagacaga      120

cacagcauca agaagaaccu gaucggagca cugcuguucg acagcggaga aacagcagaa      180

gcaacaagac ugaagagaac agcaagaaga agauacacaa gaagaaagaa cagaaucugc      240

uaccugcagg aaaucuucag caacgaaaug gcaaaggucg acgacagcuu cuuccacaga      300

cuggaagaaa gcuuccuggu cgaagaagac aagaagcacg aaagacaccc gaucuucgga      360

aacaucgucg acgaagucgc auaccacgaa aaguacccga caaucuacca ccugagaaag      420

aagcuggucg acagcacaga caaggcagac cugagacuga ucuaccuggc acuggcacac      480

augaucaagu ucagaggaca cuuccugauc gaaggagacc ugaacccgga caacagcgac      540

gucgacaagc uguucaucca gcugguccag acauacaacc agcuguucga agaaaacccg      600

aucaacgcaa gcggagucga cgcaaaggca auccugagcg caagacugag caagagcaga      660

agacuggaaa accugaucgc acagcugccg ggagaaaaga agaacggacu guucggaaac      720

cugaucgcac ugagccuggg acugacaccg aacuucaaga gcaacuucga ccuggcagaa      780

gacgcaaagc ugcagcugag caaggacaca uacgacgacg accuggacaa ccugcuggca      840

cagaucggag accaguacgc agaccuguuc cuggcagcaa agaaccugag cgacgcaauc      900

cugcugagcg acauccugag agucaacaca gaaaucacaa aggcaccgcu gagcgcaagc      960

augaucaaga gauacgacga acaccaccag gaccugacac ugcugaaggc acuggucaga     1020

cagcagcugc cggaaaagua caaggaaauc uucuucgacc agagcaagaa cggauacgca     1080

ggauacaucg acggaggagc aagccaggaa gaauucuaca aguucaucaa gccgauccug     1140

gaaaagaugg acggaacaga agaacugcug gucaagcuga acagagaaga ccugcugaga     1200

aagcagagaa cauucgacaa cggaagcauc ccgcaccaga uccaccuggg agaacugcac     1260

gcaauccuga gaagacagga agacuucuac ccguuccuga aggacaacag agaaaagauc     1320

gaaaagaucc ugacauucag aaucccguac uacgucggac cgcuggcaag aggaaacagc     1380

agauucgcau ggaugacaag aaagagcgaa gaaacaauca caccguggaa cuucgaagaa     1440

gucgucgaca agggagcaag cgcacagagc uucaucgaaa gaaugacaaa cuucgacaag     1500

aaccugccga acgaaaaggu ccugccgaag cacagccugc uguacgaaua cuucacaguc     1560

uacaacgaac ugacaaaggu caaguacguc acagaaggaa ugagaaagcc ggcauuccug     1620

agcggagaac agaagaaggc aaucgucgac cugcuguuca agacaaacag aaaggucaca     1680

gucaagcagc ugaaggaaga cuacuucaag aagaucgaau gcuucgacag cgucgaaauc     1740

agcggagucg aagacagauu caacgcaagc cugggaacau accacgaccu gcugaagauc     1800

aucaaggaca aggacuuccu ggacaacgaa gaaaacgaag acauccugga agacaucguc     1860

cugacacuga cacuguucga agacagagaa augaucgaag aaagacugaa gacauacgca     1920

caccuguucg acgacaaggu caugaagcag cugaagagaa gaagauacac aggaugggga     1980

agacugagca gaaagcugau caacggaauc agagacaagc agagcggaaa gacaauccug     2040

gacuuccuga agagcgacgg auucgcaaac agaaacuuca ugcagcugau ccacgacgac     2100

agccugacau ucaaggaaga cauccagaag gcacagguca gcggacaggg agacagccug     2160

cacgaacaca ucgcaaaccu ggcaggaagc ccggcaauca agaagggaau ccugcagaca     2220

gucaaggucg ucgacgaacu ggucaagguc augggaagac acaagccgga aaacaucguc     2280

aucgaaaugg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

augaagagaa ucgaagaagg aaucaaggaa cugggaagcc agauccugaa ggaacacccg     2400

gucgaaaaca cacagcugca gaacgaaaag cuguaccugu acuaccugca gaacggaaga     2460

gacauguacg ucgaccagga acuggacauc aacagacuga gcgacuacga cgucgacgca     2520

aucgucccgc agagcuuccu gaaggacgac agcaucgaca acaagguccu gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacguc ccgagcgaag aagucgucaa gaagaugaag     2640

aacuacugga gacagcugcu gaacgcaaag cugaucacac agagaaaguu cgacaaccug     2700

acaaaggcag agagaggagg acugagcgaa cuggacaagg caggauucau caagagacag     2760

cuggucgaaa caagacagau cacaaagcac gucgcacaga uccuggacag cagaaugaac     2820

acaaaguacg acgaaaacga caagcugauc agagaaguca aggucaucac acugaagagc     2880

aagcugguca gcgacuucag aaaggacuuc caguucuaca aggucagaga aaucaacaac     2940

uaccaccacg cacacgacgc auaccugaac gcagucgucg gaacagcacu gaucaagaag     3000

uacccgaagc uggaaagcga auucgucuac ggagacuaca aggucuacga cgucagaaag     3060

augaucgcaa agagcgaaca ggaaaucgga aaggcaacag caaaguacuu cuucuacagc     3120

aacaucauga acuucuucaa gacagaaauc acacuggcaa acggagaaau cagaaagaga     3180

ccgcugaucg aaacaaacgg agaaacagga gaaaucgucu gggacaaggg aagagacuuc     3240

gcaacaguca gaaagguccu gagcaugccg caggucaaca ucgucaagaa gacagaaguc     3300

cagacaggag gauucagcaa ggaaagcauc cugccgaaga gaaacagcga caagcugauc     3360

gcaagaaaga aggacuggga cccgaagaag uacggaggau ucgacagccc gacagucgca     3420

uacagcgucc uggucgucgc aaaggucgaa aagggaaaga gcaagaagcu gaagagcguc     3480

aaggaacugc ugggaaucac aaucauggaa agaagcagcu ucgaaaagaa cccgaucgac     3540

uuccuggaag caaagggaua caaggaaguc aagaaggacc ugaucaucaa gcugccgaag     3600

uacagccugu ucgaacugga aaacggaaga aagagaaugc uggcaagcgc aggagaacug     3660

cagaagggaa acgaacuggc acugccgagc aaguacguca acuuccugua ccuggcaagc     3720

cacuacgaaa agcugaaggg aagcccggaa gacaacgaac agaagcagcu guucgucgaa     3780

cagcacaagc acuaccugga cgaaaucauc gaacagauca gcgaauucag caagagaguc     3840

auccuggcag acgcaaaccu ggacaagguc cugagcgcau acaacaagca cagagacaag     3900

ccgaucagag aacaggcaga aaacaucauc caccuguuca cacugacaaa ccugggagca     3960

ccggcagcau ucaaguacuu cgacacaaca aucgacagaa agagauacac aagcacaaag     4020

gaaguccugg acgcaacacu gauccaccag agcaucacag gacuguacga aacaagaauc     4080

gaccugagcc agcugggagg agacggaagc ggaagcccga agaagaagag aaaggucgac     4140

ggaagcccga agaagaagag aaaggucgac agcggauag                            4179


<210>  30
<211>  4173
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 coding sequence encoding SEQ ID NO: 28 using 
       minimal uridine codons as listed in Table 3 (no start or stop 
       codons; suitable for inclusion in fusion protein coding sequence)

<400>  30
gacaagaagu acagcaucgg acuggcaauc ggaacaaaca gcgucggaug ggcagucauc       60

acagacgaau acaagguccc gagcaagaag uucaaggucc ugggaaacac agacagacac      120

agcaucaaga agaaccugau cggagcacug cuguucgaca gcggagaaac agcagaagca      180

acaagacuga agagaacagc aagaagaaga uacacaagaa gaaagaacag aaucugcuac      240

cugcaggaaa ucuucagcaa cgaaauggca aaggucgacg acagcuucuu ccacagacug      300

gaagaaagcu uccuggucga agaagacaag aagcacgaaa gacacccgau cuucggaaac      360

aucgucgacg aagucgcaua ccacgaaaag uacccgacaa ucuaccaccu gagaaagaag      420

cuggucgaca gcacagacaa ggcagaccug agacugaucu accuggcacu ggcacacaug      480

aucaaguuca gaggacacuu ccugaucgaa ggagaccuga acccggacaa cagcgacguc      540

gacaagcugu ucauccagcu gguccagaca uacaaccagc uguucgaaga aaacccgauc      600

aacgcaagcg gagucgacgc aaaggcaauc cugagcgcaa gacugagcaa gagcagaaga      660

cuggaaaacc ugaucgcaca gcugccggga gaaaagaaga acggacuguu cggaaaccug      720

aucgcacuga gccugggacu gacaccgaac uucaagagca acuucgaccu ggcagaagac      780

gcaaagcugc agcugagcaa ggacacauac gacgacgacc uggacaaccu gcuggcacag      840

aucggagacc aguacgcaga ccuguuccug gcagcaaaga accugagcga cgcaauccug      900

cugagcgaca uccugagagu caacacagaa aucacaaagg caccgcugag cgcaagcaug      960

aucaagagau acgacgaaca ccaccaggac cugacacugc ugaaggcacu ggucagacag     1020

cagcugccgg aaaaguacaa ggaaaucuuc uucgaccaga gcaagaacgg auacgcagga     1080

uacaucgacg gaggagcaag ccaggaagaa uucuacaagu ucaucaagcc gauccuggaa     1140

aagauggacg gaacagaaga acugcugguc aagcugaaca gagaagaccu gcugagaaag     1200

cagagaacau ucgacaacgg aagcaucccg caccagaucc accugggaga acugcacgca     1260

auccugagaa gacaggaaga cuucuacccg uuccugaagg acaacagaga aaagaucgaa     1320

aagauccuga cauucagaau cccguacuac gucggaccgc uggcaagagg aaacagcaga     1380

uucgcaugga ugacaagaaa gagcgaagaa acaaucacac cguggaacuu cgaagaaguc     1440

gucgacaagg gagcaagcgc acagagcuuc aucgaaagaa ugacaaacuu cgacaagaac     1500

cugccgaacg aaaagguccu gccgaagcac agccugcugu acgaauacuu cacagucuac     1560

aacgaacuga caaaggucaa guacgucaca gaaggaauga gaaagccggc auuccugagc     1620

ggagaacaga agaaggcaau cgucgaccug cuguucaaga caaacagaaa ggucacaguc     1680

aagcagcuga aggaagacua cuucaagaag aucgaaugcu ucgacagcgu cgaaaucagc     1740

ggagucgaag acagauucaa cgcaagccug ggaacauacc acgaccugcu gaagaucauc     1800

aaggacaagg acuuccugga caacgaagaa aacgaagaca uccuggaaga caucguccug     1860

acacugacac uguucgaaga cagagaaaug aucgaagaaa gacugaagac auacgcacac     1920

cuguucgacg acaaggucau gaagcagcug aagagaagaa gauacacagg auggggaaga     1980

cugagcagaa agcugaucaa cggaaucaga gacaagcaga gcggaaagac aauccuggac     2040

uuccugaaga gcgacggauu cgcaaacaga aacuucaugc agcugaucca cgacgacagc     2100

cugacauuca aggaagacau ccagaaggca caggucagcg gacagggaga cagccugcac     2160

gaacacaucg caaaccuggc aggaagcccg gcaaucaaga agggaauccu gcagacaguc     2220

aaggucgucg acgaacuggu caaggucaug ggaagacaca agccggaaaa caucgucauc     2280

gaaauggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaaug     2340

aagagaaucg aagaaggaau caaggaacug ggaagccaga uccugaagga acacccgguc     2400

gaaaacacac agcugcagaa cgaaaagcug uaccuguacu accugcagaa cggaagagac     2460

auguacgucg accaggaacu ggacaucaac agacugagcg acuacgacgu cgacgcaauc     2520

gucccgcaga gcuuccugaa ggacgacagc aucgacaaca agguccugac aagaagcgac     2580

aagaacagag gaaagagcga caacgucccg agcgaagaag ucgucaagaa gaugaagaac     2640

uacuggagac agcugcugaa cgcaaagcug aucacacaga gaaaguucga caaccugaca     2700

aaggcagaga gaggaggacu gagcgaacug gacaaggcag gauucaucaa gagacagcug     2760

gucgaaacaa gacagaucac aaagcacguc gcacagaucc uggacagcag aaugaacaca     2820

aaguacgacg aaaacgacaa gcugaucaga gaagucaagg ucaucacacu gaagagcaag     2880

cuggucagcg acuucagaaa ggacuuccag uucuacaagg ucagagaaau caacaacuac     2940

caccacgcac acgacgcaua ccugaacgca gucgucggaa cagcacugau caagaaguac     3000

ccgaagcugg aaagcgaauu cgucuacgga gacuacaagg ucuacgacgu cagaaagaug     3060

aucgcaaaga gcgaacagga aaucggaaag gcaacagcaa aguacuucuu cuacagcaac     3120

aucaugaacu ucuucaagac agaaaucaca cuggcaaacg gagaaaucag aaagagaccg     3180

cugaucgaaa caaacggaga aacaggagaa aucgucuggg acaagggaag agacuucgca     3240

acagucagaa agguccugag caugccgcag gucaacaucg ucaagaagac agaaguccag     3300

acaggaggau ucagcaagga aagcauccug ccgaagagaa acagcgacaa gcugaucgca     3360

agaaagaagg acugggaccc gaagaaguac ggaggauucg acagcccgac agucgcauac     3420

agcguccugg ucgucgcaaa ggucgaaaag ggaaagagca agaagcugaa gagcgucaag     3480

gaacugcugg gaaucacaau cauggaaaga agcagcuucg aaaagaaccc gaucgacuuc     3540

cuggaagcaa agggauacaa ggaagucaag aaggaccuga ucaucaagcu gccgaaguac     3600

agccuguucg aacuggaaaa cggaagaaag agaaugcugg caagcgcagg agaacugcag     3660

aagggaaacg aacuggcacu gccgagcaag uacgucaacu uccuguaccu ggcaagccac     3720

uacgaaaagc ugaagggaag cccggaagac aacgaacaga agcagcuguu cgucgaacag     3780

cacaagcacu accuggacga aaucaucgaa cagaucagcg aauucagcaa gagagucauc     3840

cuggcagacg caaaccugga caagguccug agcgcauaca acaagcacag agacaagccg     3900

aucagagaac aggcagaaaa caucauccac cuguucacac ugacaaaccu gggagcaccg     3960

gcagcauuca aguacuucga cacaacaauc gacagaaaga gauacacaag cacaaaggaa     4020

guccuggacg caacacugau ccaccagagc aucacaggac uguacgaaac aagaaucgac     4080

cugagccagc ugggaggaga cggaagcgga agcccgaaga agaagagaaa ggucgacgga     4140

agcccgaaga agaagagaaa ggucgacagc gga                                  4173


<210>  31
<211>  17
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: T7 promoter

<400>  31
taatacgact cactata                                                      17


<210>  32
<211>  50
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(50)
<223>  Human beta-globin 5 UTR

<400>  32
acatttgctt ctgacacaac tgtgttcact agcaacctca aacagacacc                  50


<210>  33
<211>  132
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(132)
<223>  Human beta-globin 3 UTR

<400>  33
gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa gtccaactac       60

taaactgggg gatattatga agggccttga gcatctggat tctgcctaat aaaaaacatt      120

tattttcatt gc                                                          132


<210>  34
<211>  66
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(66)
<223>  Human alpha-globin 5 UTR

<400>  34
cataaaccct ggcgcgctcg cggcccggca ctcttctggt ccccacagac tcagagagaa       60

cccacc                                                                  66


<210>  35
<211>  110
<212>  DNA
<213>  Homo sapiens


<220>
<221>  misc_feature
<222>  (1)..(110)
<223>  Human alpha-globin 3 UTR

<400>  35
gctggagcct cggtggccat gcttcttgcc ccttgggcct ccccccagcc cctcctcccc       60

ttcctgcacc cgtacccccg tggtctttga ataaagtctg agtgggcggc                 110


<210>  36
<211>  29
<212>  DNA
<213>  Xenopus laevis


<220>
<221>  misc_feature
<222>  (1)..(29)
<223>  Xenopus laevis beta-globin 5 UTR

<400>  36
aagctcagaa taaacgctca actttggcc                                         29


<210>  37
<211>  130
<212>  DNA
<213>  Xenopus laevis


<220>
<221>  misc_feature
<222>  (1)..(130)
<223>  Xenopus laevis beta-globin 3 UTR

<400>  37
accagcctca agaacacccg aatggagtct ctaagctaca taataccaac ttacacttta       60

caaaatgttg tcccccaaaa tgtagccatt cgtatctgct cctaataaaa agaaagtttc      120

ttcacattct                                                             130


<210>  38
<211>  27
<212>  DNA
<213>  Bos taurus


<220>
<221>  misc_feature
<222>  (1)..(27)
<223>  Bovine Growth Hormone 5 UTR

<400>  38
cagggtcctg tggacagctc accagct                                           27


<210>  39
<211>  102
<212>  DNA
<213>  Bos taurus


<220>
<221>  misc_feature
<222>  (1)..(102)
<223>  Bovine Growth Hormone 3 UTR

<400>  39
ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac       60

tcccactgtc ctttcctaat aaaatgagga aattgcatcg ca                         102


<210>  40
<211>  93
<212>  DNA
<213>  Mus musculus


<220>
<221>  misc_feature
<222>  (1)..(93)
<223>  Mus musculus hemoglobin alpha, adult chain 1 (Hba-a1), 3UTR

<400>  40
gctgccttct gcggggcttg ccttctggcc atgcccttct tctctccctt gcacctgtac       60

ctcttggtct ttgaataaag cctgagtagg aag                                    93


<210>  41
<211>  61
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: HSD17B4 5 UTR

<400>  41
tcccgcagtc ggcgtccagc ggctctgctt gttcgtgtgt gtgtcgttgc aggccttatt       60

c                                                                       61


<210>  42
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G282 guide RNA targeting TTR


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  42
uuacagccac gucuacagca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  43
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 4, Kozak sequence, and 3 UTR of ALB

<400>  43
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  44
<211>  4405
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 4, and 3 UTR of ALB

<400>  44
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg      120

atgggcagtc atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa      180

cacagacaga cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga      240

aacagcagaa gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa      300

cagaatctgc tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt      360

cttccacaga ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc      420

gatcttcgga aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca      480

cctgagaaag aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc      540

actggcacac atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga      600

caacagcgac gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga      660

agaaaacccg atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag      720

caagagcaga agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact      780

gttcggaaac ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga      840

cctggcagaa gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa      900

cctgctggca cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag      960

cgacgcaatc ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct     1020

gagcgcaagc atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc     1080

actggtcaga cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa     1140

cggatacgca ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa     1200

gccgatcctg gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga     1260

cctgctgaga aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg     1320

agaactgcac gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag     1380

agaaaagatc gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag     1440

aggaaacagc agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa     1500

cttcgaagaa gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa     1560

cttcgacaag aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata     1620

cttcacagtc tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc     1680

ggcattcctg agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag     1740

aaaggtcaca gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag     1800

cgtcgaaatc agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct     1860

gctgaagatc atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga     1920

agacatcgtc ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa     1980

gacatacgca cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac     2040

aggatgggga agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa     2100

gacaatcctg gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat     2160

ccacgacgac agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg     2220

agacagcctg cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat     2280

cctgcagaca gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga     2340

aaacatcgtc atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag     2400

cagagaaaga atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa     2460

ggaacacccg gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca     2520

gaacggaaga gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga     2580

cgtcgaccac atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct     2640

gacaagaagc gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa     2700

gaagatgaag aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt     2760

cgacaacctg acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat     2820

caagagacag ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag     2880

cagaatgaac acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac     2940

actgaagagc aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga     3000

aatcaacaac taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact     3060

gatcaagaag tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga     3120

cgtcagaaag atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt     3180

cttctacagc aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat     3240

cagaaagaga ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg     3300

aagagacttc gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa     3360

gacagaagtc cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga     3420

caagctgatc gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc     3480

gacagtcgca tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct     3540

gaagagcgtc aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa     3600

cccgatcgac ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa     3660

gctgccgaag tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc     3720

aggagaactg cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta     3780

cctggcaagc cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct     3840

gttcgtcgaa cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag     3900

caagagagtc atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca     3960

cagagacaag ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa     4020

cctgggagca ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac     4080

aagcacaaag gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga     4140

aacaagaatc gacctgagcc agctgggagg agacggagga ggaagcccga agaagaagag     4200

aaaggtctag ctagccatca catttaaaag catctcagcc taccatgaga ataagagaaa     4260

gaaaatgaag atcaatagct tattcatctc tttttctttt tcgttggtgt aaagccaaca     4320

ccctgtctaa aaaacataaa tttctttaat cattttgcct cttttctctg tgcttcaatt     4380

aataaaaaat ggaaagaacc tcgag                                           4405


<210>  45
<211>  4188
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Alternative Cas9 ORF with 19.36% U content

<400>  45
atggataaga agtactcgat cgggctggat atcggaacta attccgtggg ttgggcagtg       60

atcacggatg aatacaaagt gccgtccaag aagttcaagg tcctggggaa caccgataga      120

cacagcatca agaagaatct catcggagcc ctgctgtttg actccggcga aaccgcagaa      180

gcgacccggc tcaaacgtac cgcgaggcga cgctacaccc ggcggaagaa tcgcatctgc      240

tatctgcaag aaatcttttc gaacgaaatg gcaaaggtgg acgacagctt cttccaccgc      300

ctggaagaat ctttcctggt ggaggaggac aagaagcatg aacggcatcc tatctttgga      360

aacatcgtgg acgaagtggc gtaccacgaa aagtacccga ccatctacca tctgcggaag      420

aagttggttg actcaactga caaggccgac ctcagattga tctacttggc cctcgcccat      480

atgatcaaat tccgcggaca cttcctgatc gaaggcgatc tgaaccctga taactccgac      540

gtggataagc tgttcattca actggtgcag acctacaacc aactgttcga agaaaaccca      600

atcaatgcca gcggcgtcga tgccaaggcc atcctgtccg cccggctgtc gaagtcgcgg      660

cgcctcgaaa acctgatcgc acagctgccg ggagagaaga agaacggact tttcggcaac      720

ttgatcgctc tctcactggg actcactccc aatttcaagt ccaattttga cctggccgag      780

gacgcgaagc tgcaactctc aaaggacacc tacgacgacg acttggacaa tttgctggca      840

caaattggcg atcagtacgc ggatctgttc cttgccgcta agaacctttc ggacgcaatc      900

ttgctgtccg atatcctgcg cgtgaacacc gaaataacca aagcgccgct tagcgcctcg      960

atgattaagc ggtacgacga gcatcaccag gatctcacgc tgctcaaagc gctcgtgaga     1020

cagcaactgc ctgaaaagta caaggagatt ttcttcgacc agtccaagaa tgggtacgca     1080

gggtacatcg atggaggcgc cagccaggaa gagttctata agttcatcaa gccaatcctg     1140

gaaaagatgg acggaaccga agaactgctg gtcaagctga acagggagga tctgctccgc     1200

aaacagagaa cctttgacaa cggaagcatt ccacaccaga tccatctggg tgagctgcac     1260

gccatcttgc ggcgccagga ggacttttac ccattcctca aggacaaccg ggaaaagatc     1320

gagaaaattc tgacgttccg catcccgtat tacgtgggcc cactggcgcg cggcaattcg     1380

cgcttcgcgt ggatgactag aaaatcagag gaaaccatca ctccttggaa tttcgaggaa     1440

gttgtggata agggagcttc ggcacaatcc ttcatcgaac gaatgaccaa cttcgacaag     1500

aatctcccaa acgagaaggt gcttcctaag cacagcctcc tttacgaata cttcactgtc     1560

tacaacgaac tgactaaagt gaaatacgtt actgaaggaa tgaggaagcc ggcctttctg     1620

agcggagaac agaagaaagc gattgtcgat ctgctgttca agaccaaccg caaggtgacc     1680

gtcaagcagc ttaaagagga ctacttcaag aagatcgagt gtttcgactc agtggaaatc     1740

agcggagtgg aggacagatt caacgcttcg ctgggaacct atcatgatct cctgaagatc     1800

atcaaggaca aggacttcct tgacaacgag gagaacgagg acatcctgga agatatcgtc     1860

ctgaccttga cccttttcga ggatcgcgag atgatcgagg agaggcttaa gacctacgct     1920

catctcttcg acgataaggt catgaaacaa ctcaagcgcc gccggtacac tggttggggc     1980

cgcctctccc gcaagctgat caacggtatt cgcgataaac agagcggtaa aactatcctg     2040

gatttcctca aatcggatgg cttcgctaat cgtaacttca tgcagttgat ccacgacgac     2100

agcctgacct ttaaggagga catccagaaa gcacaagtga gcggacaggg agactcactc     2160

catgaacaca tcgcgaatct ggccggttcg ccggcgatta agaagggaat cctgcaaact     2220

gtgaaggtgg tggacgagct ggtgaaggtc atgggacggc acaaaccgga gaatatcgtg     2280

attgaaatgg cccgagaaaa ccagactacc cagaagggcc agaagaactc ccgcgaaagg     2340

atgaagcgga tcgaagaagg aatcaaggag ctgggcagcc agatcctgaa agagcacccg     2400

gtggaaaaca cgcagctgca gaacgagaag ctctacctgt actatttgca aaatggacgg     2460

gacatgtacg tggaccaaga gctggacatc aatcggttgt ctgattacga cgtggaccac     2520

atcgttccac agtcctttct gaaggatgac tccatcgata acaaggtgtt gactcgcagc     2580

gacaagaaca gagggaagtc agataatgtg ccatcggagg aggtcgtgaa gaagatgaag     2640

aattactggc ggcagctcct gaatgcgaag ctgattaccc agagaaagtt tgacaatctc     2700

actaaagccg agcgcggcgg actctcagag ctggataagg ctggattcat caaacggcag     2760

ctggtcgaga ctcggcagat taccaagcac gtggcgcaga tcctggactc ccgcatgaac     2820

actaaatacg acgagaacga taagctcatc cgggaagtga aggtgattac cctgaaaagc     2880

aaacttgtgt cggactttcg gaaggacttt cagttttaca aagtgagaga aatcaacaac     2940

taccatcacg cgcatgacgc atacctcaac gctgtggtcg gcaccgccct gatcaagaag     3000

taccctaaac ttgaatcgga gtttgtgtac ggagactaca aggtctacga cgtgaggaag     3060

atgatagcca agtccgaaca ggaaatcggg aaagcaactg cgaaatactt cttttactca     3120

aacatcatga acttcttcaa gactgaaatt acgctggcca atggagaaat caggaagagg     3180

ccactgatcg aaactaacgg agaaacgggc gaaatcgtgt gggacaaggg cagggacttc     3240

gcaactgttc gcaaagtgct ctctatgccg caagtcaata ttgtgaagaa aaccgaagtg     3300

caaaccggcg gattttcaaa ggaatcgatc ctcccaaaga gaaatagcga caagctcatt     3360

gcacgcaaga aagactggga cccgaagaag tacggaggat tcgattcgcc gactgtcgca     3420

tactccgtcc tcgtggtggc caaggtggag aagggaaaga gcaagaagct caaatccgtc     3480

aaagagctgc tggggattac catcatggaa cgatcctcgt tcgagaagaa cccgattgat     3540

ttcctggagg cgaagggtta caaggaggtg aagaaggatc tgatcatcaa actgcccaag     3600

tactcactgt tcgaactgga aaatggtcgg aagcgcatgc tggcttcggc cggagaactc     3660

cagaaaggaa atgagctggc cttgcctagc aagtacgtca acttcctcta tcttgcttcg     3720

cactacgaga aactcaaagg gtcaccggaa gataacgaac agaagcagct tttcgtggag     3780

cagcacaagc attatctgga tgaaatcatc gaacaaatct ccgagttttc aaagcgcgtg     3840

atcctcgccg acgccaacct cgacaaagtc ctgtcggcct acaataagca tagagataag     3900

ccgatcagag aacaggccga gaacattatc cacttgttca ccctgactaa cctgggagct     3960

ccagccgcct tcaagtactt cgatactact atcgaccgca aaagatacac gtccaccaag     4020

gaagttctgg acgcgaccct gatccaccaa agcatcactg gactctacga aactaggatc     4080

gatctgtcgc agctgggtgg cgatggtggc ggtggatcct acccatacga cgtgcctgac     4140

tacgcctccg gaggtggtgg ccccaagaag aaacggaagg tgtgatag                  4188


<210>  46
<211>  4459
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 45, Kozak sequence, and 3 UTR of ALB

<400>  46
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatct gccaccatgg ataagaagta ctcgatcggg ctggatatcg gaactaattc      120

cgtgggttgg gcagtgatca cggatgaata caaagtgccg tccaagaagt tcaaggtcct      180

ggggaacacc gatagacaca gcatcaagaa gaatctcatc ggagccctgc tgtttgactc      240

cggcgaaacc gcagaagcga cccggctcaa acgtaccgcg aggcgacgct acacccggcg      300

gaagaatcgc atctgctatc tgcaagaaat cttttcgaac gaaatggcaa aggtggacga      360

cagcttcttc caccgcctgg aagaatcttt cctggtggag gaggacaaga agcatgaacg      420

gcatcctatc tttggaaaca tcgtggacga agtggcgtac cacgaaaagt acccgaccat      480

ctaccatctg cggaagaagt tggttgactc aactgacaag gccgacctca gattgatcta      540

cttggccctc gcccatatga tcaaattccg cggacacttc ctgatcgaag gcgatctgaa      600

ccctgataac tccgacgtgg ataagctgtt cattcaactg gtgcagacct acaaccaact      660

gttcgaagaa aacccaatca atgccagcgg cgtcgatgcc aaggccatcc tgtccgcccg      720

gctgtcgaag tcgcggcgcc tcgaaaacct gatcgcacag ctgccgggag agaagaagaa      780

cggacttttc ggcaacttga tcgctctctc actgggactc actcccaatt tcaagtccaa      840

ttttgacctg gccgaggacg cgaagctgca actctcaaag gacacctacg acgacgactt      900

ggacaatttg ctggcacaaa ttggcgatca gtacgcggat ctgttccttg ccgctaagaa      960

cctttcggac gcaatcttgc tgtccgatat cctgcgcgtg aacaccgaaa taaccaaagc     1020

gccgcttagc gcctcgatga ttaagcggta cgacgagcat caccaggatc tcacgctgct     1080

caaagcgctc gtgagacagc aactgcctga aaagtacaag gagattttct tcgaccagtc     1140

caagaatggg tacgcagggt acatcgatgg aggcgccagc caggaagagt tctataagtt     1200

catcaagcca atcctggaaa agatggacgg aaccgaagaa ctgctggtca agctgaacag     1260

ggaggatctg ctccgcaaac agagaacctt tgacaacgga agcattccac accagatcca     1320

tctgggtgag ctgcacgcca tcttgcggcg ccaggaggac ttttacccat tcctcaagga     1380

caaccgggaa aagatcgaga aaattctgac gttccgcatc ccgtattacg tgggcccact     1440

ggcgcgcggc aattcgcgct tcgcgtggat gactagaaaa tcagaggaaa ccatcactcc     1500

ttggaatttc gaggaagttg tggataaggg agcttcggca caatccttca tcgaacgaat     1560

gaccaacttc gacaagaatc tcccaaacga gaaggtgctt cctaagcaca gcctccttta     1620

cgaatacttc actgtctaca acgaactgac taaagtgaaa tacgttactg aaggaatgag     1680

gaagccggcc tttctgagcg gagaacagaa gaaagcgatt gtcgatctgc tgttcaagac     1740

caaccgcaag gtgaccgtca agcagcttaa agaggactac ttcaagaaga tcgagtgttt     1800

cgactcagtg gaaatcagcg gagtggagga cagattcaac gcttcgctgg gaacctatca     1860

tgatctcctg aagatcatca aggacaagga cttccttgac aacgaggaga acgaggacat     1920

cctggaagat atcgtcctga ccttgaccct tttcgaggat cgcgagatga tcgaggagag     1980

gcttaagacc tacgctcatc tcttcgacga taaggtcatg aaacaactca agcgccgccg     2040

gtacactggt tggggccgcc tctcccgcaa gctgatcaac ggtattcgcg ataaacagag     2100

cggtaaaact atcctggatt tcctcaaatc ggatggcttc gctaatcgta acttcatgca     2160

gttgatccac gacgacagcc tgacctttaa ggaggacatc cagaaagcac aagtgagcgg     2220

acagggagac tcactccatg aacacatcgc gaatctggcc ggttcgccgg cgattaagaa     2280

gggaatcctg caaactgtga aggtggtgga cgagctggtg aaggtcatgg gacggcacaa     2340

accggagaat atcgtgattg aaatggcccg agaaaaccag actacccaga agggccagaa     2400

gaactcccgc gaaaggatga agcggatcga agaaggaatc aaggagctgg gcagccagat     2460

cctgaaagag cacccggtgg aaaacacgca gctgcagaac gagaagctct acctgtacta     2520

tttgcaaaat ggacgggaca tgtacgtgga ccaagagctg gacatcaatc ggttgtctga     2580

ttacgacgtg gaccacatcg ttccacagtc ctttctgaag gatgactcca tcgataacaa     2640

ggtgttgact cgcagcgaca agaacagagg gaagtcagat aatgtgccat cggaggaggt     2700

cgtgaagaag atgaagaatt actggcggca gctcctgaat gcgaagctga ttacccagag     2760

aaagtttgac aatctcacta aagccgagcg cggcggactc tcagagctgg ataaggctgg     2820

attcatcaaa cggcagctgg tcgagactcg gcagattacc aagcacgtgg cgcagatcct     2880

ggactcccgc atgaacacta aatacgacga gaacgataag ctcatccggg aagtgaaggt     2940

gattaccctg aaaagcaaac ttgtgtcgga ctttcggaag gactttcagt tttacaaagt     3000

gagagaaatc aacaactacc atcacgcgca tgacgcatac ctcaacgctg tggtcggcac     3060

cgccctgatc aagaagtacc ctaaacttga atcggagttt gtgtacggag actacaaggt     3120

ctacgacgtg aggaagatga tagccaagtc cgaacaggaa atcgggaaag caactgcgaa     3180

atacttcttt tactcaaaca tcatgaactt cttcaagact gaaattacgc tggccaatgg     3240

agaaatcagg aagaggccac tgatcgaaac taacggagaa acgggcgaaa tcgtgtggga     3300

caagggcagg gacttcgcaa ctgttcgcaa agtgctctct atgccgcaag tcaatattgt     3360

gaagaaaacc gaagtgcaaa ccggcggatt ttcaaaggaa tcgatcctcc caaagagaaa     3420

tagcgacaag ctcattgcac gcaagaaaga ctgggacccg aagaagtacg gaggattcga     3480

ttcgccgact gtcgcatact ccgtcctcgt ggtggccaag gtggagaagg gaaagagcaa     3540

gaagctcaaa tccgtcaaag agctgctggg gattaccatc atggaacgat cctcgttcga     3600

gaagaacccg attgatttcc tggaggcgaa gggttacaag gaggtgaaga aggatctgat     3660

catcaaactg cccaagtact cactgttcga actggaaaat ggtcggaagc gcatgctggc     3720

ttcggccgga gaactccaga aaggaaatga gctggccttg cctagcaagt acgtcaactt     3780

cctctatctt gcttcgcact acgagaaact caaagggtca ccggaagata acgaacagaa     3840

gcagcttttc gtggagcagc acaagcatta tctggatgaa atcatcgaac aaatctccga     3900

gttttcaaag cgcgtgatcc tcgccgacgc caacctcgac aaagtcctgt cggcctacaa     3960

taagcataga gataagccga tcagagaaca ggccgagaac attatccact tgttcaccct     4020

gactaacctg ggagctccag ccgccttcaa gtacttcgat actactatcg accgcaaaag     4080

atacacgtcc accaaggaag ttctggacgc gaccctgatc caccaaagca tcactggact     4140

ctacgaaact aggatcgatc tgtcgcagct gggtggcgat ggtggcggtg gatcctaccc     4200

atacgacgtg cctgactacg cctccggagg tggtggcccc aagaagaaac ggaaggtgtg     4260

atagctagcc atcacattta aaagcatctc agcctaccat gagaataaga gaaagaaaat     4320

gaagatcaat agcttattca tctctttttc tttttcgttg gtgtaaagcc aacaccctgt     4380

ctaaaaaaca taaatttctt taatcatttt gcctcttttc tctgtgcttc aattaataaa     4440

aaatggaaag aacctcgag                                                  4459


<210>  47
<211>  4453
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 45, and 3 UTR of ALB

<400>  47
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatct atggataaga agtactcgat cgggctggat atcggaacta attccgtggg      120

ttgggcagtg atcacggatg aatacaaagt gccgtccaag aagttcaagg tcctggggaa      180

caccgataga cacagcatca agaagaatct catcggagcc ctgctgtttg actccggcga      240

aaccgcagaa gcgacccggc tcaaacgtac cgcgaggcga cgctacaccc ggcggaagaa      300

tcgcatctgc tatctgcaag aaatcttttc gaacgaaatg gcaaaggtgg acgacagctt      360

cttccaccgc ctggaagaat ctttcctggt ggaggaggac aagaagcatg aacggcatcc      420

tatctttgga aacatcgtgg acgaagtggc gtaccacgaa aagtacccga ccatctacca      480

tctgcggaag aagttggttg actcaactga caaggccgac ctcagattga tctacttggc      540

cctcgcccat atgatcaaat tccgcggaca cttcctgatc gaaggcgatc tgaaccctga      600

taactccgac gtggataagc tgttcattca actggtgcag acctacaacc aactgttcga      660

agaaaaccca atcaatgcca gcggcgtcga tgccaaggcc atcctgtccg cccggctgtc      720

gaagtcgcgg cgcctcgaaa acctgatcgc acagctgccg ggagagaaga agaacggact      780

tttcggcaac ttgatcgctc tctcactggg actcactccc aatttcaagt ccaattttga      840

cctggccgag gacgcgaagc tgcaactctc aaaggacacc tacgacgacg acttggacaa      900

tttgctggca caaattggcg atcagtacgc ggatctgttc cttgccgcta agaacctttc      960

ggacgcaatc ttgctgtccg atatcctgcg cgtgaacacc gaaataacca aagcgccgct     1020

tagcgcctcg atgattaagc ggtacgacga gcatcaccag gatctcacgc tgctcaaagc     1080

gctcgtgaga cagcaactgc ctgaaaagta caaggagatt ttcttcgacc agtccaagaa     1140

tgggtacgca gggtacatcg atggaggcgc cagccaggaa gagttctata agttcatcaa     1200

gccaatcctg gaaaagatgg acggaaccga agaactgctg gtcaagctga acagggagga     1260

tctgctccgc aaacagagaa cctttgacaa cggaagcatt ccacaccaga tccatctggg     1320

tgagctgcac gccatcttgc ggcgccagga ggacttttac ccattcctca aggacaaccg     1380

ggaaaagatc gagaaaattc tgacgttccg catcccgtat tacgtgggcc cactggcgcg     1440

cggcaattcg cgcttcgcgt ggatgactag aaaatcagag gaaaccatca ctccttggaa     1500

tttcgaggaa gttgtggata agggagcttc ggcacaatcc ttcatcgaac gaatgaccaa     1560

cttcgacaag aatctcccaa acgagaaggt gcttcctaag cacagcctcc tttacgaata     1620

cttcactgtc tacaacgaac tgactaaagt gaaatacgtt actgaaggaa tgaggaagcc     1680

ggcctttctg agcggagaac agaagaaagc gattgtcgat ctgctgttca agaccaaccg     1740

caaggtgacc gtcaagcagc ttaaagagga ctacttcaag aagatcgagt gtttcgactc     1800

agtggaaatc agcggagtgg aggacagatt caacgcttcg ctgggaacct atcatgatct     1860

cctgaagatc atcaaggaca aggacttcct tgacaacgag gagaacgagg acatcctgga     1920

agatatcgtc ctgaccttga cccttttcga ggatcgcgag atgatcgagg agaggcttaa     1980

gacctacgct catctcttcg acgataaggt catgaaacaa ctcaagcgcc gccggtacac     2040

tggttggggc cgcctctccc gcaagctgat caacggtatt cgcgataaac agagcggtaa     2100

aactatcctg gatttcctca aatcggatgg cttcgctaat cgtaacttca tgcagttgat     2160

ccacgacgac agcctgacct ttaaggagga catccagaaa gcacaagtga gcggacaggg     2220

agactcactc catgaacaca tcgcgaatct ggccggttcg ccggcgatta agaagggaat     2280

cctgcaaact gtgaaggtgg tggacgagct ggtgaaggtc atgggacggc acaaaccgga     2340

gaatatcgtg attgaaatgg cccgagaaaa ccagactacc cagaagggcc agaagaactc     2400

ccgcgaaagg atgaagcgga tcgaagaagg aatcaaggag ctgggcagcc agatcctgaa     2460

agagcacccg gtggaaaaca cgcagctgca gaacgagaag ctctacctgt actatttgca     2520

aaatggacgg gacatgtacg tggaccaaga gctggacatc aatcggttgt ctgattacga     2580

cgtggaccac atcgttccac agtcctttct gaaggatgac tccatcgata acaaggtgtt     2640

gactcgcagc gacaagaaca gagggaagtc agataatgtg ccatcggagg aggtcgtgaa     2700

gaagatgaag aattactggc ggcagctcct gaatgcgaag ctgattaccc agagaaagtt     2760

tgacaatctc actaaagccg agcgcggcgg actctcagag ctggataagg ctggattcat     2820

caaacggcag ctggtcgaga ctcggcagat taccaagcac gtggcgcaga tcctggactc     2880

ccgcatgaac actaaatacg acgagaacga taagctcatc cgggaagtga aggtgattac     2940

cctgaaaagc aaacttgtgt cggactttcg gaaggacttt cagttttaca aagtgagaga     3000

aatcaacaac taccatcacg cgcatgacgc atacctcaac gctgtggtcg gcaccgccct     3060

gatcaagaag taccctaaac ttgaatcgga gtttgtgtac ggagactaca aggtctacga     3120

cgtgaggaag atgatagcca agtccgaaca ggaaatcggg aaagcaactg cgaaatactt     3180

cttttactca aacatcatga acttcttcaa gactgaaatt acgctggcca atggagaaat     3240

caggaagagg ccactgatcg aaactaacgg agaaacgggc gaaatcgtgt gggacaaggg     3300

cagggacttc gcaactgttc gcaaagtgct ctctatgccg caagtcaata ttgtgaagaa     3360

aaccgaagtg caaaccggcg gattttcaaa ggaatcgatc ctcccaaaga gaaatagcga     3420

caagctcatt gcacgcaaga aagactggga cccgaagaag tacggaggat tcgattcgcc     3480

gactgtcgca tactccgtcc tcgtggtggc caaggtggag aagggaaaga gcaagaagct     3540

caaatccgtc aaagagctgc tggggattac catcatggaa cgatcctcgt tcgagaagaa     3600

cccgattgat ttcctggagg cgaagggtta caaggaggtg aagaaggatc tgatcatcaa     3660

actgcccaag tactcactgt tcgaactgga aaatggtcgg aagcgcatgc tggcttcggc     3720

cggagaactc cagaaaggaa atgagctggc cttgcctagc aagtacgtca acttcctcta     3780

tcttgcttcg cactacgaga aactcaaagg gtcaccggaa gataacgaac agaagcagct     3840

tttcgtggag cagcacaagc attatctgga tgaaatcatc gaacaaatct ccgagttttc     3900

aaagcgcgtg atcctcgccg acgccaacct cgacaaagtc ctgtcggcct acaataagca     3960

tagagataag ccgatcagag aacaggccga gaacattatc cacttgttca ccctgactaa     4020

cctgggagct ccagccgcct tcaagtactt cgatactact atcgaccgca aaagatacac     4080

gtccaccaag gaagttctgg acgcgaccct gatccaccaa agcatcactg gactctacga     4140

aactaggatc gatctgtcgc agctgggtgg cgatggtggc ggtggatcct acccatacga     4200

cgtgcctgac tacgcctccg gaggtggtgg ccccaagaag aaacggaagg tgtgatagct     4260

agccatcaca tttaaaagca tctcagccta ccatgagaat aagagaaaga aaatgaagat     4320

caatagctta ttcatctctt tttctttttc gttggtgtaa agccaacacc ctgtctaaaa     4380

aacataaatt tctttaatca ttttgcctct tttctctgtg cttcaattaa taaaaaatgg     4440

aaagaacctc gag                                                        4453


<210>  48
<211>  4403
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript comprising Cas9 ORF using codons with 
       generally high expression in humans

<400>  48
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc atgcctaaga aaaagcggaa ggtcgacggg gataagaagt actcaatcgg      120

gctggatatc ggaactaatt ccgtgggttg ggcagtgatc acggatgaat acaaagtgcc      180

gtccaagaag ttcaaggtcc tggggaacac cgatagacac agcatcaaga aaaatctcat      240

cggagccctg ctgtttgact ccggcgaaac cgcagaagcg acccggctca aacgtaccgc      300

gaggcgacgc tacacccggc ggaagaatcg catctgctat ctgcaagaga tcttttcgaa      360

cgaaatggca aaggtcgacg acagcttctt ccaccgcctg gaagaatctt tcctggtgga      420

ggaggacaag aagcatgaac ggcatcctat ctttggaaac atcgtcgacg aagtggcgta      480

ccacgaaaag tacccgacca tctaccatct gcggaagaag ttggttgact caactgacaa      540

ggccgacctc agattgatct acttggccct cgcccatatg atcaaattcc gcggacactt      600

cctgatcgaa ggcgatctga accctgataa ctccgacgtg gataagcttt tcattcaact      660

ggtgcagacc tacaaccaac tgttcgaaga aaacccaatc aatgctagcg gcgtcgatgc      720

caaggccatc ctgtccgccc ggctgtcgaa gtcgcggcgc ctcgaaaacc tgatcgcaca      780

gctgccggga gagaaaaaga acggactttt cggcaacttg atcgctctct cactgggact      840

cactcccaat ttcaagtcca attttgacct ggccgaggac gcgaagctgc aactctcaaa      900

ggacacctac gacgacgact tggacaattt gctggcacaa attggcgatc agtacgcgga      960

tctgttcctt gccgctaaga acctttcgga cgcaatcttg ctgtccgata tcctgcgcgt     1020

gaacaccgaa ataaccaaag cgccgcttag cgcctcgatg attaagcggt acgacgagca     1080

tcaccaggat ctcacgctgc tcaaagcgct cgtgagacag caactgcctg aaaagtacaa     1140

ggagatcttc ttcgaccagt ccaagaatgg gtacgcaggg tacatcgatg gaggcgctag     1200

ccaggaagag ttctataagt tcatcaagcc aatcctggaa aagatggacg gaaccgaaga     1260

actgctggtc aagctgaaca gggaggatct gctccggaaa cagagaacct ttgacaacgg     1320

atccattccc caccagatcc atctgggtga gctgcacgcc atcttgcggc gccaggagga     1380

cttttaccca ttcctcaagg acaaccggga aaagatcgag aaaattctga cgttccgcat     1440

cccgtattac gtgggcccac tggcgcgcgg caattcgcgc ttcgcgtgga tgactagaaa     1500

atcagaggaa accatcactc cttggaattt cgaggaagtt gtggataagg gagcttcggc     1560

acaaagcttc atcgaacgaa tgaccaactt cgacaagaat ctcccaaacg agaaggtgct     1620

tcctaagcac agcctccttt acgaatactt cactgtctac aacgaactga ctaaagtgaa     1680

atacgttact gaaggaatga ggaagccggc ctttctgtcc ggagaacaga agaaagcaat     1740

tgtcgatctg ctgttcaaga ccaaccgcaa ggtgaccgtc aagcagctta aagaggacta     1800

cttcaagaag atcgagtgtt tcgactcagt ggaaatcagc ggggtggagg acagattcaa     1860

cgcttcgctg ggaacctatc atgatctcct gaagatcatc aaggacaagg acttccttga     1920

caacgaggag aacgaggaca tcctggaaga tatcgtcctg accttgaccc ttttcgagga     1980

tcgcgagatg atcgaggaga ggcttaagac ctacgctcat ctcttcgacg ataaggtcat     2040

gaaacaactc aagcgccgcc ggtacactgg ttggggccgc ctctcccgca agctgatcaa     2100

cggtattcgc gataaacaga gcggtaaaac tatcctggat ttcctcaaat cggatggctt     2160

cgctaatcgt aacttcatgc aattgatcca cgacgacagc ctgaccttta aggaggacat     2220

ccaaaaagca caagtgtccg gacagggaga ctcactccat gaacacatcg cgaatctggc     2280

cggttcgccg gcgattaaga agggaattct gcaaactgtg aaggtggtcg acgagctggt     2340

gaaggtcatg ggacggcaca aaccggagaa tatcgtgatt gaaatggccc gagaaaacca     2400

gactacccag aagggccaga aaaactcccg cgaaaggatg aagcggatcg aagaaggaat     2460

caaggagctg ggcagccaga tcctgaaaga gcacccggtg gaaaacacgc agctgcagaa     2520

cgagaagctc tacctgtact atttgcaaaa tggacgggac atgtacgtgg accaagagct     2580

ggacatcaat cggttgtctg attacgacgt ggaccacatc gttccacagt cctttctgaa     2640

ggatgactcg atcgataaca aggtgttgac tcgcagcgac aagaacagag ggaagtcaga     2700

taatgtgcca tcggaggagg tcgtgaagaa gatgaagaat tactggcggc agctcctgaa     2760

tgcgaagctg attacccaga gaaagtttga caatctcact aaagccgagc gcggcggact     2820

ctcagagctg gataaggctg gattcatcaa acggcagctg gtcgagactc ggcagattac     2880

caagcacgtg gcgcagatct tggactcccg catgaacact aaatacgacg agaacgataa     2940

gctcatccgg gaagtgaagg tgattaccct gaaaagcaaa cttgtgtcgg actttcggaa     3000

ggactttcag ttttacaaag tgagagaaat caacaactac catcacgcgc atgacgcata     3060

cctcaacgct gtggtcggta ccgccctgat caaaaagtac cctaaacttg aatcggagtt     3120

tgtgtacgga gactacaagg tctacgacgt gaggaagatg atagccaagt ccgaacagga     3180

aatcgggaaa gcaactgcga aatacttctt ttactcaaac atcatgaact ttttcaagac     3240

tgaaattacg ctggccaatg gagaaatcag gaagaggcca ctgatcgaaa ctaacggaga     3300

aacgggcgaa atcgtgtggg acaagggcag ggacttcgca actgttcgca aagtgctctc     3360

tatgccgcaa gtcaatattg tgaagaaaac cgaagtgcaa accggcggat tttcaaagga     3420

atcgatcctc ccaaagagaa atagcgacaa gctcattgca cgcaagaaag actgggaccc     3480

gaagaagtac ggaggattcg attcgccgac tgtcgcatac tccgtcctcg tggtggccaa     3540

ggtggagaag ggaaagagca aaaagctcaa atccgtcaaa gagctgctgg ggattaccat     3600

catggaacga tcctcgttcg agaagaaccc gattgatttc ctcgaggcga agggttacaa     3660

ggaggtgaag aaggatctga tcatcaaact ccccaagtac tcactgttcg aactggaaaa     3720

tggtcggaag cgcatgctgg cttcggccgg agaactccaa aaaggaaatg agctggcctt     3780

gcctagcaag tacgtcaact tcctctatct tgcttcgcac tacgaaaaac tcaaagggtc     3840

accggaagat aacgaacaga agcagctttt cgtggagcag cacaagcatt atctggatga     3900

aatcatcgaa caaatctccg agttttcaaa gcgcgtgatc ctcgccgacg ccaacctcga     3960

caaagtcctg tcggcctaca ataagcatag agataagccg atcagagaac aggccgagaa     4020

cattatccac ttgttcaccc tgactaacct gggagcccca gccgccttca agtacttcga     4080

tactactatc gatcgcaaaa gatacacgtc caccaaggaa gttctggacg cgaccctgat     4140

ccaccaaagc atcactggac tctacgaaac taggatcgat ctgtcgcagc tgggtggcga     4200

ttgatagtct agccatcaca tttaaaagca tctcagccta ccatgagaat aagagaaaga     4260

aaatgaagat caatagctta ttcatctctt tttctttttc gttggtgtaa agccaacacc     4320

ctgtctaaaa aacataaatt tctttaatca ttttgcctct tttctctgtg cttcaattaa     4380

taaaaaatgg aaagaacctc gag                                             4403


<210>  49
<211>  4409
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript comprising Kozak sequence with Cas9 
       ORF using codons with generally high expression in humans

<400>  49
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgc ctaagaaaaa gcggaaggtc gacggggata agaagtactc      120

aatcgggctg gatatcggaa ctaattccgt gggttgggca gtgatcacgg atgaatacaa      180

agtgccgtcc aagaagttca aggtcctggg gaacaccgat agacacagca tcaagaaaaa      240

tctcatcgga gccctgctgt ttgactccgg cgaaaccgca gaagcgaccc ggctcaaacg      300

taccgcgagg cgacgctaca cccggcggaa gaatcgcatc tgctatctgc aagagatctt      360

ttcgaacgaa atggcaaagg tcgacgacag cttcttccac cgcctggaag aatctttcct      420

ggtggaggag gacaagaagc atgaacggca tcctatcttt ggaaacatcg tcgacgaagt      480

ggcgtaccac gaaaagtacc cgaccatcta ccatctgcgg aagaagttgg ttgactcaac      540

tgacaaggcc gacctcagat tgatctactt ggccctcgcc catatgatca aattccgcgg      600

acacttcctg atcgaaggcg atctgaaccc tgataactcc gacgtggata agcttttcat      660

tcaactggtg cagacctaca accaactgtt cgaagaaaac ccaatcaatg ctagcggcgt      720

cgatgccaag gccatcctgt ccgcccggct gtcgaagtcg cggcgcctcg aaaacctgat      780

cgcacagctg ccgggagaga aaaagaacgg acttttcggc aacttgatcg ctctctcact      840

gggactcact cccaatttca agtccaattt tgacctggcc gaggacgcga agctgcaact      900

ctcaaaggac acctacgacg acgacttgga caatttgctg gcacaaattg gcgatcagta      960

cgcggatctg ttccttgccg ctaagaacct ttcggacgca atcttgctgt ccgatatcct     1020

gcgcgtgaac accgaaataa ccaaagcgcc gcttagcgcc tcgatgatta agcggtacga     1080

cgagcatcac caggatctca cgctgctcaa agcgctcgtg agacagcaac tgcctgaaaa     1140

gtacaaggag atcttcttcg accagtccaa gaatgggtac gcagggtaca tcgatggagg     1200

cgctagccag gaagagttct ataagttcat caagccaatc ctggaaaaga tggacggaac     1260

cgaagaactg ctggtcaagc tgaacaggga ggatctgctc cggaaacaga gaacctttga     1320

caacggatcc attccccacc agatccatct gggtgagctg cacgccatct tgcggcgcca     1380

ggaggacttt tacccattcc tcaaggacaa ccgggaaaag atcgagaaaa ttctgacgtt     1440

ccgcatcccg tattacgtgg gcccactggc gcgcggcaat tcgcgcttcg cgtggatgac     1500

tagaaaatca gaggaaacca tcactccttg gaatttcgag gaagttgtgg ataagggagc     1560

ttcggcacaa agcttcatcg aacgaatgac caacttcgac aagaatctcc caaacgagaa     1620

ggtgcttcct aagcacagcc tcctttacga atacttcact gtctacaacg aactgactaa     1680

agtgaaatac gttactgaag gaatgaggaa gccggccttt ctgtccggag aacagaagaa     1740

agcaattgtc gatctgctgt tcaagaccaa ccgcaaggtg accgtcaagc agcttaaaga     1800

ggactacttc aagaagatcg agtgtttcga ctcagtggaa atcagcgggg tggaggacag     1860

attcaacgct tcgctgggaa cctatcatga tctcctgaag atcatcaagg acaaggactt     1920

ccttgacaac gaggagaacg aggacatcct ggaagatatc gtcctgacct tgaccctttt     1980

cgaggatcgc gagatgatcg aggagaggct taagacctac gctcatctct tcgacgataa     2040

ggtcatgaaa caactcaagc gccgccggta cactggttgg ggccgcctct cccgcaagct     2100

gatcaacggt attcgcgata aacagagcgg taaaactatc ctggatttcc tcaaatcgga     2160

tggcttcgct aatcgtaact tcatgcaatt gatccacgac gacagcctga cctttaagga     2220

ggacatccaa aaagcacaag tgtccggaca gggagactca ctccatgaac acatcgcgaa     2280

tctggccggt tcgccggcga ttaagaaggg aattctgcaa actgtgaagg tggtcgacga     2340

gctggtgaag gtcatgggac ggcacaaacc ggagaatatc gtgattgaaa tggcccgaga     2400

aaaccagact acccagaagg gccagaaaaa ctcccgcgaa aggatgaagc ggatcgaaga     2460

aggaatcaag gagctgggca gccagatcct gaaagagcac ccggtggaaa acacgcagct     2520

gcagaacgag aagctctacc tgtactattt gcaaaatgga cgggacatgt acgtggacca     2580

agagctggac atcaatcggt tgtctgatta cgacgtggac cacatcgttc cacagtcctt     2640

tctgaaggat gactcgatcg ataacaaggt gttgactcgc agcgacaaga acagagggaa     2700

gtcagataat gtgccatcgg aggaggtcgt gaagaagatg aagaattact ggcggcagct     2760

cctgaatgcg aagctgatta cccagagaaa gtttgacaat ctcactaaag ccgagcgcgg     2820

cggactctca gagctggata aggctggatt catcaaacgg cagctggtcg agactcggca     2880

gattaccaag cacgtggcgc agatcttgga ctcccgcatg aacactaaat acgacgagaa     2940

cgataagctc atccgggaag tgaaggtgat taccctgaaa agcaaacttg tgtcggactt     3000

tcggaaggac tttcagtttt acaaagtgag agaaatcaac aactaccatc acgcgcatga     3060

cgcatacctc aacgctgtgg tcggtaccgc cctgatcaaa aagtacccta aacttgaatc     3120

ggagtttgtg tacggagact acaaggtcta cgacgtgagg aagatgatag ccaagtccga     3180

acaggaaatc gggaaagcaa ctgcgaaata cttcttttac tcaaacatca tgaacttttt     3240

caagactgaa attacgctgg ccaatggaga aatcaggaag aggccactga tcgaaactaa     3300

cggagaaacg ggcgaaatcg tgtgggacaa gggcagggac ttcgcaactg ttcgcaaagt     3360

gctctctatg ccgcaagtca atattgtgaa gaaaaccgaa gtgcaaaccg gcggattttc     3420

aaaggaatcg atcctcccaa agagaaatag cgacaagctc attgcacgca agaaagactg     3480

ggacccgaag aagtacggag gattcgattc gccgactgtc gcatactccg tcctcgtggt     3540

ggccaaggtg gagaagggaa agagcaaaaa gctcaaatcc gtcaaagagc tgctggggat     3600

taccatcatg gaacgatcct cgttcgagaa gaacccgatt gatttcctcg aggcgaaggg     3660

ttacaaggag gtgaagaagg atctgatcat caaactcccc aagtactcac tgttcgaact     3720

ggaaaatggt cggaagcgca tgctggcttc ggccggagaa ctccaaaaag gaaatgagct     3780

ggccttgcct agcaagtacg tcaacttcct ctatcttgct tcgcactacg aaaaactcaa     3840

agggtcaccg gaagataacg aacagaagca gcttttcgtg gagcagcaca agcattatct     3900

ggatgaaatc atcgaacaaa tctccgagtt ttcaaagcgc gtgatcctcg ccgacgccaa     3960

cctcgacaaa gtcctgtcgg cctacaataa gcatagagat aagccgatca gagaacaggc     4020

cgagaacatt atccacttgt tcaccctgac taacctggga gccccagccg ccttcaagta     4080

cttcgatact actatcgatc gcaaaagata cacgtccacc aaggaagttc tggacgcgac     4140

cctgatccac caaagcatca ctggactcta cgaaactagg atcgatctgt cgcagctggg     4200

tggcgattga tagtctagcc atcacattta aaagcatctc agcctaccat gagaataaga     4260

gaaagaaaat gaagatcaat agcttattca tctctttttc tttttcgttg gtgtaaagcc     4320

aacaccctgt ctaaaaaaca taaatttctt taatcatttt gcctcttttc tctgtgcttc     4380

aattaataaa aaatggaaag aacctcgag                                       4409


<210>  50
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF with splice junctions removed; 12.75% U 
       content

<400>  50
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccaccgg      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca aaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcccga agaagaagag aaaggtctag     4140


<210>  51
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 50, Kozak sequence, and 3 UTR of ALB

<400>  51
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc caccggctgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcaaaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  52
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF with minimal uridine codons frequently used 
       in humans in general; 12.75% U content

<400>  52
atggacaaga agtacagcat cggcctggac atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgacaga      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccaccagac tgaagagaac cgccagaaga agatacacca gaagaaagaa cagaatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agagacaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag      420

aagctggtgg acagcaccga caaggccgac ctgagactga tctacctggc cctggcccac      480

atgatcaagt tcagaggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg ccagactgag caagagcaga      660

agactggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgaga     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga acagagagga cctgctgaga     1200

aagcagagaa ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctga gaagacagga ggacttctac cccttcctga aggacaacag agagaagatc     1320

gagaagatcc tgaccttcag aatcccctac tacgtgggcc ccctggccag aggcaacagc     1380

agattcgcct ggatgaccag aaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgaga gaatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgagaaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaacag aaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggacagatt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggacagagag atgatcgagg agagactgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagagaa gaagatacac cggctggggc     1980

agactgagca gaaagctgat caacggcatc agagacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggcagac acaagcccga gaacatcgtg     2280

atcgagatgg ccagagagaa ccagaccacc cagaagggcc agaagaacag cagagagaga     2340

atgaagagaa tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggcaga     2460

gacatgtacg tggaccagga gctggacatc aacagactga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gaccagaagc     2580

gacaagaaca gaggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactgga gacagctgct gaacgccaag ctgatcaccc agagaaagtt cgacaacctg     2700

accaaggccg agagaggcgg cctgagcgag ctggacaagg ccggcttcat caagagacag     2760

ctggtggaga ccagacagat caccaagcac gtggcccaga tcctggacag cagaatgaac     2820

accaagtacg acgagaacga caagctgatc agagaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttcag aaaggacttc cagttctaca aggtgagaga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgagaaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat cagaaagaga     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg cagagacttc     3240

gccaccgtga gaaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaaga gaaacagcga caagctgatc     3360

gccagaaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag agaagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggcaga aagagaatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagagagtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca cagagacaag     3900

cccatcagag agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgacagaa agagatacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gaccagaatc     4080

gacctgagcc agctgggcgg cgacggcggc ggcagcccca agaagaagag aaaggtgtga     4140


<210>  53
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 52, Kozak sequence, and 3 UTR of ALB

<400>  53
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcggc ctggacatcg gcaccaacag      120

cgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaagt tcaaggtgct      180

gggcaacacc gacagacaca gcatcaagaa gaacctgatc ggcgccctgc tgttcgacag      240

cggcgagacc gccgaggcca ccagactgaa gagaaccgcc agaagaagat acaccagaag      300

aaagaacaga atctgctacc tgcaggagat cttcagcaac gagatggcca aggtggacga      360

cagcttcttc cacagactgg aggagagctt cctggtggag gaggacaaga agcacgagag      420

acaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat      480

ctaccacctg agaaagaagc tggtggacag caccgacaag gccgacctga gactgatcta      540

cctggccctg gcccacatga tcaagttcag aggccacttc ctgatcgagg gcgacctgaa      600

ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct      660

gttcgaggag aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgagcgccag      720

actgagcaag agcagaagac tggagaacct gatcgcccag ctgcccggcg agaagaagaa      780

cggcctgttc ggcaacctga tcgccctgag cctgggcctg acccccaact tcaagagcaa      840

cttcgacctg gccgaggacg ccaagctgca gctgagcaag gacacctacg acgacgacct      900

ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgttcctgg ccgccaagaa      960

cctgagcgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc     1020

ccccctgagc gccagcatga tcaagagata cgacgagcac caccaggacc tgaccctgct     1080

gaaggccctg gtgagacagc agctgcccga gaagtacaag gagatcttct tcgaccagag     1140

caagaacggc tacgccggct acatcgacgg cggcgccagc caggaggagt tctacaagtt     1200

catcaagccc atcctggaga agatggacgg caccgaggag ctgctggtga agctgaacag     1260

agaggacctg ctgagaaagc agagaacctt cgacaacggc agcatccccc accagatcca     1320

cctgggcgag ctgcacgcca tcctgagaag acaggaggac ttctacccct tcctgaagga     1380

caacagagag aagatcgaga agatcctgac cttcagaatc ccctactacg tgggccccct     1440

ggccagaggc aacagcagat tcgcctggat gaccagaaag agcgaggaga ccatcacccc     1500

ctggaacttc gaggaggtgg tggacaaggg cgccagcgcc cagagcttca tcgagagaat     1560

gaccaacttc gacaagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta     1620

cgagtacttc accgtgtaca acgagctgac caaggtgaag tacgtgaccg agggcatgag     1680

aaagcccgcc ttcctgagcg gcgagcagaa gaaggccatc gtggacctgc tgttcaagac     1740

caacagaaag gtgaccgtga agcagctgaa ggaggactac ttcaagaaga tcgagtgctt     1800

cgacagcgtg gagatcagcg gcgtggagga cagattcaac gccagcctgg gcacctacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaggaga acgaggacat     1920

cctggaggac atcgtgctga ccctgaccct gttcgaggac agagagatga tcgaggagag     1980

actgaagacc tacgcccacc tgttcgacga caaggtgatg aagcagctga agagaagaag     2040

atacaccggc tggggcagac tgagcagaaa gctgatcaac ggcatcagag acaagcagag     2100

cggcaagacc atcctggact tcctgaagag cgacggcttc gccaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgaccttcaa ggaggacatc cagaaggccc aggtgagcgg     2220

ccagggcgac agcctgcacg agcacatcgc caacctggcc ggcagccccg ccatcaagaa     2280

gggcatcctg cagaccgtga aggtggtgga cgagctggtg aaggtgatgg gcagacacaa     2340

gcccgagaac atcgtgatcg agatggccag agagaaccag accacccaga agggccagaa     2400

gaacagcaga gagagaatga agagaatcga ggagggcatc aaggagctgg gcagccagat     2460

cctgaaggag caccccgtgg agaacaccca gctgcagaac gagaagctgt acctgtacta     2520

cctgcagaac ggcagagaca tgtacgtgga ccaggagctg gacatcaaca gactgagcga     2580

ctacgacgtg gaccacatcg tgccccagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtgctgacc agaagcgaca agaacagagg caagagcgac aacgtgccca gcgaggaggt     2700

ggtgaagaag atgaagaact actggagaca gctgctgaac gccaagctga tcacccagag     2760

aaagttcgac aacctgacca aggccgagag aggcggcctg agcgagctgg acaaggccgg     2820

cttcatcaag agacagctgg tggagaccag acagatcacc aagcacgtgg cccagatcct     2880

ggacagcaga atgaacacca agtacgacga gaacgacaag ctgatcagag aggtgaaggt     2940

gatcaccctg aagagcaagc tggtgagcga cttcagaaag gacttccagt tctacaaggt     3000

gagagagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tggtgggcac     3060

cgccctgatc aagaagtacc ccaagctgga gagcgagttc gtgtacggcg actacaaggt     3120

gtacgacgtg agaaagatga tcgccaagag cgagcaggag atcggcaagg ccaccgccaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagacc gagatcaccc tggccaacgg     3240

cgagatcaga aagagacccc tgatcgagac caacggcgag accggcgaga tcgtgtggga     3300

caagggcaga gacttcgcca ccgtgagaaa ggtgctgagc atgccccagg tgaacatcgt     3360

gaagaagacc gaggtgcaga ccggcggctt cagcaaggag agcatcctgc ccaagagaaa     3420

cagcgacaag ctgatcgcca gaaagaagga ctgggacccc aagaagtacg gcggcttcga     3480

cagccccacc gtggcctaca gcgtgctggt ggtggccaag gtggagaagg gcaagagcaa     3540

gaagctgaag agcgtgaagg agctgctggg catcaccatc atggagagaa gcagcttcga     3600

gaagaacccc atcgacttcc tggaggccaa gggctacaag gaggtgaaga aggacctgat     3660

catcaagctg cccaagtaca gcctgttcga gctggagaac ggcagaaaga gaatgctggc     3720

cagcgccggc gagctgcaga agggcaacga gctggccctg cccagcaagt acgtgaactt     3780

cctgtacctg gccagccact acgagaagct gaagggcagc cccgaggaca acgagcagaa     3840

gcagctgttc gtggagcagc acaagcacta cctggacgag atcatcgagc agatcagcga     3900

gttcagcaag agagtgatcc tggccgacgc caacctggac aaggtgctga gcgcctacaa     3960

caagcacaga gacaagccca tcagagagca ggccgagaac atcatccacc tgttcaccct     4020

gaccaacctg ggcgcccccg ccgccttcaa gtacttcgac accaccatcg acagaaagag     4080

atacaccagc accaaggagg tgctggacgc caccctgatc caccagagca tcaccggcct     4140

gtacgagacc agaatcgacc tgagccagct gggcggcgac ggcggcggca gccccaagaa     4200

gaagagaaag gtgtgactag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  54
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF with minimal uridine codons infrequently used
       in humans in general; 12.75% U content

<400>  54
atggacaaaa aatacagcat agggctagac atagggacga acagcgtagg gtgggcggta       60

ataacggacg aatacaaagt accgagcaaa aaattcaaag tactagggaa cacggaccga      120

cacagcataa aaaaaaacct aataggggcg ctactattcg acagcgggga aacggcggaa      180

gcgacgcgac taaaacgaac ggcgcgacga cgatacacgc gacgaaaaaa ccgaatatgc      240

tacctacaag aaatattcag caacgaaatg gcgaaagtag acgacagctt cttccaccga      300

ctagaagaaa gcttcctagt agaagaagac aaaaaacacg aacgacaccc gatattcggg      360

aacatagtag acgaagtagc gtaccacgaa aaatacccga cgatatacca cctacgaaaa      420

aaactagtag acagcacgga caaagcggac ctacgactaa tatacctagc gctagcgcac      480

atgataaaat tccgagggca cttcctaata gaaggggacc taaacccgga caacagcgac      540

gtagacaaac tattcataca actagtacaa acgtacaacc aactattcga agaaaacccg      600

ataaacgcga gcggggtaga cgcgaaagcg atactaagcg cgcgactaag caaaagccga      660

cgactagaaa acctaatagc gcaactaccg ggggaaaaaa aaaacgggct attcgggaac      720

ctaatagcgc taagcctagg gctaacgccg aacttcaaaa gcaacttcga cctagcggaa      780

gacgcgaaac tacaactaag caaagacacg tacgacgacg acctagacaa cctactagcg      840

caaatagggg accaatacgc ggacctattc ctagcggcga aaaacctaag cgacgcgata      900

ctactaagcg acatactacg agtaaacacg gaaataacga aagcgccgct aagcgcgagc      960

atgataaaac gatacgacga acaccaccaa gacctaacgc tactaaaagc gctagtacga     1020

caacaactac cggaaaaata caaagaaata ttcttcgacc aaagcaaaaa cgggtacgcg     1080

gggtacatag acgggggggc gagccaagaa gaattctaca aattcataaa accgatacta     1140

gaaaaaatgg acgggacgga agaactacta gtaaaactaa accgagaaga cctactacga     1200

aaacaacgaa cgttcgacaa cgggagcata ccgcaccaaa tacacctagg ggaactacac     1260

gcgatactac gacgacaaga agacttctac ccgttcctaa aagacaaccg agaaaaaata     1320

gaaaaaatac taacgttccg aataccgtac tacgtagggc cgctagcgcg agggaacagc     1380

cgattcgcgt ggatgacgcg aaaaagcgaa gaaacgataa cgccgtggaa cttcgaagaa     1440

gtagtagaca aaggggcgag cgcgcaaagc ttcatagaac gaatgacgaa cttcgacaaa     1500

aacctaccga acgaaaaagt actaccgaaa cacagcctac tatacgaata cttcacggta     1560

tacaacgaac taacgaaagt aaaatacgta acggaaggga tgcgaaaacc ggcgttccta     1620

agcggggaac aaaaaaaagc gatagtagac ctactattca aaacgaaccg aaaagtaacg     1680

gtaaaacaac taaaagaaga ctacttcaaa aaaatagaat gcttcgacag cgtagaaata     1740

agcggggtag aagaccgatt caacgcgagc ctagggacgt accacgacct actaaaaata     1800

ataaaagaca aagacttcct agacaacgaa gaaaacgaag acatactaga agacatagta     1860

ctaacgctaa cgctattcga agaccgagaa atgatagaag aacgactaaa aacgtacgcg     1920

cacctattcg acgacaaagt aatgaaacaa ctaaaacgac gacgatacac ggggtggggg     1980

cgactaagcc gaaaactaat aaacgggata cgagacaaac aaagcgggaa aacgatacta     2040

gacttcctaa aaagcgacgg gttcgcgaac cgaaacttca tgcaactaat acacgacgac     2100

agcctaacgt tcaaagaaga catacaaaaa gcgcaagtaa gcgggcaagg ggacagccta     2160

cacgaacaca tagcgaacct agcggggagc ccggcgataa aaaaagggat actacaaacg     2220

gtaaaagtag tagacgaact agtaaaagta atggggcgac acaaaccgga aaacatagta     2280

atagaaatgg cgcgagaaaa ccaaacgacg caaaaagggc aaaaaaacag ccgagaacga     2340

atgaaacgaa tagaagaagg gataaaagaa ctagggagcc aaatactaaa agaacacccg     2400

gtagaaaaca cgcaactaca aaacgaaaaa ctatacctat actacctaca aaacgggcga     2460

gacatgtacg tagaccaaga actagacata aaccgactaa gcgactacga cgtagaccac     2520

atagtaccgc aaagcttcct aaaagacgac agcatagaca acaaagtact aacgcgaagc     2580

gacaaaaacc gagggaaaag cgacaacgta ccgagcgaag aagtagtaaa aaaaatgaaa     2640

aactactggc gacaactact aaacgcgaaa ctaataacgc aacgaaaatt cgacaaccta     2700

acgaaagcgg aacgaggggg gctaagcgaa ctagacaaag cggggttcat aaaacgacaa     2760

ctagtagaaa cgcgacaaat aacgaaacac gtagcgcaaa tactagacag ccgaatgaac     2820

acgaaatacg acgaaaacga caaactaata cgagaagtaa aagtaataac gctaaaaagc     2880

aaactagtaa gcgacttccg aaaagacttc caattctaca aagtacgaga aataaacaac     2940

taccaccacg cgcacgacgc gtacctaaac gcggtagtag ggacggcgct aataaaaaaa     3000

tacccgaaac tagaaagcga attcgtatac ggggactaca aagtatacga cgtacgaaaa     3060

atgatagcga aaagcgaaca agaaataggg aaagcgacgg cgaaatactt cttctacagc     3120

aacataatga acttcttcaa aacggaaata acgctagcga acggggaaat acgaaaacga     3180

ccgctaatag aaacgaacgg ggaaacgggg gaaatagtat gggacaaagg gcgagacttc     3240

gcgacggtac gaaaagtact aagcatgccg caagtaaaca tagtaaaaaa aacggaagta     3300

caaacggggg ggttcagcaa agaaagcata ctaccgaaac gaaacagcga caaactaata     3360

gcgcgaaaaa aagactggga cccgaaaaaa tacggggggt tcgacagccc gacggtagcg     3420

tacagcgtac tagtagtagc gaaagtagaa aaagggaaaa gcaaaaaact aaaaagcgta     3480

aaagaactac tagggataac gataatggaa cgaagcagct tcgaaaaaaa cccgatagac     3540

ttcctagaag cgaaagggta caaagaagta aaaaaagacc taataataaa actaccgaaa     3600

tacagcctat tcgaactaga aaacgggcga aaacgaatgc tagcgagcgc gggggaacta     3660

caaaaaggga acgaactagc gctaccgagc aaatacgtaa acttcctata cctagcgagc     3720

cactacgaaa aactaaaagg gagcccggaa gacaacgaac aaaaacaact attcgtagaa     3780

caacacaaac actacctaga cgaaataata gaacaaataa gcgaattcag caaacgagta     3840

atactagcgg acgcgaacct agacaaagta ctaagcgcgt acaacaaaca ccgagacaaa     3900

ccgatacgag aacaagcgga aaacataata cacctattca cgctaacgaa cctaggggcg     3960

ccggcggcgt tcaaatactt cgacacgacg atagaccgaa aacgatacac gagcacgaaa     4020

gaagtactag acgcgacgct aatacaccaa agcataacgg ggctatacga aacgcgaata     4080

gacctaagcc aactaggggg ggacgggggg gggagcccga aaaaaaaacg aaaagtatga     4140


<210>  55
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR of HSD, ORF corresponding 
       to SEQ ID NO: 54, Kozak sequence, and 3 UTR of ALB

<400>  55
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaaaaaata cagcataggg ctagacatag ggacgaacag      120

cgtagggtgg gcggtaataa cggacgaata caaagtaccg agcaaaaaat tcaaagtact      180

agggaacacg gaccgacaca gcataaaaaa aaacctaata ggggcgctac tattcgacag      240

cggggaaacg gcggaagcga cgcgactaaa acgaacggcg cgacgacgat acacgcgacg      300

aaaaaaccga atatgctacc tacaagaaat attcagcaac gaaatggcga aagtagacga      360

cagcttcttc caccgactag aagaaagctt cctagtagaa gaagacaaaa aacacgaacg      420

acacccgata ttcgggaaca tagtagacga agtagcgtac cacgaaaaat acccgacgat      480

ataccaccta cgaaaaaaac tagtagacag cacggacaaa gcggacctac gactaatata      540

cctagcgcta gcgcacatga taaaattccg agggcacttc ctaatagaag gggacctaaa      600

cccggacaac agcgacgtag acaaactatt catacaacta gtacaaacgt acaaccaact      660

attcgaagaa aacccgataa acgcgagcgg ggtagacgcg aaagcgatac taagcgcgcg      720

actaagcaaa agccgacgac tagaaaacct aatagcgcaa ctaccggggg aaaaaaaaaa      780

cgggctattc gggaacctaa tagcgctaag cctagggcta acgccgaact tcaaaagcaa      840

cttcgaccta gcggaagacg cgaaactaca actaagcaaa gacacgtacg acgacgacct      900

agacaaccta ctagcgcaaa taggggacca atacgcggac ctattcctag cggcgaaaaa      960

cctaagcgac gcgatactac taagcgacat actacgagta aacacggaaa taacgaaagc     1020

gccgctaagc gcgagcatga taaaacgata cgacgaacac caccaagacc taacgctact     1080

aaaagcgcta gtacgacaac aactaccgga aaaatacaaa gaaatattct tcgaccaaag     1140

caaaaacggg tacgcggggt acatagacgg gggggcgagc caagaagaat tctacaaatt     1200

cataaaaccg atactagaaa aaatggacgg gacggaagaa ctactagtaa aactaaaccg     1260

agaagaccta ctacgaaaac aacgaacgtt cgacaacggg agcataccgc accaaataca     1320

cctaggggaa ctacacgcga tactacgacg acaagaagac ttctacccgt tcctaaaaga     1380

caaccgagaa aaaatagaaa aaatactaac gttccgaata ccgtactacg tagggccgct     1440

agcgcgaggg aacagccgat tcgcgtggat gacgcgaaaa agcgaagaaa cgataacgcc     1500

gtggaacttc gaagaagtag tagacaaagg ggcgagcgcg caaagcttca tagaacgaat     1560

gacgaacttc gacaaaaacc taccgaacga aaaagtacta ccgaaacaca gcctactata     1620

cgaatacttc acggtataca acgaactaac gaaagtaaaa tacgtaacgg aagggatgcg     1680

aaaaccggcg ttcctaagcg gggaacaaaa aaaagcgata gtagacctac tattcaaaac     1740

gaaccgaaaa gtaacggtaa aacaactaaa agaagactac ttcaaaaaaa tagaatgctt     1800

cgacagcgta gaaataagcg gggtagaaga ccgattcaac gcgagcctag ggacgtacca     1860

cgacctacta aaaataataa aagacaaaga cttcctagac aacgaagaaa acgaagacat     1920

actagaagac atagtactaa cgctaacgct attcgaagac cgagaaatga tagaagaacg     1980

actaaaaacg tacgcgcacc tattcgacga caaagtaatg aaacaactaa aacgacgacg     2040

atacacgggg tgggggcgac taagccgaaa actaataaac gggatacgag acaaacaaag     2100

cgggaaaacg atactagact tcctaaaaag cgacgggttc gcgaaccgaa acttcatgca     2160

actaatacac gacgacagcc taacgttcaa agaagacata caaaaagcgc aagtaagcgg     2220

gcaaggggac agcctacacg aacacatagc gaacctagcg gggagcccgg cgataaaaaa     2280

agggatacta caaacggtaa aagtagtaga cgaactagta aaagtaatgg ggcgacacaa     2340

accggaaaac atagtaatag aaatggcgcg agaaaaccaa acgacgcaaa aagggcaaaa     2400

aaacagccga gaacgaatga aacgaataga agaagggata aaagaactag ggagccaaat     2460

actaaaagaa cacccggtag aaaacacgca actacaaaac gaaaaactat acctatacta     2520

cctacaaaac gggcgagaca tgtacgtaga ccaagaacta gacataaacc gactaagcga     2580

ctacgacgta gaccacatag taccgcaaag cttcctaaaa gacgacagca tagacaacaa     2640

agtactaacg cgaagcgaca aaaaccgagg gaaaagcgac aacgtaccga gcgaagaagt     2700

agtaaaaaaa atgaaaaact actggcgaca actactaaac gcgaaactaa taacgcaacg     2760

aaaattcgac aacctaacga aagcggaacg aggggggcta agcgaactag acaaagcggg     2820

gttcataaaa cgacaactag tagaaacgcg acaaataacg aaacacgtag cgcaaatact     2880

agacagccga atgaacacga aatacgacga aaacgacaaa ctaatacgag aagtaaaagt     2940

aataacgcta aaaagcaaac tagtaagcga cttccgaaaa gacttccaat tctacaaagt     3000

acgagaaata aacaactacc accacgcgca cgacgcgtac ctaaacgcgg tagtagggac     3060

ggcgctaata aaaaaatacc cgaaactaga aagcgaattc gtatacgggg actacaaagt     3120

atacgacgta cgaaaaatga tagcgaaaag cgaacaagaa atagggaaag cgacggcgaa     3180

atacttcttc tacagcaaca taatgaactt cttcaaaacg gaaataacgc tagcgaacgg     3240

ggaaatacga aaacgaccgc taatagaaac gaacggggaa acgggggaaa tagtatggga     3300

caaagggcga gacttcgcga cggtacgaaa agtactaagc atgccgcaag taaacatagt     3360

aaaaaaaacg gaagtacaaa cgggggggtt cagcaaagaa agcatactac cgaaacgaaa     3420

cagcgacaaa ctaatagcgc gaaaaaaaga ctgggacccg aaaaaatacg gggggttcga     3480

cagcccgacg gtagcgtaca gcgtactagt agtagcgaaa gtagaaaaag ggaaaagcaa     3540

aaaactaaaa agcgtaaaag aactactagg gataacgata atggaacgaa gcagcttcga     3600

aaaaaacccg atagacttcc tagaagcgaa agggtacaaa gaagtaaaaa aagacctaat     3660

aataaaacta ccgaaataca gcctattcga actagaaaac gggcgaaaac gaatgctagc     3720

gagcgcgggg gaactacaaa aagggaacga actagcgcta ccgagcaaat acgtaaactt     3780

cctataccta gcgagccact acgaaaaact aaaagggagc ccggaagaca acgaacaaaa     3840

acaactattc gtagaacaac acaaacacta cctagacgaa ataatagaac aaataagcga     3900

attcagcaaa cgagtaatac tagcggacgc gaacctagac aaagtactaa gcgcgtacaa     3960

caaacaccga gacaaaccga tacgagaaca agcggaaaac ataatacacc tattcacgct     4020

aacgaaccta ggggcgccgg cggcgttcaa atacttcgac acgacgatag accgaaaacg     4080

atacacgagc acgaaagaag tactagacgc gacgctaata caccaaagca taacggggct     4140

atacgaaacg cgaatagacc taagccaact agggggggac ggggggggga gcccgaaaaa     4200

aaaacgaaaa gtatgactag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  56
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with AGG as first three nucleotides 
       for use with CleanCapTM, 5 UTR of HSD, ORF corresponding to SEQ 
       ID NO: 4, Kozak sequence, and 3 UTR of ALB

<400>  56
aggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  57
<211>  4481
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR from CMV, ORF corresponding
       to SEQ ID NO: 4, Kozak sequence, and 3 UTR of ALB

<400>  57
gggcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac       60

cgatccagcc tccgcggccg ggaacggtgc attggaacgc ggattccccg tgccaagagt      120

gactcaccgt ccttgacacg gccaccatgg acaagaagta cagcatcgga ctggacatcg      180

gaacaaacag cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt      240

tcaaggtcct gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc      300

tgttcgacag cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat      360

acacaagaag aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa      420

aggtcgacga cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga      480

agcacgaaag acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt      540

acccgacaat ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga      600

gactgatcta cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag      660

gagacctgaa cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat      720

acaaccagct gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc      780

tgagcgcaag actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag      840

aaaagaagaa cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact      900

tcaagagcaa cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg      960

acgacgacct ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg     1020

cagcaaagaa cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa     1080

tcacaaaggc accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc     1140

tgacactgct gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct     1200

tcgaccagag caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat     1260

tctacaagtt catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca     1320

agctgaacag agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc     1380

accagatcca cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt     1440

tcctgaagga caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg     1500

tcggaccgct ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa     1560

caatcacacc gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca     1620

tcgaaagaat gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca     1680

gcctgctgta cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag     1740

aaggaatgag aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc     1800

tgttcaagac aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga     1860

tcgaatgctt cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg     1920

gaacatacca cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa     1980

acgaagacat cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga     2040

tcgaagaaag actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga     2100

agagaagaag atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag     2160

acaagcagag cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa     2220

acttcatgca gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac     2280

aggtcagcgg acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg     2340

caatcaagaa gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg     2400

gaagacacaa gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga     2460

agggacagaa gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg     2520

gaagccagat cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt     2580

acctgtacta cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca     2640

gactgagcga ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca     2700

tcgacaacaa ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga     2760

gcgaagaagt cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga     2820

tcacacagag aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg     2880

acaaggcagg attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg     2940

cacagatcct ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag     3000

aagtcaaggt catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt     3060

tctacaaggt cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag     3120

tcgtcggaac agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag     3180

actacaaggt ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg     3240

caacagcaaa gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac     3300

tggcaaacgg agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa     3360

tcgtctggga caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg     3420

tcaacatcgt caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc     3480

cgaagagaaa cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg     3540

gaggattcga cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg     3600

gaaagagcaa gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa     3660

gcagcttcga aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga     3720

aggacctgat catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga     3780

gaatgctggc aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt     3840

acgtcaactt cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca     3900

acgaacagaa gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac     3960

agatcagcga attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga     4020

gcgcatacaa caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc     4080

tgttcacact gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg     4140

acagaaagag atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca     4200

tcacaggact gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa     4260

gcccgaagaa gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc     4320

atgagaataa gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt     4380

tggtgtaaag ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt     4440

tctctgtgct tcaattaata aaaaatggaa agaacctcga g                         4481


<210>  58
<211>  4348
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR from HBB, ORF corresponding
       to SEQ ID NO: 4, Kozak sequence, and 3 UTR of HBB

<400>  58
gggacatttg cttctgacac aactgtgttc actagcaacc tcaaacagac accggatctg       60

ccaccatgga caagaagtac agcatcggac tggacatcgg aacaaacagc gtcggatggg      120

cagtcatcac agacgaatac aaggtcccga gcaagaagtt caaggtcctg ggaaacacag      180

acagacacag catcaagaag aacctgatcg gagcactgct gttcgacagc ggagaaacag      240

cagaagcaac aagactgaag agaacagcaa gaagaagata cacaagaaga aagaacagaa      300

tctgctacct gcaggaaatc ttcagcaacg aaatggcaaa ggtcgacgac agcttcttcc      360

acagactgga agaaagcttc ctggtcgaag aagacaagaa gcacgaaaga cacccgatct      420

tcggaaacat cgtcgacgaa gtcgcatacc acgaaaagta cccgacaatc taccacctga      480

gaaagaagct ggtcgacagc acagacaagg cagacctgag actgatctac ctggcactgg      540

cacacatgat caagttcaga ggacacttcc tgatcgaagg agacctgaac ccggacaaca      600

gcgacgtcga caagctgttc atccagctgg tccagacata caaccagctg ttcgaagaaa      660

acccgatcaa cgcaagcgga gtcgacgcaa aggcaatcct gagcgcaaga ctgagcaaga      720

gcagaagact ggaaaacctg atcgcacagc tgccgggaga aaagaagaac ggactgttcg      780

gaaacctgat cgcactgagc ctgggactga caccgaactt caagagcaac ttcgacctgg      840

cagaagacgc aaagctgcag ctgagcaagg acacatacga cgacgacctg gacaacctgc      900

tggcacagat cggagaccag tacgcagacc tgttcctggc agcaaagaac ctgagcgacg      960

caatcctgct gagcgacatc ctgagagtca acacagaaat cacaaaggca ccgctgagcg     1020

caagcatgat caagagatac gacgaacacc accaggacct gacactgctg aaggcactgg     1080

tcagacagca gctgccggaa aagtacaagg aaatcttctt cgaccagagc aagaacggat     1140

acgcaggata catcgacgga ggagcaagcc aggaagaatt ctacaagttc atcaagccga     1200

tcctggaaaa gatggacgga acagaagaac tgctggtcaa gctgaacaga gaagacctgc     1260

tgagaaagca gagaacattc gacaacggaa gcatcccgca ccagatccac ctgggagaac     1320

tgcacgcaat cctgagaaga caggaagact tctacccgtt cctgaaggac aacagagaaa     1380

agatcgaaaa gatcctgaca ttcagaatcc cgtactacgt cggaccgctg gcaagaggaa     1440

acagcagatt cgcatggatg acaagaaaga gcgaagaaac aatcacaccg tggaacttcg     1500

aagaagtcgt cgacaaggga gcaagcgcac agagcttcat cgaaagaatg acaaacttcg     1560

acaagaacct gccgaacgaa aaggtcctgc cgaagcacag cctgctgtac gaatacttca     1620

cagtctacaa cgaactgaca aaggtcaagt acgtcacaga aggaatgaga aagccggcat     1680

tcctgagcgg agaacagaag aaggcaatcg tcgacctgct gttcaagaca aacagaaagg     1740

tcacagtcaa gcagctgaag gaagactact tcaagaagat cgaatgcttc gacagcgtcg     1800

aaatcagcgg agtcgaagac agattcaacg caagcctggg aacataccac gacctgctga     1860

agatcatcaa ggacaaggac ttcctggaca acgaagaaaa cgaagacatc ctggaagaca     1920

tcgtcctgac actgacactg ttcgaagaca gagaaatgat cgaagaaaga ctgaagacat     1980

acgcacacct gttcgacgac aaggtcatga agcagctgaa gagaagaaga tacacaggat     2040

ggggaagact gagcagaaag ctgatcaacg gaatcagaga caagcagagc ggaaagacaa     2100

tcctggactt cctgaagagc gacggattcg caaacagaaa cttcatgcag ctgatccacg     2160

acgacagcct gacattcaag gaagacatcc agaaggcaca ggtcagcgga cagggagaca     2220

gcctgcacga acacatcgca aacctggcag gaagcccggc aatcaagaag ggaatcctgc     2280

agacagtcaa ggtcgtcgac gaactggtca aggtcatggg aagacacaag ccggaaaaca     2340

tcgtcatcga aatggcaaga gaaaaccaga caacacagaa gggacagaag aacagcagag     2400

aaagaatgaa gagaatcgaa gaaggaatca aggaactggg aagccagatc ctgaaggaac     2460

acccggtcga aaacacacag ctgcagaacg aaaagctgta cctgtactac ctgcagaacg     2520

gaagagacat gtacgtcgac caggaactgg acatcaacag actgagcgac tacgacgtcg     2580

accacatcgt cccgcagagc ttcctgaagg acgacagcat cgacaacaag gtcctgacaa     2640

gaagcgacaa gaacagagga aagagcgaca acgtcccgag cgaagaagtc gtcaagaaga     2700

tgaagaacta ctggagacag ctgctgaacg caaagctgat cacacagaga aagttcgaca     2760

acctgacaaa ggcagagaga ggaggactga gcgaactgga caaggcagga ttcatcaaga     2820

gacagctggt cgaaacaaga cagatcacaa agcacgtcgc acagatcctg gacagcagaa     2880

tgaacacaaa gtacgacgaa aacgacaagc tgatcagaga agtcaaggtc atcacactga     2940

agagcaagct ggtcagcgac ttcagaaagg acttccagtt ctacaaggtc agagaaatca     3000

acaactacca ccacgcacac gacgcatacc tgaacgcagt cgtcggaaca gcactgatca     3060

agaagtaccc gaagctggaa agcgaattcg tctacggaga ctacaaggtc tacgacgtca     3120

gaaagatgat cgcaaagagc gaacaggaaa tcggaaaggc aacagcaaag tacttcttct     3180

acagcaacat catgaacttc ttcaagacag aaatcacact ggcaaacgga gaaatcagaa     3240

agagaccgct gatcgaaaca aacggagaaa caggagaaat cgtctgggac aagggaagag     3300

acttcgcaac agtcagaaag gtcctgagca tgccgcaggt caacatcgtc aagaagacag     3360

aagtccagac aggaggattc agcaaggaaa gcatcctgcc gaagagaaac agcgacaagc     3420

tgatcgcaag aaagaaggac tgggacccga agaagtacgg aggattcgac agcccgacag     3480

tcgcatacag cgtcctggtc gtcgcaaagg tcgaaaaggg aaagagcaag aagctgaaga     3540

gcgtcaagga actgctggga atcacaatca tggaaagaag cagcttcgaa aagaacccga     3600

tcgacttcct ggaagcaaag ggatacaagg aagtcaagaa ggacctgatc atcaagctgc     3660

cgaagtacag cctgttcgaa ctggaaaacg gaagaaagag aatgctggca agcgcaggag     3720

aactgcagaa gggaaacgaa ctggcactgc cgagcaagta cgtcaacttc ctgtacctgg     3780

caagccacta cgaaaagctg aagggaagcc cggaagacaa cgaacagaag cagctgttcg     3840

tcgaacagca caagcactac ctggacgaaa tcatcgaaca gatcagcgaa ttcagcaaga     3900

gagtcatcct ggcagacgca aacctggaca aggtcctgag cgcatacaac aagcacagag     3960

acaagccgat cagagaacag gcagaaaaca tcatccacct gttcacactg acaaacctgg     4020

gagcaccggc agcattcaag tacttcgaca caacaatcga cagaaagaga tacacaagca     4080

caaaggaagt cctggacgca acactgatcc accagagcat cacaggactg tacgaaacaa     4140

gaatcgacct gagccagctg ggaggagacg gaggaggaag cccgaagaag aagagaaagg     4200

tctagctagc gctcgctttc ttgctgtcca atttctatta aaggttcctt tgttccctaa     4260

gtccaactac taaactgggg gatattatga agggccttga gcatctggat tctgcctaat     4320

aaaaaacatt tattttcatt gcctcgag                                        4348


<210>  59
<211>  4325
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with 5 UTR from XBG, ORF corresponding
       to SEQ ID NO: 4, Kozak sequence, and 3 UTR of XBG

<400>  59
gggaagctca gaataaacgc tcaactttgg ccggatctgc caccatggac aagaagtaca       60

gcatcggact ggacatcgga acaaacagcg tcggatgggc agtcatcaca gacgaataca      120

aggtcccgag caagaagttc aaggtcctgg gaaacacaga cagacacagc atcaagaaga      180

acctgatcgg agcactgctg ttcgacagcg gagaaacagc agaagcaaca agactgaaga      240

gaacagcaag aagaagatac acaagaagaa agaacagaat ctgctacctg caggaaatct      300

tcagcaacga aatggcaaag gtcgacgaca gcttcttcca cagactggaa gaaagcttcc      360

tggtcgaaga agacaagaag cacgaaagac acccgatctt cggaaacatc gtcgacgaag      420

tcgcatacca cgaaaagtac ccgacaatct accacctgag aaagaagctg gtcgacagca      480

cagacaaggc agacctgaga ctgatctacc tggcactggc acacatgatc aagttcagag      540

gacacttcct gatcgaagga gacctgaacc cggacaacag cgacgtcgac aagctgttca      600

tccagctggt ccagacatac aaccagctgt tcgaagaaaa cccgatcaac gcaagcggag      660

tcgacgcaaa ggcaatcctg agcgcaagac tgagcaagag cagaagactg gaaaacctga      720

tcgcacagct gccgggagaa aagaagaacg gactgttcgg aaacctgatc gcactgagcc      780

tgggactgac accgaacttc aagagcaact tcgacctggc agaagacgca aagctgcagc      840

tgagcaagga cacatacgac gacgacctgg acaacctgct ggcacagatc ggagaccagt      900

acgcagacct gttcctggca gcaaagaacc tgagcgacgc aatcctgctg agcgacatcc      960

tgagagtcaa cacagaaatc acaaaggcac cgctgagcgc aagcatgatc aagagatacg     1020

acgaacacca ccaggacctg acactgctga aggcactggt cagacagcag ctgccggaaa     1080

agtacaagga aatcttcttc gaccagagca agaacggata cgcaggatac atcgacggag     1140

gagcaagcca ggaagaattc tacaagttca tcaagccgat cctggaaaag atggacggaa     1200

cagaagaact gctggtcaag ctgaacagag aagacctgct gagaaagcag agaacattcg     1260

acaacggaag catcccgcac cagatccacc tgggagaact gcacgcaatc ctgagaagac     1320

aggaagactt ctacccgttc ctgaaggaca acagagaaaa gatcgaaaag atcctgacat     1380

tcagaatccc gtactacgtc ggaccgctgg caagaggaaa cagcagattc gcatggatga     1440

caagaaagag cgaagaaaca atcacaccgt ggaacttcga agaagtcgtc gacaagggag     1500

caagcgcaca gagcttcatc gaaagaatga caaacttcga caagaacctg ccgaacgaaa     1560

aggtcctgcc gaagcacagc ctgctgtacg aatacttcac agtctacaac gaactgacaa     1620

aggtcaagta cgtcacagaa ggaatgagaa agccggcatt cctgagcgga gaacagaaga     1680

aggcaatcgt cgacctgctg ttcaagacaa acagaaaggt cacagtcaag cagctgaagg     1740

aagactactt caagaagatc gaatgcttcg acagcgtcga aatcagcgga gtcgaagaca     1800

gattcaacgc aagcctggga acataccacg acctgctgaa gatcatcaag gacaaggact     1860

tcctggacaa cgaagaaaac gaagacatcc tggaagacat cgtcctgaca ctgacactgt     1920

tcgaagacag agaaatgatc gaagaaagac tgaagacata cgcacacctg ttcgacgaca     1980

aggtcatgaa gcagctgaag agaagaagat acacaggatg gggaagactg agcagaaagc     2040

tgatcaacgg aatcagagac aagcagagcg gaaagacaat cctggacttc ctgaagagcg     2100

acggattcgc aaacagaaac ttcatgcagc tgatccacga cgacagcctg acattcaagg     2160

aagacatcca gaaggcacag gtcagcggac agggagacag cctgcacgaa cacatcgcaa     2220

acctggcagg aagcccggca atcaagaagg gaatcctgca gacagtcaag gtcgtcgacg     2280

aactggtcaa ggtcatggga agacacaagc cggaaaacat cgtcatcgaa atggcaagag     2340

aaaaccagac aacacagaag ggacagaaga acagcagaga aagaatgaag agaatcgaag     2400

aaggaatcaa ggaactggga agccagatcc tgaaggaaca cccggtcgaa aacacacagc     2460

tgcagaacga aaagctgtac ctgtactacc tgcagaacgg aagagacatg tacgtcgacc     2520

aggaactgga catcaacaga ctgagcgact acgacgtcga ccacatcgtc ccgcagagct     2580

tcctgaagga cgacagcatc gacaacaagg tcctgacaag aagcgacaag aacagaggaa     2640

agagcgacaa cgtcccgagc gaagaagtcg tcaagaagat gaagaactac tggagacagc     2700

tgctgaacgc aaagctgatc acacagagaa agttcgacaa cctgacaaag gcagagagag     2760

gaggactgag cgaactggac aaggcaggat tcatcaagag acagctggtc gaaacaagac     2820

agatcacaaa gcacgtcgca cagatcctgg acagcagaat gaacacaaag tacgacgaaa     2880

acgacaagct gatcagagaa gtcaaggtca tcacactgaa gagcaagctg gtcagcgact     2940

tcagaaagga cttccagttc tacaaggtca gagaaatcaa caactaccac cacgcacacg     3000

acgcatacct gaacgcagtc gtcggaacag cactgatcaa gaagtacccg aagctggaaa     3060

gcgaattcgt ctacggagac tacaaggtct acgacgtcag aaagatgatc gcaaagagcg     3120

aacaggaaat cggaaaggca acagcaaagt acttcttcta cagcaacatc atgaacttct     3180

tcaagacaga aatcacactg gcaaacggag aaatcagaaa gagaccgctg atcgaaacaa     3240

acggagaaac aggagaaatc gtctgggaca agggaagaga cttcgcaaca gtcagaaagg     3300

tcctgagcat gccgcaggtc aacatcgtca agaagacaga agtccagaca ggaggattca     3360

gcaaggaaag catcctgccg aagagaaaca gcgacaagct gatcgcaaga aagaaggact     3420

gggacccgaa gaagtacgga ggattcgaca gcccgacagt cgcatacagc gtcctggtcg     3480

tcgcaaaggt cgaaaaggga aagagcaaga agctgaagag cgtcaaggaa ctgctgggaa     3540

tcacaatcat ggaaagaagc agcttcgaaa agaacccgat cgacttcctg gaagcaaagg     3600

gatacaagga agtcaagaag gacctgatca tcaagctgcc gaagtacagc ctgttcgaac     3660

tggaaaacgg aagaaagaga atgctggcaa gcgcaggaga actgcagaag ggaaacgaac     3720

tggcactgcc gagcaagtac gtcaacttcc tgtacctggc aagccactac gaaaagctga     3780

agggaagccc ggaagacaac gaacagaagc agctgttcgt cgaacagcac aagcactacc     3840

tggacgaaat catcgaacag atcagcgaat tcagcaagag agtcatcctg gcagacgcaa     3900

acctggacaa ggtcctgagc gcatacaaca agcacagaga caagccgatc agagaacagg     3960

cagaaaacat catccacctg ttcacactga caaacctggg agcaccggca gcattcaagt     4020

acttcgacac aacaatcgac agaaagagat acacaagcac aaaggaagtc ctggacgcaa     4080

cactgatcca ccagagcatc acaggactgt acgaaacaag aatcgacctg agccagctgg     4140

gaggagacgg aggaggaagc ccgaagaaga agagaaaggt ctagctagca ccagcctcaa     4200

gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt     4260

cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattctc     4320

tcgag                                                                 4325


<210>  60
<211>  4325
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with AGG as first three nucleotides 
       for use with CleanCapTM, 5 UTR from XBG, ORF corresponding to SEQ
       ID NO: 4, Kozak sequence, and 3 UTR of XBG

<400>  60
aggaagctca gaataaacgc tcaactttgg ccggatctgc caccatggac aagaagtaca       60

gcatcggact ggacatcgga acaaacagcg tcggatgggc agtcatcaca gacgaataca      120

aggtcccgag caagaagttc aaggtcctgg gaaacacaga cagacacagc atcaagaaga      180

acctgatcgg agcactgctg ttcgacagcg gagaaacagc agaagcaaca agactgaaga      240

gaacagcaag aagaagatac acaagaagaa agaacagaat ctgctacctg caggaaatct      300

tcagcaacga aatggcaaag gtcgacgaca gcttcttcca cagactggaa gaaagcttcc      360

tggtcgaaga agacaagaag cacgaaagac acccgatctt cggaaacatc gtcgacgaag      420

tcgcatacca cgaaaagtac ccgacaatct accacctgag aaagaagctg gtcgacagca      480

cagacaaggc agacctgaga ctgatctacc tggcactggc acacatgatc aagttcagag      540

gacacttcct gatcgaagga gacctgaacc cggacaacag cgacgtcgac aagctgttca      600

tccagctggt ccagacatac aaccagctgt tcgaagaaaa cccgatcaac gcaagcggag      660

tcgacgcaaa ggcaatcctg agcgcaagac tgagcaagag cagaagactg gaaaacctga      720

tcgcacagct gccgggagaa aagaagaacg gactgttcgg aaacctgatc gcactgagcc      780

tgggactgac accgaacttc aagagcaact tcgacctggc agaagacgca aagctgcagc      840

tgagcaagga cacatacgac gacgacctgg acaacctgct ggcacagatc ggagaccagt      900

acgcagacct gttcctggca gcaaagaacc tgagcgacgc aatcctgctg agcgacatcc      960

tgagagtcaa cacagaaatc acaaaggcac cgctgagcgc aagcatgatc aagagatacg     1020

acgaacacca ccaggacctg acactgctga aggcactggt cagacagcag ctgccggaaa     1080

agtacaagga aatcttcttc gaccagagca agaacggata cgcaggatac atcgacggag     1140

gagcaagcca ggaagaattc tacaagttca tcaagccgat cctggaaaag atggacggaa     1200

cagaagaact gctggtcaag ctgaacagag aagacctgct gagaaagcag agaacattcg     1260

acaacggaag catcccgcac cagatccacc tgggagaact gcacgcaatc ctgagaagac     1320

aggaagactt ctacccgttc ctgaaggaca acagagaaaa gatcgaaaag atcctgacat     1380

tcagaatccc gtactacgtc ggaccgctgg caagaggaaa cagcagattc gcatggatga     1440

caagaaagag cgaagaaaca atcacaccgt ggaacttcga agaagtcgtc gacaagggag     1500

caagcgcaca gagcttcatc gaaagaatga caaacttcga caagaacctg ccgaacgaaa     1560

aggtcctgcc gaagcacagc ctgctgtacg aatacttcac agtctacaac gaactgacaa     1620

aggtcaagta cgtcacagaa ggaatgagaa agccggcatt cctgagcgga gaacagaaga     1680

aggcaatcgt cgacctgctg ttcaagacaa acagaaaggt cacagtcaag cagctgaagg     1740

aagactactt caagaagatc gaatgcttcg acagcgtcga aatcagcgga gtcgaagaca     1800

gattcaacgc aagcctggga acataccacg acctgctgaa gatcatcaag gacaaggact     1860

tcctggacaa cgaagaaaac gaagacatcc tggaagacat cgtcctgaca ctgacactgt     1920

tcgaagacag agaaatgatc gaagaaagac tgaagacata cgcacacctg ttcgacgaca     1980

aggtcatgaa gcagctgaag agaagaagat acacaggatg gggaagactg agcagaaagc     2040

tgatcaacgg aatcagagac aagcagagcg gaaagacaat cctggacttc ctgaagagcg     2100

acggattcgc aaacagaaac ttcatgcagc tgatccacga cgacagcctg acattcaagg     2160

aagacatcca gaaggcacag gtcagcggac agggagacag cctgcacgaa cacatcgcaa     2220

acctggcagg aagcccggca atcaagaagg gaatcctgca gacagtcaag gtcgtcgacg     2280

aactggtcaa ggtcatggga agacacaagc cggaaaacat cgtcatcgaa atggcaagag     2340

aaaaccagac aacacagaag ggacagaaga acagcagaga aagaatgaag agaatcgaag     2400

aaggaatcaa ggaactggga agccagatcc tgaaggaaca cccggtcgaa aacacacagc     2460

tgcagaacga aaagctgtac ctgtactacc tgcagaacgg aagagacatg tacgtcgacc     2520

aggaactgga catcaacaga ctgagcgact acgacgtcga ccacatcgtc ccgcagagct     2580

tcctgaagga cgacagcatc gacaacaagg tcctgacaag aagcgacaag aacagaggaa     2640

agagcgacaa cgtcccgagc gaagaagtcg tcaagaagat gaagaactac tggagacagc     2700

tgctgaacgc aaagctgatc acacagagaa agttcgacaa cctgacaaag gcagagagag     2760

gaggactgag cgaactggac aaggcaggat tcatcaagag acagctggtc gaaacaagac     2820

agatcacaaa gcacgtcgca cagatcctgg acagcagaat gaacacaaag tacgacgaaa     2880

acgacaagct gatcagagaa gtcaaggtca tcacactgaa gagcaagctg gtcagcgact     2940

tcagaaagga cttccagttc tacaaggtca gagaaatcaa caactaccac cacgcacacg     3000

acgcatacct gaacgcagtc gtcggaacag cactgatcaa gaagtacccg aagctggaaa     3060

gcgaattcgt ctacggagac tacaaggtct acgacgtcag aaagatgatc gcaaagagcg     3120

aacaggaaat cggaaaggca acagcaaagt acttcttcta cagcaacatc atgaacttct     3180

tcaagacaga aatcacactg gcaaacggag aaatcagaaa gagaccgctg atcgaaacaa     3240

acggagaaac aggagaaatc gtctgggaca agggaagaga cttcgcaaca gtcagaaagg     3300

tcctgagcat gccgcaggtc aacatcgtca agaagacaga agtccagaca ggaggattca     3360

gcaaggaaag catcctgccg aagagaaaca gcgacaagct gatcgcaaga aagaaggact     3420

gggacccgaa gaagtacgga ggattcgaca gcccgacagt cgcatacagc gtcctggtcg     3480

tcgcaaaggt cgaaaaggga aagagcaaga agctgaagag cgtcaaggaa ctgctgggaa     3540

tcacaatcat ggaaagaagc agcttcgaaa agaacccgat cgacttcctg gaagcaaagg     3600

gatacaagga agtcaagaag gacctgatca tcaagctgcc gaagtacagc ctgttcgaac     3660

tggaaaacgg aagaaagaga atgctggcaa gcgcaggaga actgcagaag ggaaacgaac     3720

tggcactgcc gagcaagtac gtcaacttcc tgtacctggc aagccactac gaaaagctga     3780

agggaagccc ggaagacaac gaacagaagc agctgttcgt cgaacagcac aagcactacc     3840

tggacgaaat catcgaacag atcagcgaat tcagcaagag agtcatcctg gcagacgcaa     3900

acctggacaa ggtcctgagc gcatacaaca agcacagaga caagccgatc agagaacagg     3960

cagaaaacat catccacctg ttcacactga caaacctggg agcaccggca gcattcaagt     4020

acttcgacac aacaatcgac agaaagagat acacaagcac aaaggaagtc ctggacgcaa     4080

cactgatcca ccagagcatc acaggactgt acgaaacaag aatcgacctg agccagctgg     4140

gaggagacgg aggaggaagc ccgaagaaga agagaaaggt ctagctagca ccagcctcaa     4200

gaacacccga atggagtctc taagctacat aataccaact tacactttac aaaatgttgt     4260

cccccaaaat gtagccattc gtatctgctc ctaataaaaa gaaagtttct tcacattctc     4320

tcgag                                                                 4325


<210>  61
<211>  4411
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 transcript with AGG as first three nucleotides 
       for use with CleanCapTM,  5 UTR from HSD, ORF corresponding to 
       SEQ ID NO: 4, Kozak sequence, and 3 UTR of ALB

<400>  61
aggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtctagctag ccatcacatt taaaagcatc tcagcctacc atgagaataa     4260

gagaaagaaa atgaagatca atagcttatt catctctttt tctttttcgt tggtgtaaag     4320

ccaacaccct gtctaaaaaa cataaatttc tttaatcatt ttgcctcttt tctctgtgct     4380

tcaattaata aaaaatggaa agaacctcga g                                    4411


<210>  62
<211>  105
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: 30/30/39 poly-A sequence

<400>  62
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa gcgaaaaaaa aaaaaaaaaa aaaaaaaaaa       60

aaaccgaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaa                      105


<210>  63
<211>  93
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: poly-A 100 sequence

<400>  63
aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa       60

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa                                    93


<210>  64
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G209 guide RNA


<220>
<221>  modified_base
<222>  (1)..(4)
<223>  PS linkage

<220>
<221>  modified_base
<222>  (1)..(3)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (96)..(99)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (96)..(100)
<223>  PS linkage

<400>  64
ccaguccagc gaggcaaagg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  65
<211>  3312
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: ORF encoding Neisseria meningitidis Cas9

<400>  65
atggcagcat tcaagccgaa ctcgatcaac tacatcctgg gactggacat cggaatcgca       60

tcggtcggat gggcaatggt cgaaatcgac gaagaagaaa acccgatcag actgatcgac      120

ctgggagtca gagtcttcga aagagcagaa gtcccgaaga caggagactc gctggcaatg      180

gcaagaagac tggcaagatc ggtcagaaga ctgacaagaa gaagagcaca cagactgctg      240

agaacaagaa gactgctgaa gagagaagga gtcctgcagg cagcaaactt cgacgaaaac      300

ggactgatca agtcgctgcc gaacacaccg tggcagctga gagcagcagc actggacaga      360

aagctgacac cgctggaatg gtcggcagtc ctgctgcacc tgatcaagca cagaggatac      420

ctgtcgcaga gaaagaacga aggagaaaca gcagacaagg aactgggagc actgctgaag      480

ggagtcgcag gaaacgcaca cgcactgcag acaggagact tcagaacacc ggcagaactg      540

gcactgaaca agttcgaaaa ggaatcggga cacatcagaa accagagatc ggactactcg      600

cacacattct cgagaaagga cctgcaggca gaactgatcc tgctgttcga aaagcagaag      660

gaattcggaa acccgcacgt ctcgggagga ctgaaggaag gaatcgaaac actgctgatg      720

acacagagac cggcactgtc gggagacgca gtccagaaga tgctgggaca ctgcacattc      780

gaaccggcag aaccgaaggc agcaaagaac acatacacag cagaaagatt catctggctg      840

acaaagctga acaacctgag aatcctggaa cagggatcgg aaagaccgct gacagacaca      900

gaaagagcaa cactgatgga cgaaccgtac agaaagtcga agctgacata cgcacaggca      960

agaaagctgc tgggactgga agacacagca ttcttcaagg gactgagata cggaaaggac     1020

aacgcagaag catcgacact gatggaaatg aaggcatacc acgcaatctc gagagcactg     1080

gaaaaggaag gactgaagga caagaagtcg ccgctgaacc tgtcgccgga actgcaggac     1140

gaaatcggaa cagcattctc gctgttcaag acagacgaag acatcacagg aagactgaag     1200

gacagaatcc agccggaaat cctggaagca ctgctgaagc acatctcgtt cgacaagttc     1260

gtccagatct cgctgaaggc actgagaaga atcgtcccgc tgatggaaca gggaaagaga     1320

tacgacgaag catgcgcaga aatctacgga gaccactacg gaaagaagaa cacagaagaa     1380

aagatctacc tgccgccgat cccggcagac gaaatcagaa acccggtcgt cctgagagca     1440

ctgtcgcagg caagaaaggt catcaacgga gtcgtcagaa gatacggatc gccggcaaga     1500

atccacatcg aaacagcaag agaagtcgga aagtcgttca aggacagaaa ggaaatcgaa     1560

aagagacagg aagaaaacag aaaggacaga gaaaaggcag cagcaaagtt cagagaatac     1620

ttcccgaact tcgtcggaga accgaagtcg aaggacatcc tgaagctgag actgtacgaa     1680

cagcagcacg gaaagtgcct gtactcggga aaggaaatca acctgggaag actgaacgaa     1740

aagggatacg tcgaaatcga ccacgcactg ccgttctcga gaacatggga cgactcgttc     1800

aacaacaagg tcctggtcct gggatcggaa aaccagaaca agggaaacca gacaccgtac     1860

gaatacttca acggaaagga caactcgaga gaatggcagg aattcaaggc aagagtcgaa     1920

acatcgagat tcccgagatc gaagaagcag agaatcctgc tgcagaagtt cgacgaagac     1980

ggattcaagg aaagaaacct gaacgacaca agatacgtca acagattcct gtgccagttc     2040

gtcgcagaca gaatgagact gacaggaaag ggaaagaaga gagtcttcgc atcgaacgga     2100

cagatcacaa acctgctgag aggattctgg ggactgagaa aggtcagagc agaaaacgac     2160

agacaccacg cactggacgc agtcgtcgtc gcatgctcga cagtcgcaat gcagcagaag     2220

atcacaagat tcgtcagata caaggaaatg aacgcattcg acggaaagac aatcgacaag     2280

gaaacaggag aagtcctgca ccagaagaca cacttcccgc agccgtggga attcttcgca     2340

caggaagtca tgatcagagt cttcggaaag ccggacggaa agccggaatt cgaagaagca     2400

gacacactgg aaaagctgag aacactgctg gcagaaaagc tgtcgtcgag accggaagca     2460

gtccacgaat acgtcacacc gctgttcgtc tcgagagcac cgaacagaaa gatgtcggga     2520

cagggacaca tggaaacagt caagtcggca aagagactgg acgaaggagt ctcggtcctg     2580

agagtcccgc tgacacagct gaagctgaag gacctggaaa agatggtcaa cagagaaaga     2640

gaaccgaagc tgtacgaagc actgaaggca agactggaag cacacaagga cgacccggca     2700

aaggcattcg cagaaccgtt ctacaagtac gacaaggcag gaaacagaac acagcaggtc     2760

aaggcagtca gagtcgaaca ggtccagaag acaggagtct gggtcagaaa ccacaacgga     2820

atcgcagaca acgcaacaat ggtcagagta gacgtcttcg aaaagggaga caagtactac     2880

ctggtcccga tctactcgtg gcaggtcgca aagggaatcc tgccggacag agcagtcgtc     2940

cagggaaagg acgaagaaga ctggcagctg atcgacgact cgttcaactt caagttctcg     3000

ctgcacccga acgacctggt cgaagtcatc acaaagaagg caagaatgtt cggatacttc     3060

gcatcgtgcc acagaggaac aggaaacatc aacatcagaa tccacgacct ggaccacaag     3120

atcggaaaga acggaatcct ggaaggaatc ggagtcaaga cagcactgtc gttccagaag     3180

taccagatcg acgaactggg aaaggaaatc agaccgtgca gactgaagaa gagaccgccg     3240

gtcagatccg gaaagagaac agcagacgga tcggaattcg aatcgccgaa gaagaagaga     3300

aaggtcgaat ga                                                         3312


<210>  66
<211>  3306
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: ORF encoding Neisseria meningitidis Cas9 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  66
gcagcattca agccgaactc gatcaactac atcctgggac tggacatcgg aatcgcatcg       60

gtcggatggg caatggtcga aatcgacgaa gaagaaaacc cgatcagact gatcgacctg      120

ggagtcagag tcttcgaaag agcagaagtc ccgaagacag gagactcgct ggcaatggca      180

agaagactgg caagatcggt cagaagactg acaagaagaa gagcacacag actgctgaga      240

acaagaagac tgctgaagag agaaggagtc ctgcaggcag caaacttcga cgaaaacgga      300

ctgatcaagt cgctgccgaa cacaccgtgg cagctgagag cagcagcact ggacagaaag      360

ctgacaccgc tggaatggtc ggcagtcctg ctgcacctga tcaagcacag aggatacctg      420

tcgcagagaa agaacgaagg agaaacagca gacaaggaac tgggagcact gctgaaggga      480

gtcgcaggaa acgcacacgc actgcagaca ggagacttca gaacaccggc agaactggca      540

ctgaacaagt tcgaaaagga atcgggacac atcagaaacc agagatcgga ctactcgcac      600

acattctcga gaaaggacct gcaggcagaa ctgatcctgc tgttcgaaaa gcagaaggaa      660

ttcggaaacc cgcacgtctc gggaggactg aaggaaggaa tcgaaacact gctgatgaca      720

cagagaccgg cactgtcggg agacgcagtc cagaagatgc tgggacactg cacattcgaa      780

ccggcagaac cgaaggcagc aaagaacaca tacacagcag aaagattcat ctggctgaca      840

aagctgaaca acctgagaat cctggaacag ggatcggaaa gaccgctgac agacacagaa      900

agagcaacac tgatggacga accgtacaga aagtcgaagc tgacatacgc acaggcaaga      960

aagctgctgg gactggaaga cacagcattc ttcaagggac tgagatacgg aaaggacaac     1020

gcagaagcat cgacactgat ggaaatgaag gcataccacg caatctcgag agcactggaa     1080

aaggaaggac tgaaggacaa gaagtcgccg ctgaacctgt cgccggaact gcaggacgaa     1140

atcggaacag cattctcgct gttcaagaca gacgaagaca tcacaggaag actgaaggac     1200

agaatccagc cggaaatcct ggaagcactg ctgaagcaca tctcgttcga caagttcgtc     1260

cagatctcgc tgaaggcact gagaagaatc gtcccgctga tggaacaggg aaagagatac     1320

gacgaagcat gcgcagaaat ctacggagac cactacggaa agaagaacac agaagaaaag     1380

atctacctgc cgccgatccc ggcagacgaa atcagaaacc cggtcgtcct gagagcactg     1440

tcgcaggcaa gaaaggtcat caacggagtc gtcagaagat acggatcgcc ggcaagaatc     1500

cacatcgaaa cagcaagaga agtcggaaag tcgttcaagg acagaaagga aatcgaaaag     1560

agacaggaag aaaacagaaa ggacagagaa aaggcagcag caaagttcag agaatacttc     1620

ccgaacttcg tcggagaacc gaagtcgaag gacatcctga agctgagact gtacgaacag     1680

cagcacggaa agtgcctgta ctcgggaaag gaaatcaacc tgggaagact gaacgaaaag     1740

ggatacgtcg aaatcgacca cgcactgccg ttctcgagaa catgggacga ctcgttcaac     1800

aacaaggtcc tggtcctggg atcggaaaac cagaacaagg gaaaccagac accgtacgaa     1860

tacttcaacg gaaaggacaa ctcgagagaa tggcaggaat tcaaggcaag agtcgaaaca     1920

tcgagattcc cgagatcgaa gaagcagaga atcctgctgc agaagttcga cgaagacgga     1980

ttcaaggaaa gaaacctgaa cgacacaaga tacgtcaaca gattcctgtg ccagttcgtc     2040

gcagacagaa tgagactgac aggaaaggga aagaagagag tcttcgcatc gaacggacag     2100

atcacaaacc tgctgagagg attctgggga ctgagaaagg tcagagcaga aaacgacaga     2160

caccacgcac tggacgcagt cgtcgtcgca tgctcgacag tcgcaatgca gcagaagatc     2220

acaagattcg tcagatacaa ggaaatgaac gcattcgacg gaaagacaat cgacaaggaa     2280

acaggagaag tcctgcacca gaagacacac ttcccgcagc cgtgggaatt cttcgcacag     2340

gaagtcatga tcagagtctt cggaaagccg gacggaaagc cggaattcga agaagcagac     2400

acactggaaa agctgagaac actgctggca gaaaagctgt cgtcgagacc ggaagcagtc     2460

cacgaatacg tcacaccgct gttcgtctcg agagcaccga acagaaagat gtcgggacag     2520

ggacacatgg aaacagtcaa gtcggcaaag agactggacg aaggagtctc ggtcctgaga     2580

gtcccgctga cacagctgaa gctgaaggac ctggaaaaga tggtcaacag agaaagagaa     2640

ccgaagctgt acgaagcact gaaggcaaga ctggaagcac acaaggacga cccggcaaag     2700

gcattcgcag aaccgttcta caagtacgac aaggcaggaa acagaacaca gcaggtcaag     2760

gcagtcagag tcgaacaggt ccagaagaca ggagtctggg tcagaaacca caacggaatc     2820

gcagacaacg caacaatggt cagagtagac gtcttcgaaa agggagacaa gtactacctg     2880

gtcccgatct actcgtggca ggtcgcaaag ggaatcctgc cggacagagc agtcgtccag     2940

ggaaaggacg aagaagactg gcagctgatc gacgactcgt tcaacttcaa gttctcgctg     3000

cacccgaacg acctggtcga agtcatcaca aagaaggcaa gaatgttcgg atacttcgca     3060

tcgtgccaca gaggaacagg aaacatcaac atcagaatcc acgacctgga ccacaagatc     3120

ggaaagaacg gaatcctgga aggaatcgga gtcaagacag cactgtcgtt ccagaagtac     3180

cagatcgacg aactgggaaa ggaaatcaga ccgtgcagac tgaagaagag accgccggtc     3240

agatccggaa agagaacagc agacggatcg gaattcgaat cgccgaagaa gaagagaaag     3300

gtcgaa                                                                3306


<210>  67
<211>  3636
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Transcript comprising SEQ ID NO: 65 (encoding 
       Neisseria meningitidis Cas9)

<400>  67
gggagaccca agctggctag cgtttaaact taagcttgga tccgccacca tggcagcatt       60

caagccgaac tcgatcaact acatcctggg actggacatc ggaatcgcat cggtcggatg      120

ggcaatggtc gaaatcgacg aagaagaaaa cccgatcaga ctgatcgacc tgggagtcag      180

agtcttcgaa agagcagaag tcccgaagac aggagactcg ctggcaatgg caagaagact      240

ggcaagatcg gtcagaagac tgacaagaag aagagcacac agactgctga gaacaagaag      300

actgctgaag agagaaggag tcctgcaggc agcaaacttc gacgaaaacg gactgatcaa      360

gtcgctgccg aacacaccgt ggcagctgag agcagcagca ctggacagaa agctgacacc      420

gctggaatgg tcggcagtcc tgctgcacct gatcaagcac agaggatacc tgtcgcagag      480

aaagaacgaa ggagaaacag cagacaagga actgggagca ctgctgaagg gagtcgcagg      540

aaacgcacac gcactgcaga caggagactt cagaacaccg gcagaactgg cactgaacaa      600

gttcgaaaag gaatcgggac acatcagaaa ccagagatcg gactactcgc acacattctc      660

gagaaaggac ctgcaggcag aactgatcct gctgttcgaa aagcagaagg aattcggaaa      720

cccgcacgtc tcgggaggac tgaaggaagg aatcgaaaca ctgctgatga cacagagacc      780

ggcactgtcg ggagacgcag tccagaagat gctgggacac tgcacattcg aaccggcaga      840

accgaaggca gcaaagaaca catacacagc agaaagattc atctggctga caaagctgaa      900

caacctgaga atcctggaac agggatcgga aagaccgctg acagacacag aaagagcaac      960

actgatggac gaaccgtaca gaaagtcgaa gctgacatac gcacaggcaa gaaagctgct     1020

gggactggaa gacacagcat tcttcaaggg actgagatac ggaaaggaca acgcagaagc     1080

atcgacactg atggaaatga aggcatacca cgcaatctcg agagcactgg aaaaggaagg     1140

actgaaggac aagaagtcgc cgctgaacct gtcgccggaa ctgcaggacg aaatcggaac     1200

agcattctcg ctgttcaaga cagacgaaga catcacagga agactgaagg acagaatcca     1260

gccggaaatc ctggaagcac tgctgaagca catctcgttc gacaagttcg tccagatctc     1320

gctgaaggca ctgagaagaa tcgtcccgct gatggaacag ggaaagagat acgacgaagc     1380

atgcgcagaa atctacggag accactacgg aaagaagaac acagaagaaa agatctacct     1440

gccgccgatc ccggcagacg aaatcagaaa cccggtcgtc ctgagagcac tgtcgcaggc     1500

aagaaaggtc atcaacggag tcgtcagaag atacggatcg ccggcaagaa tccacatcga     1560

aacagcaaga gaagtcggaa agtcgttcaa ggacagaaag gaaatcgaaa agagacagga     1620

agaaaacaga aaggacagag aaaaggcagc agcaaagttc agagaatact tcccgaactt     1680

cgtcggagaa ccgaagtcga aggacatcct gaagctgaga ctgtacgaac agcagcacgg     1740

aaagtgcctg tactcgggaa aggaaatcaa cctgggaaga ctgaacgaaa agggatacgt     1800

cgaaatcgac cacgcactgc cgttctcgag aacatgggac gactcgttca acaacaaggt     1860

cctggtcctg ggatcggaaa accagaacaa gggaaaccag acaccgtacg aatacttcaa     1920

cggaaaggac aactcgagag aatggcagga attcaaggca agagtcgaaa catcgagatt     1980

cccgagatcg aagaagcaga gaatcctgct gcagaagttc gacgaagacg gattcaagga     2040

aagaaacctg aacgacacaa gatacgtcaa cagattcctg tgccagttcg tcgcagacag     2100

aatgagactg acaggaaagg gaaagaagag agtcttcgca tcgaacggac agatcacaaa     2160

cctgctgaga ggattctggg gactgagaaa ggtcagagca gaaaacgaca gacaccacgc     2220

actggacgca gtcgtcgtcg catgctcgac agtcgcaatg cagcagaaga tcacaagatt     2280

cgtcagatac aaggaaatga acgcattcga cggaaagaca atcgacaagg aaacaggaga     2340

agtcctgcac cagaagacac acttcccgca gccgtgggaa ttcttcgcac aggaagtcat     2400

gatcagagtc ttcggaaagc cggacggaaa gccggaattc gaagaagcag acacactgga     2460

aaagctgaga acactgctgg cagaaaagct gtcgtcgaga ccggaagcag tccacgaata     2520

cgtcacaccg ctgttcgtct cgagagcacc gaacagaaag atgtcgggac agggacacat     2580

ggaaacagtc aagtcggcaa agagactgga cgaaggagtc tcggtcctga gagtcccgct     2640

gacacagctg aagctgaagg acctggaaaa gatggtcaac agagaaagag aaccgaagct     2700

gtacgaagca ctgaaggcaa gactggaagc acacaaggac gacccggcaa aggcattcgc     2760

agaaccgttc tacaagtacg acaaggcagg aaacagaaca cagcaggtca aggcagtcag     2820

agtcgaacag gtccagaaga caggagtctg ggtcagaaac cacaacggaa tcgcagacaa     2880

cgcaacaatg gtcagagtag acgtcttcga aaagggagac aagtactacc tggtcccgat     2940

ctactcgtgg caggtcgcaa agggaatcct gccggacaga gcagtcgtcc agggaaagga     3000

cgaagaagac tggcagctga tcgacgactc gttcaacttc aagttctcgc tgcacccgaa     3060

cgacctggtc gaagtcatca caaagaaggc aagaatgttc ggatacttcg catcgtgcca     3120

cagaggaaca ggaaacatca acatcagaat ccacgacctg gaccacaaga tcggaaagaa     3180

cggaatcctg gaaggaatcg gagtcaagac agcactgtcg ttccagaagt accagatcga     3240

cgaactggga aaggaaatca gaccgtgcag actgaagaag agaccgccgg tcagatccgg     3300

aaagagaaca gcagacggat cggaattcga atcgccgaag aagaagagaa aggtcgaatg     3360

atagctagct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt     3420

ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg     3480

ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt     3540

gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca     3600

atagcaggca tgctggggat gcggtgggct ctatgg                               3636


<210>  68
<211>  1103
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence of Neisseria meningitidis Cas9

<400>  68

Met Ala Ala Phe Lys Pro Asn Ser Ile Asn Tyr Ile Leu Gly Leu Asp 
1               5                   10                  15      


Ile Gly Ile Ala Ser Val Gly Trp Ala Met Val Glu Ile Asp Glu Glu 
            20                  25                  30          


Glu Asn Pro Ile Arg Leu Ile Asp Leu Gly Val Arg Val Phe Glu Arg 
        35                  40                  45              


Ala Glu Val Pro Lys Thr Gly Asp Ser Leu Ala Met Ala Arg Arg Leu 
    50                  55                  60                  


Ala Arg Ser Val Arg Arg Leu Thr Arg Arg Arg Ala His Arg Leu Leu 
65                  70                  75                  80  


Arg Thr Arg Arg Leu Leu Lys Arg Glu Gly Val Leu Gln Ala Ala Asn 
                85                  90                  95      


Phe Asp Glu Asn Gly Leu Ile Lys Ser Leu Pro Asn Thr Pro Trp Gln 
            100                 105                 110         


Leu Arg Ala Ala Ala Leu Asp Arg Lys Leu Thr Pro Leu Glu Trp Ser 
        115                 120                 125             


Ala Val Leu Leu His Leu Ile Lys His Arg Gly Tyr Leu Ser Gln Arg 
    130                 135                 140                 


Lys Asn Glu Gly Glu Thr Ala Asp Lys Glu Leu Gly Ala Leu Leu Lys 
145                 150                 155                 160 


Gly Val Ala Gly Asn Ala His Ala Leu Gln Thr Gly Asp Phe Arg Thr 
                165                 170                 175     


Pro Ala Glu Leu Ala Leu Asn Lys Phe Glu Lys Glu Ser Gly His Ile 
            180                 185                 190         


Arg Asn Gln Arg Ser Asp Tyr Ser His Thr Phe Ser Arg Lys Asp Leu 
        195                 200                 205             


Gln Ala Glu Leu Ile Leu Leu Phe Glu Lys Gln Lys Glu Phe Gly Asn 
    210                 215                 220                 


Pro His Val Ser Gly Gly Leu Lys Glu Gly Ile Glu Thr Leu Leu Met 
225                 230                 235                 240 


Thr Gln Arg Pro Ala Leu Ser Gly Asp Ala Val Gln Lys Met Leu Gly 
                245                 250                 255     


His Cys Thr Phe Glu Pro Ala Glu Pro Lys Ala Ala Lys Asn Thr Tyr 
            260                 265                 270         


Thr Ala Glu Arg Phe Ile Trp Leu Thr Lys Leu Asn Asn Leu Arg Ile 
        275                 280                 285             


Leu Glu Gln Gly Ser Glu Arg Pro Leu Thr Asp Thr Glu Arg Ala Thr 
    290                 295                 300                 


Leu Met Asp Glu Pro Tyr Arg Lys Ser Lys Leu Thr Tyr Ala Gln Ala 
305                 310                 315                 320 


Arg Lys Leu Leu Gly Leu Glu Asp Thr Ala Phe Phe Lys Gly Leu Arg 
                325                 330                 335     


Tyr Gly Lys Asp Asn Ala Glu Ala Ser Thr Leu Met Glu Met Lys Ala 
            340                 345                 350         


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


Lys Ser Pro Leu Asn Leu Ser Pro Glu Leu Gln Asp Glu Ile Gly Thr 
    370                 375                 380                 


Ala Phe Ser Leu Phe Lys Thr Asp Glu Asp Ile Thr Gly Arg Leu Lys 
385                 390                 395                 400 


Asp Arg Ile Gln Pro Glu Ile Leu Glu Ala Leu Leu Lys His Ile Ser 
                405                 410                 415     


Phe Asp Lys Phe Val Gln Ile Ser Leu Lys Ala Leu Arg Arg Ile Val 
            420                 425                 430         


Pro Leu Met Glu Gln Gly Lys Arg Tyr Asp Glu Ala Cys Ala Glu Ile 
        435                 440                 445             


Tyr Gly Asp His Tyr Gly Lys Lys Asn Thr Glu Glu Lys Ile Tyr Leu 
    450                 455                 460                 


Pro Pro Ile Pro Ala Asp Glu Ile Arg Asn Pro Val Val Leu Arg Ala 
465                 470                 475                 480 


Leu Ser Gln Ala Arg Lys Val Ile Asn Gly Val Val Arg Arg Tyr Gly 
                485                 490                 495     


Ser Pro Ala Arg Ile His Ile Glu Thr Ala Arg Glu Val Gly Lys Ser 
            500                 505                 510         


Phe Lys Asp Arg Lys Glu Ile Glu Lys Arg Gln Glu Glu Asn Arg Lys 
        515                 520                 525             


Asp Arg Glu Lys Ala Ala Ala Lys Phe Arg Glu Tyr Phe Pro Asn Phe 
    530                 535                 540                 


Val Gly Glu Pro Lys Ser Lys Asp Ile Leu Lys Leu Arg Leu Tyr Glu 
545                 550                 555                 560 


Gln Gln His Gly Lys Cys Leu Tyr Ser Gly Lys Glu Ile Asn Leu Gly 
                565                 570                 575     


Arg Leu Asn Glu Lys Gly Tyr Val Glu Ile Asp His Ala Leu Pro Phe 
            580                 585                 590         


Ser Arg Thr Trp Asp Asp Ser Phe Asn Asn Lys Val Leu Val Leu Gly 
        595                 600                 605             


Ser Glu Asn Gln Asn Lys Gly Asn Gln Thr Pro Tyr Glu Tyr Phe Asn 
    610                 615                 620                 


Gly Lys Asp Asn Ser Arg Glu Trp Gln Glu Phe Lys Ala Arg Val Glu 
625                 630                 635                 640 


Thr Ser Arg Phe Pro Arg Ser Lys Lys Gln Arg Ile Leu Leu Gln Lys 
                645                 650                 655     


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


Val Asn Arg Phe Leu Cys Gln Phe Val Ala Asp Arg Met Arg Leu Thr 
        675                 680                 685             


Gly Lys Gly Lys Lys Arg Val Phe Ala Ser Asn Gly Gln Ile Thr Asn 
    690                 695                 700                 


Leu Leu Arg Gly Phe Trp Gly Leu Arg Lys Val Arg Ala Glu Asn Asp 
705                 710                 715                 720 


Arg His His Ala Leu Asp Ala Val Val Val Ala Cys Ser Thr Val Ala 
                725                 730                 735     


Met Gln Gln Lys Ile Thr Arg Phe Val Arg Tyr Lys Glu Met Asn Ala 
            740                 745                 750         


Phe Asp Gly Lys Thr Ile Asp Lys Glu Thr Gly Glu Val Leu His Gln 
        755                 760                 765             


Lys Thr His Phe Pro Gln Pro Trp Glu Phe Phe Ala Gln Glu Val Met 
    770                 775                 780                 


Ile Arg Val Phe Gly Lys Pro Asp Gly Lys Pro Glu Phe Glu Glu Ala 
785                 790                 795                 800 


Asp Thr Leu Glu Lys Leu Arg Thr Leu Leu Ala Glu Lys Leu Ser Ser 
                805                 810                 815     


Arg Pro Glu Ala Val His Glu Tyr Val Thr Pro Leu Phe Val Ser Arg 
            820                 825                 830         


Ala Pro Asn Arg Lys Met Ser Gly Gln Gly His Met Glu Thr Val Lys 
        835                 840                 845             


Ser Ala Lys Arg Leu Asp Glu Gly Val Ser Val Leu Arg Val Pro Leu 
    850                 855                 860                 


Thr Gln Leu Lys Leu Lys Asp Leu Glu Lys Met Val Asn Arg Glu Arg 
865                 870                 875                 880 


Glu Pro Lys Leu Tyr Glu Ala Leu Lys Ala Arg Leu Glu Ala His Lys 
                885                 890                 895     


Asp Asp Pro Ala Lys Ala Phe Ala Glu Pro Phe Tyr Lys Tyr Asp Lys 
            900                 905                 910         


Ala Gly Asn Arg Thr Gln Gln Val Lys Ala Val Arg Val Glu Gln Val 
        915                 920                 925             


Gln Lys Thr Gly Val Trp Val Arg Asn His Asn Gly Ile Ala Asp Asn 
    930                 935                 940                 


Ala Thr Met Val Arg Val Asp Val Phe Glu Lys Gly Asp Lys Tyr Tyr 
945                 950                 955                 960 


Leu Val Pro Ile Tyr Ser Trp Gln Val Ala Lys Gly Ile Leu Pro Asp 
                965                 970                 975     


Arg Ala Val Val Gln Gly Lys Asp Glu Glu Asp Trp Gln Leu Ile Asp 
            980                 985                 990         


Asp Ser Phe Asn Phe Lys Phe Ser  Leu His Pro Asn Asp  Leu Val Glu 
        995                 1000                 1005             


Val Ile  Thr Lys Lys Ala Arg  Met Phe Gly Tyr Phe  Ala Ser Cys 
    1010                 1015                 1020             


His Arg  Gly Thr Gly Asn Ile  Asn Ile Arg Ile His  Asp Leu Asp 
    1025                 1030                 1035             


His Lys  Ile Gly Lys Asn Gly  Ile Leu Glu Gly Ile  Gly Val Lys 
    1040                 1045                 1050             


Thr Ala  Leu Ser Phe Gln Lys  Tyr Gln Ile Asp Glu  Leu Gly Lys 
    1055                 1060                 1065             


Glu Ile  Arg Pro Cys Arg Leu  Lys Lys Arg Pro Pro  Val Arg Ser 
    1070                 1075                 1080             


Gly Lys  Arg Thr Ala Asp Gly  Ser Glu Phe Glu Ser  Pro Lys Lys 
    1085                 1090                 1095             


Lys Arg  Lys Val Glu 
    1100             


<210>  69
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G390 guide RNA


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  69
gccgagucug gagagcugca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  70
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G502 guide RNA


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  70
acacaaauac caguccagcg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  71
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G509 guide RNA


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  71
aaaguucuag augccguccg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  72
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G534 guide RNA


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  72
acgcaaauau caguccagcg guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  73
<211>  3783
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: DNA coding sequence of eGFP

<400>  73
tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca       60

cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg      120

ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc      180

accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc      240

attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat      300

tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt      360

tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ctaatacgac tcactatagg      420

gtcccgcagt cggcgtccag cggctctgct tgttcgtgtg tgtgtcgttg caggccttat      480

tcggatccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg      540

agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg      600

ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct      660

ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc      720

acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca      780

ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg      840

acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc      900

tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc      960

agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc     1020

agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg     1080

acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc     1140

acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt     1200

acaagtaata ggaattatgc agtctagcca tcacatttaa aagcatctca gcctaccatg     1260

agaataagag aaagaaaatg aagatcaata gcttattcat ctctttttct ttttcgttgg     1320

tgtaaagcca acaccctgtc taaaaaacat aaatttcttt aatcattttg cctcttttct     1380

ctgtgcttca attaataaaa aatggaaaga acctcgagaa aaaaaaaaaa aaaaaaaaaa     1440

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     1500

aaaaaaaaaa aaaaaaaatc tagacttaag cttgatgagc tctagcttgg cgtaatcatg     1560

gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc     1620

cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc     1680

gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat     1740

cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac     1800

tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt     1860

aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca     1920

gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc     1980

ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact     2040

ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct     2100

gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag     2160

ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca     2220

cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa     2280

cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc     2340

gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag     2400

aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg     2460

tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca     2520

gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc     2580

tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag     2640

gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata     2700

tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat     2760

ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg     2820

ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc     2880

tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc     2940

aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc     3000

gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc     3060

gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc     3120

ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa     3180

gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat     3240

gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata     3300

gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca     3360

tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag     3420

gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc     3480

agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc     3540

aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata     3600

ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta     3660

gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta     3720

agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg     3780

tcg                                                                   3783


<210>  74
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Modified sgRNA pattern, where N are nucleotides 
       encoding a guide seqnence


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  74
nnnnnnnnnn nnnnnnnnnn guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  75
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CMV-1 5 UTR

<400>  75
cagatcgcct ggagacgcca tccacgctgt tttgacctcc at                          42


<210>  76
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CMV-2 5 UTR

<400>  76
agaagacacc gggaccgatc cagcctccgc ggccgggaac gg                          42


<210>  77
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: CMV-3 5 UTR

<400>  77
tgcattggaa cgcggattcc ccgtgccaag agtgactcac cg                          42


<210>  78
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: SV40 NLS

<400>  78

Pro Lys Lys Lys Arg Lys Val 
1               5           


<210>  79
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 1

<400>  79

Leu Ala Ala Lys Arg Ser Arg Thr Thr 
1               5                   


<210>  80
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 2

<400>  80

Gln Ala Ala Lys Arg Ser Arg Thr Thr 
1               5                   


<210>  81
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 3

<400>  81

Pro Ala Pro Ala Lys Arg Glu Arg Thr Thr 
1               5                   10  


<210>  82
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 4

<400>  82

Gln Ala Ala Lys Arg Pro Arg Thr Thr 
1               5                   


<210>  83
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 5

<400>  83

Arg Ala Ala Lys Arg Pro Arg Thr Thr 
1               5                   


<210>  84
<211>  11
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 6

<400>  84

Ala Ala Ala Lys Arg Ser Trp Ser Met Ala Ala 
1               5                   10      


<210>  85
<211>  11
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 7

<400>  85

Ala Ala Ala Lys Arg Val Trp Ser Met Ala Phe 
1               5                   10      


<210>  86
<211>  11
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 8

<400>  86

Ala Ala Ala Lys Arg Ser Trp Ser Met Ala Phe 
1               5                   10      


<210>  87
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 9

<400>  87

Ala Ala Ala Lys Arg Lys Tyr Phe Ala Ala 
1               5                   10  


<210>  88
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 10

<400>  88

Arg Ala Ala Lys Arg Lys Ala Phe Ala Ala 
1               5                   10  


<210>  89
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary NLS 11

<400>  89

Arg Ala Ala Lys Arg Lys Tyr Phe Ala Val 
1               5                   10  


<210>  90
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Alternate SV40 NLS

<400>  90

Pro Lys Lys Lys Arg Arg Val 
1               5           


<210>  91
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Nucleoplasmin NLS

<400>  91

Lys Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys 
1               5                   10                  15      


<210>  92
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for SV40 NLS

<400>  92
ccgaagaaga agagaaaggt c                                                 21


<210>  93
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS1

<400>  93
ctggcagcaa agagaagcag aacaaca                                           27


<210>  94
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS2

<400>  94
caggcagcaa agagaagcag aacaaca                                           27


<210>  95
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS3

<400>  95
ccggcaccgg caaagagaga aagaacaaca                                        30


<210>  96
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS4

<400>  96
caggcagcaa agagaccgag aacaaca                                           27


<210>  97
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS5

<400>  97
agagcagcaa agagaccgag aacaaca                                           27


<210>  98
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS6

<400>  98
gcagcagcaa agagaagctg gagcatggca gca                                    33


<210>  99
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS7

<400>  99
gcagcagcaa agagagtctg gagcatggca ttc                                    33


<210>  100
<211>  33
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS8

<400>  100
gcagcagcaa agagaagctg gagcatggca ttc                                    33


<210>  101
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS9

<400>  101
gcagcagcaa agagaaagta cttcgcagca                                        30


<210>  102
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS10

<400>  102
agagcagcaa agagaaaggc attcgcagca                                        30


<210>  103
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for NLS11

<400>  103
agagcagcaa agagaaagta cttcgcagtc                                        30


<210>  104
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Exemplary coding sequence for alternate SV40 NLS

<400>  104
ccgaagaaga agagaagagt c                                                 21


<210>  105
<211>  13
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: exemplary Kozak sequence

<400>  105
gccgccrcca ugg                                                          13


<210>  106
<211>  0
<212>  DNA
<213>  N/A

<400>  106
000

<210>  107
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using long half life codons of Table 4, with 
       start and stop codons

<400>  107
atggacaaga agtactctat cggtttggac atcggtacca actctgtcgg ttgggccgtc       60

atcaccgacg aatacaaggt cccatctaag aagttcaagg tcttgggtaa caccgacaga      120

cactctatca agaagaactt gatcggtgcc ttgttgttcg actctggtga aaccgccgaa      180

gccaccagat tgaagagaac cgccagaaga agatacacca gaagaaagaa cagaatctgc      240

tacttgcaag aaatcttctc taacgaaatg gccaaggtcg acgactcttt cttccacaga      300

ttggaagaat ctttcttggt cgaagaagac aagaagcacg aaagacaccc aatcttcggt      360

aacatcgtcg acgaagtcgc ctaccacgaa aagtacccaa ccatctacca cttgagaaag      420

aagttggtcg actctaccga caaggccgac ttgagattga tctacttggc cttggcccac      480

atgatcaagt tcagaggtca cttcttgatc gaaggtgact tgaacccaga caactctgac      540

gtcgacaagt tgttcatcca attggtccaa acctacaacc aattgttcga agaaaaccca      600

atcaacgcct ctggtgtcga cgccaaggcc atcttgtctg ccagattgtc taagagcaga      660

agattggaaa acttgatcgc ccaattgcca ggtgaaaaga agaacggttt gttcggtaac      720

ttgatcgcct tgtctttggg tttgacccca aacttcaagt ctaacttcga cttggccgaa      780

gacgccaagt tgcaattgtc taaggacacc tacgacgacg acttggacaa cttgttggcc      840

caaatcggtg accaatacgc cgacttgttc ttggccgcca agaacttgtc tgacgccatc      900

ttgttgtctg acatcttgag agtcaacacc gaaatcacca aggccccatt gtctgcctct      960

atgatcaaga gatacgacga acaccaccaa gacttgacct tgttgaaggc cttggtcaga     1020

caacaattgc cagaaaagta caaggaaatc ttcttcgacc aatctaagaa cggttacgcc     1080

ggttacatcg acggtggtgc ctctcaagaa gaattctaca agttcatcaa gccaatcttg     1140

gaaaagatgg acggtaccga agaattgttg gtcaagttga acagagaaga cttgttgaga     1200

aagcaaagaa ccttcgacaa cggttctatc ccacaccaaa tccacttggg tgaattgcac     1260

gccatcttga gaagacaaga agacttctac ccattcttga aggacaacag agaaaagatc     1320

gaaaagatct tgaccttcag aatcccatac tacgtcggtc cattggccag aggtaacagc     1380

agattcgcct ggatgaccag aaagtctgaa gaaaccatca ccccatggaa cttcgaagaa     1440

gtcgtcgaca agggtgcctc tgcccaatct ttcatcgaaa gaatgaccaa cttcgacaag     1500

aacttgccaa acgaaaaggt cttgccaaag cactctttgt tgtacgaata cttcaccgtc     1560

tacaacgaat tgaccaaggt caagtacgtc accgaaggta tgagaaagcc agccttcttg     1620

tctggtgaac aaaagaaggc catcgtcgac ttgttgttca agaccaacag aaaggtcacc     1680

gtcaagcaat tgaaggaaga ctacttcaag aagatcgaat gcttcgactc tgtcgaaatc     1740

tctggtgtcg aagacagatt caacgcctct ttgggtacct accacgactt gttgaagatc     1800

atcaaggaca aggacttctt ggacaacgaa gaaaacgaag acatcttgga agacatcgtc     1860

ttgaccttga ccttgttcga agacagagaa atgatcgaag aaagattgaa gacctacgcc     1920

cacttgttcg acgacaaggt catgaagcaa ttgaagagaa gaagatacac cggttggggt     1980

agattgagca gaaagttgat caacggtatc agagacaagc aatctggtaa gaccatcttg     2040

gacttcttga agtctgacgg tttcgccaac agaaacttca tgcaattgat ccacgacgac     2100

tctttgacct tcaaggaaga catccaaaag gcccaagtct ctggtcaagg tgactctttg     2160

cacgaacaca tcgccaactt ggccggttct ccagccatca agaagggtat cttgcaaacc     2220

gtcaaggtcg tcgacgaatt ggtcaaggtc atgggtagac acaagccaga aaacatcgtc     2280

atcgaaatgg ccagagaaaa ccaaaccacc caaaagggtc aaaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg tatcaaggaa ttgggttctc aaatcttgaa ggaacaccca     2400

gtcgaaaaca cccaattgca aaacgaaaag ttgtacttgt actacttgca aaacggtaga     2460

gacatgtacg tcgaccaaga attggacatc aacagattgt ctgactacga cgtcgaccac     2520

atcgtcccac aatctttctt gaaggacgac tctatcgaca acaaggtctt gaccagatct     2580

gacaagaaca gaggtaagtc tgacaacgtc ccatctgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacaattgtt gaacgccaag ttgatcaccc aaagaaagtt cgacaacttg     2700

accaaggccg aaagaggtgg tttgtctgaa ttggacaagg ccggtttcat caagagacaa     2760

ttggtcgaaa ccagacaaat caccaagcac gtcgcccaaa tcttggacag cagaatgaac     2820

accaagtacg acgaaaacga caagttgatc agagaagtca aggtcatcac cttgaagtct     2880

aagttggtct ctgacttcag aaaggacttc caattctaca aggtcagaga aatcaacaac     2940

taccaccacg cccacgacgc ctacttgaac gccgtcgtcg gtaccgcctt gatcaagaag     3000

tacccaaagt tggaatctga attcgtctac ggtgactaca aggtctacga cgtcagaaag     3060

atgatcgcca agtctgaaca agaaatcggt aaggccaccg ccaagtactt cttctactct     3120

aacatcatga acttcttcaa gaccgaaatc accttggcca acggtgaaat cagaaagaga     3180

ccattgatcg aaaccaacgg tgaaaccggt gaaatcgtct gggacaaggg tagagacttc     3240

gccaccgtca gaaaggtctt gtctatgcca caagtcaaca tcgtcaagaa gaccgaagtc     3300

caaaccggtg gtttctctaa ggaatctatc ttgccaaaga gaaactctga caagttgatc     3360

gccagaaaga aggactggga cccaaagaag tacggtggtt tcgactctcc aaccgtcgcc     3420

tactctgtct tggtcgtcgc caaggtcgaa aagggtaagt ctaagaagtt gaagtctgtc     3480

aaggaattgt tgggtatcac catcatggaa agatcttctt tcgaaaagaa cccaatcgac     3540

ttcttggaag ccaagggtta caaggaagtc aagaaggact tgatcatcaa gttgccaaag     3600

tactctttgt tcgaattgga aaacggtaga aagagaatgt tggcctctgc cggtgaattg     3660

caaaagggta acgaattggc cttgccatct aagtacgtca acttcttgta cttggcctct     3720

cactacgaaa agttgaaggg ttctccagaa gacaacgaac aaaagcaatt gttcgtcgaa     3780

caacacaagc actacttgga cgaaatcatc gaacaaatct ctgaattctc taagagagtc     3840

atcttggccg acgccaactt ggacaaggtc ttgtctgcct acaacaagca cagagacaag     3900

ccaatcagag aacaagccga aaacatcatc cacttgttca ccttgaccaa cttgggtgcc     3960

ccagccgcct tcaagtactt cgacaccacc atcgacagaa agagatacac ctctaccaag     4020

gaagtcttgg acgccacctt gatccaccaa tctatcaccg gtttgtacga aaccagaatc     4080

gacttgtctc aattgggtgg tgacggtggt ggttctccaa agaagaagag aaaggtctaa     4140


<210>  108
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using U rich codons of Table 4, with start 
       and stop codons

<400>  108
atggataaaa aatattctat tggtttagat attggtacta attctgttgg ttgggctgtt       60

attactgatg aatataaagt tccttctaaa aaatttaaag ttttaggtaa tactgatcgt      120

cattctatta aaaaaaattt aattggtgct ttattatttg attctggtga aactgctgaa      180

gctactcgtt taaaacgtac tgctcgtcgt cgttatactc gtcgtaaaaa tcgtatttgt      240

tatttacaag aaattttttc taatgaaatg gctaaagttg atgattcttt ttttcatcgt      300

ttagaagaat cttttttagt tgaagaagat aaaaaacatg aacgtcatcc tatttttggt      360

aatattgttg atgaagttgc ttatcatgaa aaatatccta ctatttatca tttacgtaaa      420

aaattagttg attctactga taaagctgat ttacgtttaa tttatttagc tttagctcat      480

atgattaaat ttcgtggtca ttttttaatt gaaggtgatt taaatcctga taattctgat      540

gttgataaat tatttattca attagttcaa acttataatc aattatttga agaaaatcct      600

attaatgctt ctggtgttga tgctaaagct attttatctg ctcgtttatc taaatctcgt      660

cgtttagaaa atttaattgc tcaattacct ggtgaaaaaa aaaatggttt atttggtaat      720

ttaattgctt tatctttagg tttaactcct aattttaaat ctaattttga tttagctgaa      780

gatgctaaat tacaattatc taaagatact tatgatgatg atttagataa tttattagct      840

caaattggtg atcaatatgc tgatttattt ttagctgcta aaaatttatc tgatgctatt      900

ttattatctg atattttacg tgttaatact gaaattacta aagctccttt atctgcttct      960

atgattaaac gttatgatga acatcatcaa gatttaactt tattaaaagc tttagttcgt     1020

caacaattac ctgaaaaata taaagaaatt ttttttgatc aatctaaaaa tggttatgct     1080

ggttatattg atggtggtgc ttctcaagaa gaattttata aatttattaa acctatttta     1140

gaaaaaatgg atggtactga agaattatta gttaaattaa atcgtgaaga tttattacgt     1200

aaacaacgta cttttgataa tggttctatt cctcatcaaa ttcatttagg tgaattacat     1260

gctattttac gtcgtcaaga agatttttat ccttttttaa aagataatcg tgaaaaaatt     1320

gaaaaaattt taacttttcg tattccttat tatgttggtc ctttagctcg tggtaattct     1380

cgttttgctt ggatgactcg taaatctgaa gaaactatta ctccttggaa ttttgaagaa     1440

gttgttgata aaggtgcttc tgctcaatct tttattgaac gtatgactaa ttttgataaa     1500

aatttaccta atgaaaaagt tttacctaaa cattctttat tatatgaata ttttactgtt     1560

tataatgaat taactaaagt taaatatgtt actgaaggta tgcgtaaacc tgctttttta     1620

tctggtgaac aaaaaaaagc tattgttgat ttattattta aaactaatcg taaagttact     1680

gttaaacaat taaaagaaga ttattttaaa aaaattgaat gttttgattc tgttgaaatt     1740

tctggtgttg aagatcgttt taatgcttct ttaggtactt atcatgattt attaaaaatt     1800

attaaagata aagatttttt agataatgaa gaaaatgaag atattttaga agatattgtt     1860

ttaactttaa ctttatttga agatcgtgaa atgattgaag aacgtttaaa aacttatgct     1920

catttatttg atgataaagt tatgaaacaa ttaaaacgtc gtcgttatac tggttggggt     1980

cgtttatctc gtaaattaat taatggtatt cgtgataaac aatctggtaa aactatttta     2040

gattttttaa aatctgatgg ttttgctaat cgtaatttta tgcaattaat tcatgatgat     2100

tctttaactt ttaaagaaga tattcaaaaa gctcaagttt ctggtcaagg tgattcttta     2160

catgaacata ttgctaattt agctggttct cctgctatta aaaaaggtat tttacaaact     2220

gttaaagttg ttgatgaatt agttaaagtt atgggtcgtc ataaacctga aaatattgtt     2280

attgaaatgg ctcgtgaaaa tcaaactact caaaaaggtc aaaaaaattc tcgtgaacgt     2340

atgaaacgta ttgaagaagg tattaaagaa ttaggttctc aaattttaaa agaacatcct     2400

gttgaaaata ctcaattaca aaatgaaaaa ttatatttat attatttaca aaatggtcgt     2460

gatatgtatg ttgatcaaga attagatatt aatcgtttat ctgattatga tgttgatcat     2520

attgttcctc aatctttttt aaaagatgat tctattgata ataaagtttt aactcgttct     2580

gataaaaatc gtggtaaatc tgataatgtt ccttctgaag aagttgttaa aaaaatgaaa     2640

aattattggc gtcaattatt aaatgctaaa ttaattactc aacgtaaatt tgataattta     2700

actaaagctg aacgtggtgg tttatctgaa ttagataaag ctggttttat taaacgtcaa     2760

ttagttgaaa ctcgtcaaat tactaaacat gttgctcaaa ttttagattc tcgtatgaat     2820

actaaatatg atgaaaatga taaattaatt cgtgaagtta aagttattac tttaaaatct     2880

aaattagttt ctgattttcg taaagatttt caattttata aagttcgtga aattaataat     2940

tatcatcatg ctcatgatgc ttatttaaat gctgttgttg gtactgcttt aattaaaaaa     3000

tatcctaaat tagaatctga atttgtttat ggtgattata aagtttatga tgttcgtaaa     3060

atgattgcta aatctgaaca agaaattggt aaagctactg ctaaatattt tttttattct     3120

aatattatga atttttttaa aactgaaatt actttagcta atggtgaaat tcgtaaacgt     3180

cctttaattg aaactaatgg tgaaactggt gaaattgttt gggataaagg tcgtgatttt     3240

gctactgttc gtaaagtttt atctatgcct caagttaata ttgttaaaaa aactgaagtt     3300

caaactggtg gtttttctaa agaatctatt ttacctaaac gtaattctga taaattaatt     3360

gctcgtaaaa aagattggga tcctaaaaaa tatggtggtt ttgattctcc tactgttgct     3420

tattctgttt tagttgttgc taaagttgaa aaaggtaaat ctaaaaaatt aaaatctgtt     3480

aaagaattat taggtattac tattatggaa cgttcttctt ttgaaaaaaa tcctattgat     3540

tttttagaag ctaaaggtta taaagaagtt aaaaaagatt taattattaa attacctaaa     3600

tattctttat ttgaattaga aaatggtcgt aaacgtatgt tagcttctgc tggtgaatta     3660

caaaaaggta atgaattagc tttaccttct aaatatgtta attttttata tttagcttct     3720

cattatgaaa aattaaaagg ttctcctgaa gataatgaac aaaaacaatt atttgttgaa     3780

caacataaac attatttaga tgaaattatt gaacaaattt ctgaattttc taaacgtgtt     3840

attttagctg atgctaattt agataaagtt ttatctgctt ataataaaca tcgtgataaa     3900

cctattcgtg aacaagctga aaatattatt catttattta ctttaactaa tttaggtgct     3960

cctgctgctt ttaaatattt tgatactact attgatcgta aacgttatac ttctactaaa     4020

gaagttttag atgctacttt aattcatcaa tctattactg gtttatatga aactcgtatt     4080

gatttatctc aattaggtgg tgatggtggt ggttctccta aaaaaaaacg taaagtttga     4140


<210>  109
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low G codons of Table 4, with start and
       stop codons

<400>  109
atggacaaaa aatactccat cggcctcgac atcggcacca actccgtcgg ctgggccgtc       60

atcaccgacg aatacaaagt cccctccaaa aaattcaaag tcctcggcaa caccgacaga      120

cactccatca aaaaaaacct catcggcgcc ctcctcttcg actccggcga aaccgccgaa      180

gccaccagac tcaaaagaac cgccagaaga agatacacca gaagaaaaaa cagaatctgc      240

tacctccaag aaatcttctc caacgaaatg gccaaagtcg acgactcctt cttccacaga      300

ctcgaagaat ccttcctcgt cgaagaagac aaaaaacacg aaagacaccc catcttcggc      360

aacatcgtcg acgaagtcgc ctaccacgaa aaatacccca ccatctacca cctcagaaaa      420

aaactcgtcg actccaccga caaagccgac ctcagactca tctacctcgc cctcgcccac      480

atgatcaaat tcagaggcca cttcctcatc gaaggcgacc tcaaccccga caactccgac      540

gtcgacaaac tcttcatcca actcgtccaa acctacaacc aactcttcga agaaaacccc      600

atcaacgcct ccggcgtcga cgccaaagcc atcctctccg ccagactctc caaatccaga      660

agactcgaaa acctcatcgc ccaactcccc ggcgaaaaaa aaaacggcct cttcggcaac      720

ctcatcgccc tctccctcgg cctcaccccc aacttcaaat ccaacttcga cctcgccgaa      780

gacgccaaac tccaactctc caaagacacc tacgacgacg acctcgacaa cctcctcgcc      840

caaatcggcg accaatacgc cgacctcttc ctcgccgcca aaaacctctc cgacgccatc      900

ctcctctccg acatcctcag agtcaacacc gaaatcacca aagcccccct ctccgcctcc      960

atgatcaaaa gatacgacga acaccaccaa gacctcaccc tcctcaaagc cctcgtcaga     1020

caacaactcc ccgaaaaata caaagaaatc ttcttcgacc aatccaaaaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaagaa gaattctaca aattcatcaa acccatcctc     1140

gaaaaaatgg acggcaccga agaactcctc gtcaaactca acagagaaga cctcctcaga     1200

aaacaaagaa ccttcgacaa cggctccatc ccccaccaaa tccacctcgg cgaactccac     1260

gccatcctca gaagacaaga agacttctac cccttcctca aagacaacag agaaaaaatc     1320

gaaaaaatcc tcaccttcag aatcccctac tacgtcggcc ccctcgccag aggcaactcc     1380

agattcgcct ggatgaccag aaaatccgaa gaaaccatca ccccctggaa cttcgaagaa     1440

gtcgtcgaca aaggcgcctc cgcccaatcc ttcatcgaaa gaatgaccaa cttcgacaaa     1500

aacctcccca acgaaaaagt cctccccaaa cactccctcc tctacgaata cttcaccgtc     1560

tacaacgaac tcaccaaagt caaatacgtc accgaaggca tgagaaaacc cgccttcctc     1620

tccggcgaac aaaaaaaagc catcgtcgac ctcctcttca aaaccaacag aaaagtcacc     1680

gtcaaacaac tcaaagaaga ctacttcaaa aaaatcgaat gcttcgactc cgtcgaaatc     1740

tccggcgtcg aagacagatt caacgcctcc ctcggcacct accacgacct cctcaaaatc     1800

atcaaagaca aagacttcct cgacaacgaa gaaaacgaag acatcctcga agacatcgtc     1860

ctcaccctca ccctcttcga agacagagaa atgatcgaag aaagactcaa aacctacgcc     1920

cacctcttcg acgacaaagt catgaaacaa ctcaaaagaa gaagatacac cggctggggc     1980

agactctcca gaaaactcat caacggcatc agagacaaac aatccggcaa aaccatcctc     2040

gacttcctca aatccgacgg cttcgccaac agaaacttca tgcaactcat ccacgacgac     2100

tccctcacct tcaaagaaga catccaaaaa gcccaagtct ccggccaagg cgactccctc     2160

cacgaacaca tcgccaacct cgccggctcc cccgccatca aaaaaggcat cctccaaacc     2220

gtcaaagtcg tcgacgaact cgtcaaagtc atgggcagac acaaacccga aaacatcgtc     2280

atcgaaatgg ccagagaaaa ccaaaccacc caaaaaggcc aaaaaaactc cagagaaaga     2340

atgaaaagaa tcgaagaagg catcaaagaa ctcggctccc aaatcctcaa agaacacccc     2400

gtcgaaaaca cccaactcca aaacgaaaaa ctctacctct actacctcca aaacggcaga     2460

gacatgtacg tcgaccaaga actcgacatc aacagactct ccgactacga cgtcgaccac     2520

atcgtccccc aatccttcct caaagacgac tccatcgaca acaaagtcct caccagatcc     2580

gacaaaaaca gaggcaaatc cgacaacgtc ccctccgaag aagtcgtcaa aaaaatgaaa     2640

aactactgga gacaactcct caacgccaaa ctcatcaccc aaagaaaatt cgacaacctc     2700

accaaagccg aaagaggcgg cctctccgaa ctcgacaaag ccggcttcat caaaagacaa     2760

ctcgtcgaaa ccagacaaat caccaaacac gtcgcccaaa tcctcgactc cagaatgaac     2820

accaaatacg acgaaaacga caaactcatc agagaagtca aagtcatcac cctcaaatcc     2880

aaactcgtct ccgacttcag aaaagacttc caattctaca aagtcagaga aatcaacaac     2940

taccaccacg cccacgacgc ctacctcaac gccgtcgtcg gcaccgccct catcaaaaaa     3000

taccccaaac tcgaatccga attcgtctac ggcgactaca aagtctacga cgtcagaaaa     3060

atgatcgcca aatccgaaca agaaatcggc aaagccaccg ccaaatactt cttctactcc     3120

aacatcatga acttcttcaa aaccgaaatc accctcgcca acggcgaaat cagaaaaaga     3180

cccctcatcg aaaccaacgg cgaaaccggc gaaatcgtct gggacaaagg cagagacttc     3240

gccaccgtca gaaaagtcct ctccatgccc caagtcaaca tcgtcaaaaa aaccgaagtc     3300

caaaccggcg gcttctccaa agaatccatc ctccccaaaa gaaactccga caaactcatc     3360

gccagaaaaa aagactggga ccccaaaaaa tacggcggct tcgactcccc caccgtcgcc     3420

tactccgtcc tcgtcgtcgc caaagtcgaa aaaggcaaat ccaaaaaact caaatccgtc     3480

aaagaactcc tcggcatcac catcatggaa agatcctcct tcgaaaaaaa ccccatcgac     3540

ttcctcgaag ccaaaggcta caaagaagtc aaaaaagacc tcatcatcaa actccccaaa     3600

tactccctct tcgaactcga aaacggcaga aaaagaatgc tcgcctccgc cggcgaactc     3660

caaaaaggca acgaactcgc cctcccctcc aaatacgtca acttcctcta cctcgcctcc     3720

cactacgaaa aactcaaagg ctcccccgaa gacaacgaac aaaaacaact cttcgtcgaa     3780

caacacaaac actacctcga cgaaatcatc gaacaaatct ccgaattctc caaaagagtc     3840

atcctcgccg acgccaacct cgacaaagtc ctctccgcct acaacaaaca cagagacaaa     3900

cccatcagag aacaagccga aaacatcatc cacctcttca ccctcaccaa cctcggcgcc     3960

cccgccgcct tcaaatactt cgacaccacc atcgacagaa aaagatacac ctccaccaaa     4020

gaagtcctcg acgccaccct catccaccaa tccatcaccg gcctctacga aaccagaatc     4080

gacctctccc aactcggcgg cgacggcggc ggctccccca aaaaaaaaag aaaagtctga     4140


<210>  110
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low C codons of Table 4, with start and
       stop codons

<400>  110
atggataaga agtatagtat tggattggat attggaacaa atagtgtggg atgggctgtg       60

attacagatg agtataaggt gcctagtaag aagtttaagg tgttgggaaa tacagataga      120

catagtatta agaagaattt gattggagct ttgttgtttg atagtggaga gacagctgag      180

gctacaagat tgaagagaac agctagaaga agatatacaa gaagaaagaa tagaatttgt      240

tatttgcagg agatttttag taatgagatg gctaaggtgg atgatagttt ttttcataga      300

ttggaggaga gttttttggt ggaggaggat aagaagcatg agagacatcc tatttttgga      360

aatattgtgg atgaggtggc ttatcatgag aagtatccta caatttatca tttgagaaag      420

aagttggtgg atagtacaga taaggctgat ttgagattga tttatttggc tttggctcat      480

atgattaagt ttagaggaca ttttttgatt gagggagatt tgaatcctga taatagtgat      540

gtggataagt tgtttattca gttggtgcag acatataatc agttgtttga ggagaatcct      600

attaatgcta gtggagtgga tgctaaggct attttgagtg ctagattgag taagagtaga      660

agattggaga atttgattgc tcagttgcct ggagagaaga agaatggatt gtttggaaat      720

ttgattgctt tgagtttggg attgacacct aattttaaga gtaattttga tttggctgag      780

gatgctaagt tgcagttgag taaggataca tatgatgatg atttggataa tttgttggct      840

cagattggag atcagtatgc tgatttgttt ttggctgcta agaatttgag tgatgctatt      900

ttgttgagtg atattttgag agtgaataca gagattacaa aggctccttt gagtgctagt      960

atgattaaga gatatgatga gcatcatcag gatttgacat tgttgaaggc tttggtgaga     1020

cagcagttgc ctgagaagta taaggagatt ttttttgatc agagtaagaa tggatatgct     1080

ggatatattg atggaggagc tagtcaggag gagttttata agtttattaa gcctattttg     1140

gagaagatgg atggaacaga ggagttgttg gtgaagttga atagagagga tttgttgaga     1200

aagcagagaa catttgataa tggaagtatt cctcatcaga ttcatttggg agagttgcat     1260

gctattttga gaagacagga ggatttttat ccttttttga aggataatag agagaagatt     1320

gagaagattt tgacatttag aattccttat tatgtgggac ctttggctag aggaaatagt     1380

agatttgctt ggatgacaag aaagagtgag gagacaatta caccttggaa ttttgaggag     1440

gtggtggata agggagctag tgctcagagt tttattgaga gaatgacaaa ttttgataag     1500

aatttgccta atgagaaggt gttgcctaag catagtttgt tgtatgagta ttttacagtg     1560

tataatgagt tgacaaaggt gaagtatgtg acagagggaa tgagaaagcc tgcttttttg     1620

agtggagagc agaagaaggc tattgtggat ttgttgttta agacaaatag aaaggtgaca     1680

gtgaagcagt tgaaggagga ttattttaag aagattgagt gttttgatag tgtggagatt     1740

agtggagtgg aggatagatt taatgctagt ttgggaacat atcatgattt gttgaagatt     1800

attaaggata aggatttttt ggataatgag gagaatgagg atattttgga ggatattgtg     1860

ttgacattga cattgtttga ggatagagag atgattgagg agagattgaa gacatatgct     1920

catttgtttg atgataaggt gatgaagcag ttgaagagaa gaagatatac aggatgggga     1980

agattgagta gaaagttgat taatggaatt agagataagc agagtggaaa gacaattttg     2040

gattttttga agagtgatgg atttgctaat agaaatttta tgcagttgat tcatgatgat     2100

agtttgacat ttaaggagga tattcagaag gctcaggtga gtggacaggg agatagtttg     2160

catgagcata ttgctaattt ggctggaagt cctgctatta agaagggaat tttgcagaca     2220

gtgaaggtgg tggatgagtt ggtgaaggtg atgggaagac ataagcctga gaatattgtg     2280

attgagatgg ctagagagaa tcagacaaca cagaagggac agaagaatag tagagagaga     2340

atgaagagaa ttgaggaggg aattaaggag ttgggaagtc agattttgaa ggagcatcct     2400

gtggagaata cacagttgca gaatgagaag ttgtatttgt attatttgca gaatggaaga     2460

gatatgtatg tggatcagga gttggatatt aatagattga gtgattatga tgtggatcat     2520

attgtgcctc agagtttttt gaaggatgat agtattgata ataaggtgtt gacaagaagt     2580

gataagaata gaggaaagag tgataatgtg cctagtgagg aggtggtgaa gaagatgaag     2640

aattattgga gacagttgtt gaatgctaag ttgattacac agagaaagtt tgataatttg     2700

acaaaggctg agagaggagg attgagtgag ttggataagg ctggatttat taagagacag     2760

ttggtggaga caagacagat tacaaagcat gtggctcaga ttttggatag tagaatgaat     2820

acaaagtatg atgagaatga taagttgatt agagaggtga aggtgattac attgaagagt     2880

aagttggtga gtgattttag aaaggatttt cagttttata aggtgagaga gattaataat     2940

tatcatcatg ctcatgatgc ttatttgaat gctgtggtgg gaacagcttt gattaagaag     3000

tatcctaagt tggagagtga gtttgtgtat ggagattata aggtgtatga tgtgagaaag     3060

atgattgcta agagtgagca ggagattgga aaggctacag ctaagtattt tttttatagt     3120

aatattatga atttttttaa gacagagatt acattggcta atggagagat tagaaagaga     3180

cctttgattg agacaaatgg agagacagga gagattgtgt gggataaggg aagagatttt     3240

gctacagtga gaaaggtgtt gagtatgcct caggtgaata ttgtgaagaa gacagaggtg     3300

cagacaggag gatttagtaa ggagagtatt ttgcctaaga gaaatagtga taagttgatt     3360

gctagaaaga aggattggga tcctaagaag tatggaggat ttgatagtcc tacagtggct     3420

tatagtgtgt tggtggtggc taaggtggag aagggaaaga gtaagaagtt gaagagtgtg     3480

aaggagttgt tgggaattac aattatggag agaagtagtt ttgagaagaa tcctattgat     3540

tttttggagg ctaagggata taaggaggtg aagaaggatt tgattattaa gttgcctaag     3600

tatagtttgt ttgagttgga gaatggaaga aagagaatgt tggctagtgc tggagagttg     3660

cagaagggaa atgagttggc tttgcctagt aagtatgtga attttttgta tttggctagt     3720

cattatgaga agttgaaggg aagtcctgag gataatgagc agaagcagtt gtttgtggag     3780

cagcataagc attatttgga tgagattatt gagcagatta gtgagtttag taagagagtg     3840

attttggctg atgctaattt ggataaggtg ttgagtgctt ataataagca tagagataag     3900

cctattagag agcaggctga gaatattatt catttgttta cattgacaaa tttgggagct     3960

cctgctgctt ttaagtattt tgatacaaca attgatagaa agagatatac aagtacaaag     4020

gaggtgttgg atgctacatt gattcatcag agtattacag gattgtatga gacaagaatt     4080

gatttgagtc agttgggagg agatggagga ggaagtccta agaagaagag aaaggtgtga     4140


<210>  111
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A codons of Table 4, with start and
       stop codons

<400>  111
atggacaaga agtactccat cggcctggac atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggaccac     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgacggcggc ggctccccca agaagaagcg gaaggtgtga     4140


<210>  112
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A/U codons of Table 4, with start 
       and stop codons

<400>  112
atggacaaga agtacagcat cggcctggac atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgacggcggc ggcagcccca agaagaagcg gaaggtgtga     4140


<210>  113
<211>  4179
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A codons of Table 4, with two 
       C-terminal NLS sequences and start and stop codons

<400>  113
atggacaaga agtactccat cggcctggac atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggaccac     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgacggctcc ggctccccca agaagaagcg gaaggtggac     4140

ggctccccca agaagaagcg gaaggtggac tccggctga                            4179


<210>  114
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A codons of Table 4, with 
       start and stop codons

<400>  114
atggacaaga agtactccat cggcctggcc atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggaccac     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgacggcggc ggctccccca agaagaagcg gaaggtgtga     4140


<210>  115
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A codons of Table 4, with 
       start and stop codons and no NLS

<400>  115
atggacaaga agtactccat cggcctggcc atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggaccac     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgactga                                         4107


<210>  116
<211>  4179
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A codons of Table 4, with 
       two C-terminal NLS sequences and start and stop codons

<400>  116
atggacaaga agtactccat cggcctggcc atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggaccac     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgacggctcc ggctccccca agaagaagcg gaaggtggac     4140

ggctccccca agaagaagcg gaaggtggac tccggctga                            4179


<210>  117
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A codons of Table 4, with start 
       and stop codons

<400>  117
atggacaaga agtactccat cggcctggcc atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggacgcc     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgacggcggc ggctccccca agaagaagcg gaaggtgtga     4140


<210>  118
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A codons of Table 4, with start 
       and stop codons and no NLS

<400>  118
atggacaaga agtactccat cggcctggcc atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggacgcc     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgactga                                         4107


<210>  119
<211>  4179
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A codons of Table 4, with two 
       C-terminal NLS sequences and start and stop codons

<400>  119
atggacaaga agtactccat cggcctggcc atcggcacca actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccctccaag aagttcaagg tgctgggcaa caccgaccgg      120

cactccatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttctc caacgagatg gccaaggtgg acgactcctt cttccaccgg      300

ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg actccaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caactccgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcct ccggcgtgga cgccaaggcc atcctgtccg cccggctgtc caagtcccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgtccctggg cctgaccccc aacttcaagt ccaacttcga cctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgtc cgacgccatc      900

ctgctgtccg acatcctgcg ggtgaacacc gagatcacca aggcccccct gtccgcctcc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agtccaagaa cggctacgcc     1080

ggctacatcg acggcggcgc ctcccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggctccatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaactcc     1380

cggttcgcct ggatgacccg gaagtccgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgcctc cgcccagtcc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cactccctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

tccggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgactc cgtggagatc     1740

tccggcgtgg aggaccggtt caacgcctcc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgtccc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg     2040

gacttcctga agtccgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

tccctgacct tcaaggagga catccagaag gcccaggtgt ccggccaggg cgactccctg     2160

cacgagcaca tcgccaacct ggccggctcc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaactc ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggctccc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctgt ccgactacga cgtggacgcc     2520

atcgtgcccc agtccttcct gaaggacgac tccatcgaca acaaggtgct gacccggtcc     2580

gacaagaacc ggggcaagtc cgacaacgtg ccctccgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggactc ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagtcc     2880

aagctggtgt ccgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagtccga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agtccgagca ggagatcggc aaggccaccg ccaagtactt cttctactcc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gtccatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttctccaa ggagtccatc ctgcccaagc ggaactccga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgactcccc caccgtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaagt ccaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cggtcctcct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tactccctgt tcgagctgga gaacggccgg aagcggatgc tggcctccgc cggcgagctg     3660

cagaagggca acgagctggc cctgccctcc aagtacgtga acttcctgta cctggcctcc     3720

cactacgaga agctgaaggg ctcccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatct ccgagttctc caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgtccgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac ctccaccaag     4020

gaggtgctgg acgccaccct gatccaccag tccatcaccg gcctgtacga gacccggatc     4080

gacctgtccc agctgggcgg cgacggctcc ggctccccca agaagaagcg gaaggtggac     4140

ggctccccca agaagaagcg gaaggtggac tccggctga                            4179


<210>  120
<211>  4179
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A/U codons of Table 4, with two 
       C-terminal NLS sequences and start and stop codons

<400>  120
atggacaaga agtacagcat cggcctggac atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgacggcagc ggcagcccca agaagaagcg gaaggtggac     4140

ggcagcccca agaagaagcg gaaggtggac agcggctga                            4179


<210>  121
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A/U codons of Table 4, with start 
       and stop codons and no NLS

<400>  121
atggacaaga agtacagcat cggcctggac atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgactga                                         4107


<210>  122
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A/U codons of Table 4, with
       start and stop codons

<400>  122
atggacaaga agtacagcat cggcctggcc atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgacggcggc ggcagcccca agaagaagcg gaaggtgtga     4140


<210>  123
<211>  4179
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A/U codons of Table 4, with
       two C-terminal NLS sequences and start and stop codons

<400>  123
atggacaaga agtacagcat cggcctggcc atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgacggcagc ggcagcccca agaagaagcg gaaggtggac     4140

ggcagcccca agaagaagcg gaaggtggac agcggctga                            4179


<210>  124
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A/U codons of Table 4, with
       start and stop codons and no NLS

<400>  124
atggacaaga agtacagcat cggcctggcc atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggaccac     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgactga                                         4107


<210>  125
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A/U codons of Table 4, with start 
       and stop codons

<400>  125
atggacaaga agtacagcat cggcctggcc atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggacgcc     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgacggcggc ggcagcccca agaagaagcg gaaggtgtga     4140


<210>  126
<211>  4179
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A/U codons of Table 4, with two 
       C-terminal NLS sequences and start and stop codons

<400>  126
atggacaaga agtacagcat cggcctggcc atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggacgcc     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgacggcagc ggcagcccca agaagaagcg gaaggtggac     4140

ggcagcccca agaagaagcg gaaggtggac agcggctga                            4179


<210>  127
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A/U codons of Table 4, with start 
       and stop codons and no NLS

<400>  127
atggacaaga agtacagcat cggcctggcc atcggcacca acagcgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aagttcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggcga gaccgccgag      180

gccacccggc tgaagcggac cgcccggcgg cggtacaccc ggcggaagaa ccggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgacagctt cttccaccgg      300

ctggaggaga gcttcctggt ggaggaggac aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgcggaag      420

aagctggtgg acagcaccga caaggccgac ctgcggctga tctacctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgagcg cccggctgag caagagccgg      660

cggctggaga acctgatcgc ccagctgccc ggcgagaaga agaacggcct gttcggcaac      720

ctgatcgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gacgccaagc tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttc ctggccgcca agaacctgag cgacgccatc      900

ctgctgagcg acatcctgcg ggtgaacacc gagatcacca aggcccccct gagcgccagc      960

atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc cctggtgcgg     1020

cagcagctgc ccgagaagta caaggagatc ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacatcg acggcggcgc cagccaggag gagttctaca agttcatcaa gcccatcctg     1140

gagaagatgg acggcaccga ggagctgctg gtgaagctga accgggagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg cgagctgcac     1260

gccatcctgc ggcggcagga ggacttctac cccttcctga aggacaaccg ggagaagatc     1320

gagaagatcc tgaccttccg gatcccctac tacgtgggcc ccctggcccg gggcaacagc     1380

cggttcgcct ggatgacccg gaagagcgag gagaccatca ccccctggaa cttcgaggag     1440

gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgacaag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tacaacgagc tgaccaaggt gaagtacgtg accgagggca tgcggaagcc cgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttca agaccaaccg gaaggtgacc     1680

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatc     1740

agcggcgtgg aggaccggtt caacgccagc ctgggcacct accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtg     1860

ctgaccctga ccctgttcga ggaccgggag atgatcgagg agcggctgaa gacctacgcc     1920

cacctgttcg acgacaaggt gatgaagcag ctgaagcggc ggcggtacac cggctggggc     1980

cggctgagcc ggaagctgat caacggcatc cgggacaagc agagcggcaa gaccatcctg     2040

gacttcctga agagcgacgg cttcgccaac cggaacttca tgcagctgat ccacgacgac     2100

agcctgacct tcaaggagga catccagaag gcccaggtga gcggccaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggcagc cccgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaggtg atgggccggc acaagcccga gaacatcgtg     2280

atcgagatgg cccgggagaa ccagaccacc cagaagggcc agaagaacag ccgggagcgg     2340

atgaagcgga tcgaggaggg catcaaggag ctgggcagcc agatcctgaa ggagcacccc     2400

gtggagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgg     2460

gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggacgcc     2520

atcgtgcccc agagcttcct gaaggacgac agcatcgaca acaaggtgct gacccggagc     2580

gacaagaacc ggggcaagag cgacaacgtg cccagcgagg aggtggtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgatcaccc agcggaagtt cgacaacctg     2700

accaaggccg agcggggcgg cctgagcgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga cccggcagat caccaagcac gtggcccaga tcctggacag ccggatgaac     2820

accaagtacg acgagaacga caagctgatc cgggaggtga aggtgatcac cctgaagagc     2880

aagctggtga gcgacttccg gaaggacttc cagttctaca aggtgcggga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtggtgg gcaccgccct gatcaagaag     3000

taccccaagc tggagagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctacagc     3120

aacatcatga acttcttcaa gaccgagatc accctggcca acggcgagat ccggaagcgg     3180

cccctgatcg agaccaacgg cgagaccggc gagatcgtgt gggacaaggg ccgggacttc     3240

gccaccgtgc ggaaggtgct gagcatgccc caggtgaaca tcgtgaagaa gaccgaggtg     3300

cagaccggcg gcttcagcaa ggagagcatc ctgcccaagc ggaacagcga caagctgatc     3360

gcccggaaga aggactggga ccccaagaag tacggcggct tcgacagccc caccgtggcc     3420

tacagcgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagagcgtg     3480

aaggagctgc tgggcatcac catcatggag cggagcagct tcgagaagaa ccccatcgac     3540

ttcctggagg ccaagggcta caaggaggtg aagaaggacc tgatcatcaa gctgcccaag     3600

tacagcctgt tcgagctgga gaacggccgg aagcggatgc tggccagcgc cggcgagctg     3660

cagaagggca acgagctggc cctgcccagc aagtacgtga acttcctgta cctggccagc     3720

cactacgaga agctgaaggg cagccccgag gacaacgagc agaagcagct gttcgtggag     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgggtg     3840

atcctggccg acgccaacct ggacaaggtg ctgagcgcct acaacaagca ccgggacaag     3900

cccatccggg agcaggccga gaacatcatc cacctgttca ccctgaccaa cctgggcgcc     3960

cccgccgcct tcaagtactt cgacaccacc atcgaccgga agcggtacac cagcaccaag     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacccggatc     4080

gacctgagcc agctgggcgg cgactga                                         4107


<210>  128
<211>  3312
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Nme Cas9 ORF using low A codons of Table 4, with start
       and stop codons

<400>  128
atggccgcct tcaagcccaa ctccatcaac tacatcctgg gcctggacat cggcatcgcc       60

tccgtgggct gggccatggt ggagatcgac gaggaggaga accccatccg gctgatcgac      120

ctgggcgtgc gggtgttcga gcgggccgag gtgcccaaga ccggcgactc cctggccatg      180

gcccggcggc tggcccggtc cgtgcggcgg ctgacccggc ggcgggccca ccggctgctg      240

cggacccggc ggctgctgaa gcgggagggc gtgctgcagg ccgccaactt cgacgagaac      300

ggcctgatca agtccctgcc caacaccccc tggcagctgc gggccgccgc cctggaccgg      360

aagctgaccc ccctggagtg gtccgccgtg ctgctgcacc tgatcaagca ccggggctac      420

ctgtcccagc ggaagaacga gggcgagacc gccgacaagg agctgggcgc cctgctgaag      480

ggcgtggccg gcaacgccca cgccctgcag accggcgact tccggacccc cgccgagctg      540

gccctgaaca agttcgagaa ggagtccggc cacatccgga accagcggtc cgactactcc      600

cacaccttct cccggaagga cctgcaggcc gagctgatcc tgctgttcga gaagcagaag      660

gagttcggca acccccacgt gtccggcggc ctgaaggagg gcatcgagac cctgctgatg      720

acccagcggc ccgccctgtc cggcgacgcc gtgcagaaga tgctgggcca ctgcaccttc      780

gagcccgccg agcccaaggc cgccaagaac acctacaccg ccgagcggtt catctggctg      840

accaagctga acaacctgcg gatcctggag cagggctccg agcggcccct gaccgacacc      900

gagcgggcca ccctgatgga cgagccctac cggaagtcca agctgaccta cgcccaggcc      960

cggaagctgc tgggcctgga ggacaccgcc ttcttcaagg gcctgcggta cggcaaggac     1020

aacgccgagg cctccaccct gatggagatg aaggcctacc acgccatctc ccgggccctg     1080

gagaaggagg gcctgaagga caagaagtcc cccctgaacc tgtcccccga gctgcaggac     1140

gagatcggca ccgccttctc cctgttcaag accgacgagg acatcaccgg ccggctgaag     1200

gaccggatcc agcccgagat cctggaggcc ctgctgaagc acatctcctt cgacaagttc     1260

gtgcagatct ccctgaaggc cctgcggcgg atcgtgcccc tgatggagca gggcaagcgg     1320

tacgacgagg cctgcgccga gatctacggc gaccactacg gcaagaagaa caccgaggag     1380

aagatctacc tgccccccat ccccgccgac gagatccgga accccgtggt gctgcgggcc     1440

ctgtcccagg cccggaaggt gatcaacggc gtggtgcggc ggtacggctc ccccgcccgg     1500

atccacatcg agaccgcccg ggaggtgggc aagtccttca aggaccggaa ggagatcgag     1560

aagcggcagg aggagaaccg gaaggaccgg gagaaggccg ccgccaagtt ccgggagtac     1620

ttccccaact tcgtgggcga gcccaagtcc aaggacatcc tgaagctgcg gctgtacgag     1680

cagcagcacg gcaagtgcct gtactccggc aaggagatca acctgggccg gctgaacgag     1740

aagggctacg tggagatcga ccacgccctg cccttctccc ggacctggga cgactccttc     1800

aacaacaagg tgctggtgct gggctccgag aaccagaaca agggcaacca gaccccctac     1860

gagtacttca acggcaagga caactcccgg gagtggcagg agttcaaggc ccgggtggag     1920

acctcccggt tcccccggtc caagaagcag cggatcctgc tgcagaagtt cgacgaggac     1980

ggcttcaagg agcggaacct gaacgacacc cggtacgtga accggttcct gtgccagttc     2040

gtggccgacc ggatgcggct gaccggcaag ggcaagaagc gggtgttcgc ctccaacggc     2100

cagatcacca acctgctgcg gggcttctgg ggcctgcgga aggtgcgggc cgagaacgac     2160

cggcaccacg ccctggacgc cgtggtggtg gcctgctcca ccgtggccat gcagcagaag     2220

atcacccggt tcgtgcggta caaggagatg aacgccttcg acggcaagac catcgacaag     2280

gagaccggcg aggtgctgca ccagaagacc cacttccccc agccctggga gttcttcgcc     2340

caggaggtga tgatccgggt gttcggcaag cccgacggca agcccgagtt cgaggaggcc     2400

gacaccctgg agaagctgcg gaccctgctg gccgagaagc tgtcctcccg gcccgaggcc     2460

gtgcacgagt acgtgacccc cctgttcgtg tcccgggccc ccaaccggaa gatgtccggc     2520

cagggccaca tggagaccgt gaagtccgcc aagcggctgg acgagggcgt gtccgtgctg     2580

cgggtgcccc tgacccagct gaagctgaag gacctggaga agatggtgaa ccgggagcgg     2640

gagcccaagc tgtacgaggc cctgaaggcc cggctggagg cccacaagga cgaccccgcc     2700

aaggccttcg ccgagccctt ctacaagtac gacaaggccg gcaaccggac ccagcaggtg     2760

aaggccgtgc gggtggagca ggtgcagaag accggcgtgt gggtgcggaa ccacaacggc     2820

atcgccgaca acgccaccat ggtgcgggtg gacgtgttcg agaagggcga caagtactac     2880

ctggtgccca tctactcctg gcaggtggcc aagggcatcc tgcccgaccg ggccgtggtg     2940

cagggcaagg acgaggagga ctggcagctg atcgacgact ccttcaactt caagttctcc     3000

ctgcacccca acgacctggt ggaggtgatc accaagaagg cccggatgtt cggctacttc     3060

gcctcctgcc accggggcac cggcaacatc aacatccgga tccacgacct ggaccacaag     3120

atcggcaaga acggcatcct ggagggcatc ggcgtgaaga ccgccctgtc cttccagaag     3180

taccagatcg acgagctggg caaggagatc cggccctgcc ggctgaagaa gcggcccccc     3240

gtgcggtccg gcaagcggac cgccgacggc tccgagttcg agtcccccaa gaagaagcgg     3300

aaggtggagt ga                                                         3312


<210>  129
<211>  3312
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Nme Cas9 ORF using low A/U codons of Table 4, with 
       start and stop codons

<400>  129
atggccgcct tcaagcccaa cagcatcaac tacatcctgg gcctggacat cggcatcgcc       60

agcgtgggct gggccatggt ggagatcgac gaggaggaga accccatccg gctgatcgac      120

ctgggcgtgc gggtgttcga gcgggccgag gtgcccaaga ccggcgacag cctggccatg      180

gcccggcggc tggcccggag cgtgcggcgg ctgacccggc ggcgggccca ccggctgctg      240

cggacccggc ggctgctgaa gcgggagggc gtgctgcagg ccgccaactt cgacgagaac      300

ggcctgatca agagcctgcc caacaccccc tggcagctgc gggccgccgc cctggaccgg      360

aagctgaccc ccctggagtg gagcgccgtg ctgctgcacc tgatcaagca ccggggctac      420

ctgagccagc ggaagaacga gggcgagacc gccgacaagg agctgggcgc cctgctgaag      480

ggcgtggccg gcaacgccca cgccctgcag accggcgact tccggacccc cgccgagctg      540

gccctgaaca agttcgagaa ggagagcggc cacatccgga accagcggag cgactacagc      600

cacaccttca gccggaagga cctgcaggcc gagctgatcc tgctgttcga gaagcagaag      660

gagttcggca acccccacgt gagcggcggc ctgaaggagg gcatcgagac cctgctgatg      720

acccagcggc ccgccctgag cggcgacgcc gtgcagaaga tgctgggcca ctgcaccttc      780

gagcccgccg agcccaaggc cgccaagaac acctacaccg ccgagcggtt catctggctg      840

accaagctga acaacctgcg gatcctggag cagggcagcg agcggcccct gaccgacacc      900

gagcgggcca ccctgatgga cgagccctac cggaagagca agctgaccta cgcccaggcc      960

cggaagctgc tgggcctgga ggacaccgcc ttcttcaagg gcctgcggta cggcaaggac     1020

aacgccgagg ccagcaccct gatggagatg aaggcctacc acgccatcag ccgggccctg     1080

gagaaggagg gcctgaagga caagaagagc cccctgaacc tgagccccga gctgcaggac     1140

gagatcggca ccgccttcag cctgttcaag accgacgagg acatcaccgg ccggctgaag     1200

gaccggatcc agcccgagat cctggaggcc ctgctgaagc acatcagctt cgacaagttc     1260

gtgcagatca gcctgaaggc cctgcggcgg atcgtgcccc tgatggagca gggcaagcgg     1320

tacgacgagg cctgcgccga gatctacggc gaccactacg gcaagaagaa caccgaggag     1380

aagatctacc tgccccccat ccccgccgac gagatccgga accccgtggt gctgcgggcc     1440

ctgagccagg cccggaaggt gatcaacggc gtggtgcggc ggtacggcag ccccgcccgg     1500

atccacatcg agaccgcccg ggaggtgggc aagagcttca aggaccggaa ggagatcgag     1560

aagcggcagg aggagaaccg gaaggaccgg gagaaggccg ccgccaagtt ccgggagtac     1620

ttccccaact tcgtgggcga gcccaagagc aaggacatcc tgaagctgcg gctgtacgag     1680

cagcagcacg gcaagtgcct gtacagcggc aaggagatca acctgggccg gctgaacgag     1740

aagggctacg tggagatcga ccacgccctg cccttcagcc ggacctggga cgacagcttc     1800

aacaacaagg tgctggtgct gggcagcgag aaccagaaca agggcaacca gaccccctac     1860

gagtacttca acggcaagga caacagccgg gagtggcagg agttcaaggc ccgggtggag     1920

accagccggt tcccccggag caagaagcag cggatcctgc tgcagaagtt cgacgaggac     1980

ggcttcaagg agcggaacct gaacgacacc cggtacgtga accggttcct gtgccagttc     2040

gtggccgacc ggatgcggct gaccggcaag ggcaagaagc gggtgttcgc cagcaacggc     2100

cagatcacca acctgctgcg gggcttctgg ggcctgcgga aggtgcgggc cgagaacgac     2160

cggcaccacg ccctggacgc cgtggtggtg gcctgcagca ccgtggccat gcagcagaag     2220

atcacccggt tcgtgcggta caaggagatg aacgccttcg acggcaagac catcgacaag     2280

gagaccggcg aggtgctgca ccagaagacc cacttccccc agccctggga gttcttcgcc     2340

caggaggtga tgatccgggt gttcggcaag cccgacggca agcccgagtt cgaggaggcc     2400

gacaccctgg agaagctgcg gaccctgctg gccgagaagc tgagcagccg gcccgaggcc     2460

gtgcacgagt acgtgacccc cctgttcgtg agccgggccc ccaaccggaa gatgagcggc     2520

cagggccaca tggagaccgt gaagagcgcc aagcggctgg acgagggcgt gagcgtgctg     2580

cgggtgcccc tgacccagct gaagctgaag gacctggaga agatggtgaa ccgggagcgg     2640

gagcccaagc tgtacgaggc cctgaaggcc cggctggagg cccacaagga cgaccccgcc     2700

aaggccttcg ccgagccctt ctacaagtac gacaaggccg gcaaccggac ccagcaggtg     2760

aaggccgtgc gggtggagca ggtgcagaag accggcgtgt gggtgcggaa ccacaacggc     2820

atcgccgaca acgccaccat ggtgcgggtg gacgtgttcg agaagggcga caagtactac     2880

ctggtgccca tctacagctg gcaggtggcc aagggcatcc tgcccgaccg ggccgtggtg     2940

cagggcaagg acgaggagga ctggcagctg atcgacgaca gcttcaactt caagttcagc     3000

ctgcacccca acgacctggt ggaggtgatc accaagaagg cccggatgtt cggctacttc     3060

gccagctgcc accggggcac cggcaacatc aacatccgga tccacgacct ggaccacaag     3120

atcggcaaga acggcatcct ggagggcatc ggcgtgaaga ccgccctgag cttccagaag     3180

taccagatcg acgagctggg caaggagatc cggccctgcc ggctgaagaa gcggcccccc     3240

gtgcggagcg gcaagcggac cgccgacggc agcgagttcg agagccccaa gaagaagcgg     3300

aaggtggagt ga                                                         3312


<210>  130
<211>  4146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS1, with start and 
       stop codons

<400>  130
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcctgg cagcaaagag aagcagaaca     4140

acatag                                                                4146


<210>  131
<211>  4146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS2, with start and 
       stop codons

<400>  131
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagccagg cagcaaagag aagcagaaca     4140

acatag                                                                4146


<210>  132
<211>  4149
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS3, with start and 
       stop codons

<400>  132
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcccgg caccggcaaa gagagaaaga     4140

acaacatag                                                             4149


<210>  133
<211>  4146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS4, with start and 
       stop codons

<400>  133
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagccagg cagcaaagag accgagaaca     4140

acatag                                                                4146


<210>  134
<211>  4146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS5, with start and 
       stop codons

<400>  134
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcagag cagcaaagag accgagaaca     4140

acatag                                                                4146


<210>  135
<211>  4152
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS6, with start and 
       stop codons

<400>  135
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcgcag cagcaaagag aagctggagc     4140

atggcagcat ag                                                         4152


<210>  136
<211>  4152
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS7, with start and 
       stop codons

<400>  136
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcgcag cagcaaagag agtctggagc     4140

atggcattct ag                                                         4152


<210>  137
<211>  4152
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS8, with start and 
       stop codons

<400>  137
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcgcag cagcaaagag aagctggagc     4140

atggcattct ag                                                         4152


<210>  138
<211>  4149
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS9, with start and 
       stop codons

<400>  138
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcgcag cagcaaagag aaagtacttc     4140

gcagcatag                                                             4149


<210>  139
<211>  4149
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS10, with start and
       stop codons

<400>  139
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcagag cagcaaagag aaaggcattc     4140

gcagcatag                                                             4149


<210>  140
<211>  4149
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS11, with start and
       stop codons

<400>  140
atggacaaga agtacagcat cggactggac atcggaacaa acagcgtcgg atgggcagtc       60

atcacagacg aatacaaggt cccgagcaag aagttcaagg tcctgggaaa cacagacaga      120

cacagcatca agaagaacct gatcggagca ctgctgttcg acagcggaga aacagcagaa      180

gcaacaagac tgaagagaac agcaagaaga agatacacaa gaagaaagaa cagaatctgc      240

tacctgcagg aaatcttcag caacgaaatg gcaaaggtcg acgacagctt cttccacaga      300

ctggaagaaa gcttcctggt cgaagaagac aagaagcacg aaagacaccc gatcttcgga      360

aacatcgtcg acgaagtcgc ataccacgaa aagtacccga caatctacca cctgagaaag      420

aagctggtcg acagcacaga caaggcagac ctgagactga tctacctggc actggcacac      480

atgatcaagt tcagaggaca cttcctgatc gaaggagacc tgaacccgga caacagcgac      540

gtcgacaagc tgttcatcca gctggtccag acatacaacc agctgttcga agaaaacccg      600

atcaacgcaa gcggagtcga cgcaaaggca atcctgagcg caagactgag caagagcaga      660

agactggaaa acctgatcgc acagctgccg ggagaaaaga agaacggact gttcggaaac      720

ctgatcgcac tgagcctggg actgacaccg aacttcaaga gcaacttcga cctggcagaa      780

gacgcaaagc tgcagctgag caaggacaca tacgacgacg acctggacaa cctgctggca      840

cagatcggag accagtacgc agacctgttc ctggcagcaa agaacctgag cgacgcaatc      900

ctgctgagcg acatcctgag agtcaacaca gaaatcacaa aggcaccgct gagcgcaagc      960

atgatcaaga gatacgacga acaccaccag gacctgacac tgctgaaggc actggtcaga     1020

cagcagctgc cggaaaagta caaggaaatc ttcttcgacc agagcaagaa cggatacgca     1080

ggatacatcg acggaggagc aagccaggaa gaattctaca agttcatcaa gccgatcctg     1140

gaaaagatgg acggaacaga agaactgctg gtcaagctga acagagaaga cctgctgaga     1200

aagcagagaa cattcgacaa cggaagcatc ccgcaccaga tccacctggg agaactgcac     1260

gcaatcctga gaagacagga agacttctac ccgttcctga aggacaacag agaaaagatc     1320

gaaaagatcc tgacattcag aatcccgtac tacgtcggac cgctggcaag aggaaacagc     1380

agattcgcat ggatgacaag aaagagcgaa gaaacaatca caccgtggaa cttcgaagaa     1440

gtcgtcgaca agggagcaag cgcacagagc ttcatcgaaa gaatgacaaa cttcgacaag     1500

aacctgccga acgaaaaggt cctgccgaag cacagcctgc tgtacgaata cttcacagtc     1560

tacaacgaac tgacaaaggt caagtacgtc acagaaggaa tgagaaagcc ggcattcctg     1620

agcggagaac agaagaaggc aatcgtcgac ctgctgttca agacaaacag aaaggtcaca     1680

gtcaagcagc tgaaggaaga ctacttcaag aagatcgaat gcttcgacag cgtcgaaatc     1740

agcggagtcg aagacagatt caacgcaagc ctgggaacat accacgacct gctgaagatc     1800

atcaaggaca aggacttcct ggacaacgaa gaaaacgaag acatcctgga agacatcgtc     1860

ctgacactga cactgttcga agacagagaa atgatcgaag aaagactgaa gacatacgca     1920

cacctgttcg acgacaaggt catgaagcag ctgaagagaa gaagatacac aggatgggga     1980

agactgagca gaaagctgat caacggaatc agagacaagc agagcggaaa gacaatcctg     2040

gacttcctga agagcgacgg attcgcaaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacat tcaaggaaga catccagaag gcacaggtca gcggacaggg agacagcctg     2160

cacgaacaca tcgcaaacct ggcaggaagc ccggcaatca agaagggaat cctgcagaca     2220

gtcaaggtcg tcgacgaact ggtcaaggtc atgggaagac acaagccgga aaacatcgtc     2280

atcgaaatgg caagagaaaa ccagacaaca cagaagggac agaagaacag cagagaaaga     2340

atgaagagaa tcgaagaagg aatcaaggaa ctgggaagcc agatcctgaa ggaacacccg     2400

gtcgaaaaca cacagctgca gaacgaaaag ctgtacctgt actacctgca gaacggaaga     2460

gacatgtacg tcgaccagga actggacatc aacagactga gcgactacga cgtcgaccac     2520

atcgtcccgc agagcttcct gaaggacgac agcatcgaca acaaggtcct gacaagaagc     2580

gacaagaaca gaggaaagag cgacaacgtc ccgagcgaag aagtcgtcaa gaagatgaag     2640

aactactgga gacagctgct gaacgcaaag ctgatcacac agagaaagtt cgacaacctg     2700

acaaaggcag agagaggagg actgagcgaa ctggacaagg caggattcat caagagacag     2760

ctggtcgaaa caagacagat cacaaagcac gtcgcacaga tcctggacag cagaatgaac     2820

acaaagtacg acgaaaacga caagctgatc agagaagtca aggtcatcac actgaagagc     2880

aagctggtca gcgacttcag aaaggacttc cagttctaca aggtcagaga aatcaacaac     2940

taccaccacg cacacgacgc atacctgaac gcagtcgtcg gaacagcact gatcaagaag     3000

tacccgaagc tggaaagcga attcgtctac ggagactaca aggtctacga cgtcagaaag     3060

atgatcgcaa agagcgaaca ggaaatcgga aaggcaacag caaagtactt cttctacagc     3120

aacatcatga acttcttcaa gacagaaatc acactggcaa acggagaaat cagaaagaga     3180

ccgctgatcg aaacaaacgg agaaacagga gaaatcgtct gggacaaggg aagagacttc     3240

gcaacagtca gaaaggtcct gagcatgccg caggtcaaca tcgtcaagaa gacagaagtc     3300

cagacaggag gattcagcaa ggaaagcatc ctgccgaaga gaaacagcga caagctgatc     3360

gcaagaaaga aggactggga cccgaagaag tacggaggat tcgacagccc gacagtcgca     3420

tacagcgtcc tggtcgtcgc aaaggtcgaa aagggaaaga gcaagaagct gaagagcgtc     3480

aaggaactgc tgggaatcac aatcatggaa agaagcagct tcgaaaagaa cccgatcgac     3540

ttcctggaag caaagggata caaggaagtc aagaaggacc tgatcatcaa gctgccgaag     3600

tacagcctgt tcgaactgga aaacggaaga aagagaatgc tggcaagcgc aggagaactg     3660

cagaagggaa acgaactggc actgccgagc aagtacgtca acttcctgta cctggcaagc     3720

cactacgaaa agctgaaggg aagcccggaa gacaacgaac agaagcagct gttcgtcgaa     3780

cagcacaagc actacctgga cgaaatcatc gaacagatca gcgaattcag caagagagtc     3840

atcctggcag acgcaaacct ggacaaggtc ctgagcgcat acaacaagca cagagacaag     3900

ccgatcagag aacaggcaga aaacatcatc cacctgttca cactgacaaa cctgggagca     3960

ccggcagcat tcaagtactt cgacacaaca atcgacagaa agagatacac aagcacaaag     4020

gaagtcctgg acgcaacact gatccaccag agcatcacag gactgtacga aacaagaatc     4080

gacctgagcc agctgggagg agacggagga ggaagcagag cagcaaagag aaagtacttc     4140

gcagtctag                                                             4149


<210>  141
<211>  4128
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using codons with generally high expression 
       in humans (no start or stop codons; suitable for inclusion in 
       fusion protein coding sequence)

<400>  141
cctaagaaaa agcggaaggt cgacggggat aagaagtact caatcgggct ggatatcgga       60

actaattccg tgggttgggc agtgatcacg gatgaataca aagtgccgtc caagaagttc      120

aaggtcctgg ggaacaccga tagacacagc atcaagaaaa atctcatcgg agccctgctg      180

tttgactccg gcgaaaccgc agaagcgacc cggctcaaac gtaccgcgag gcgacgctac      240

acccggcgga agaatcgcat ctgctatctg caagagatct tttcgaacga aatggcaaag      300

gtcgacgaca gcttcttcca ccgcctggaa gaatctttcc tggtggagga ggacaagaag      360

catgaacggc atcctatctt tggaaacatc gtcgacgaag tggcgtacca cgaaaagtac      420

ccgaccatct accatctgcg gaagaagttg gttgactcaa ctgacaaggc cgacctcaga      480

ttgatctact tggccctcgc ccatatgatc aaattccgcg gacacttcct gatcgaaggc      540

gatctgaacc ctgataactc cgacgtggat aagcttttca ttcaactggt gcagacctac      600

aaccaactgt tcgaagaaaa cccaatcaat gctagcggcg tcgatgccaa ggccatcctg      660

tccgcccggc tgtcgaagtc gcggcgcctc gaaaacctga tcgcacagct gccgggagag      720

aaaaagaacg gacttttcgg caacttgatc gctctctcac tgggactcac tcccaatttc      780

aagtccaatt ttgacctggc cgaggacgcg aagctgcaac tctcaaagga cacctacgac      840

gacgacttgg acaatttgct ggcacaaatt ggcgatcagt acgcggatct gttccttgcc      900

gctaagaacc tttcggacgc aatcttgctg tccgatatcc tgcgcgtgaa caccgaaata      960

accaaagcgc cgcttagcgc ctcgatgatt aagcggtacg acgagcatca ccaggatctc     1020

acgctgctca aagcgctcgt gagacagcaa ctgcctgaaa agtacaagga gatcttcttc     1080

gaccagtcca agaatgggta cgcagggtac atcgatggag gcgctagcca ggaagagttc     1140

tataagttca tcaagccaat cctggaaaag atggacggaa ccgaagaact gctggtcaag     1200

ctgaacaggg aggatctgct ccggaaacag agaacctttg acaacggatc cattccccac     1260

cagatccatc tgggtgagct gcacgccatc ttgcggcgcc aggaggactt ttacccattc     1320

ctcaaggaca accgggaaaa gatcgagaaa attctgacgt tccgcatccc gtattacgtg     1380

ggcccactgg cgcgcggcaa ttcgcgcttc gcgtggatga ctagaaaatc agaggaaacc     1440

atcactcctt ggaatttcga ggaagttgtg gataagggag cttcggcaca aagcttcatc     1500

gaacgaatga ccaacttcga caagaatctc ccaaacgaga aggtgcttcc taagcacagc     1560

ctcctttacg aatacttcac tgtctacaac gaactgacta aagtgaaata cgttactgaa     1620

ggaatgagga agccggcctt tctgtccgga gaacagaaga aagcaattgt cgatctgctg     1680

ttcaagacca accgcaaggt gaccgtcaag cagcttaaag aggactactt caagaagatc     1740

gagtgtttcg actcagtgga aatcagcggg gtggaggaca gattcaacgc ttcgctggga     1800

acctatcatg atctcctgaa gatcatcaag gacaaggact tccttgacaa cgaggagaac     1860

gaggacatcc tggaagatat cgtcctgacc ttgacccttt tcgaggatcg cgagatgatc     1920

gaggagaggc ttaagaccta cgctcatctc ttcgacgata aggtcatgaa acaactcaag     1980

cgccgccggt acactggttg gggccgcctc tcccgcaagc tgatcaacgg tattcgcgat     2040

aaacagagcg gtaaaactat cctggatttc ctcaaatcgg atggcttcgc taatcgtaac     2100

ttcatgcaat tgatccacga cgacagcctg acctttaagg aggacatcca aaaagcacaa     2160

gtgtccggac agggagactc actccatgaa cacatcgcga atctggccgg ttcgccggcg     2220

attaagaagg gaattctgca aactgtgaag gtggtcgacg agctggtgaa ggtcatggga     2280

cggcacaaac cggagaatat cgtgattgaa atggcccgag aaaaccagac tacccagaag     2340

ggccagaaaa actcccgcga aaggatgaag cggatcgaag aaggaatcaa ggagctgggc     2400

agccagatcc tgaaagagca cccggtggaa aacacgcagc tgcagaacga gaagctctac     2460

ctgtactatt tgcaaaatgg acgggacatg tacgtggacc aagagctgga catcaatcgg     2520

ttgtctgatt acgacgtgga ccacatcgtt ccacagtcct ttctgaagga tgactcgatc     2580

gataacaagg tgttgactcg cagcgacaag aacagaggga agtcagataa tgtgccatcg     2640

gaggaggtcg tgaagaagat gaagaattac tggcggcagc tcctgaatgc gaagctgatt     2700

acccagagaa agtttgacaa tctcactaaa gccgagcgcg gcggactctc agagctggat     2760

aaggctggat tcatcaaacg gcagctggtc gagactcggc agattaccaa gcacgtggcg     2820

cagatcttgg actcccgcat gaacactaaa tacgacgaga acgataagct catccgggaa     2880

gtgaaggtga ttaccctgaa aagcaaactt gtgtcggact ttcggaagga ctttcagttt     2940

tacaaagtga gagaaatcaa caactaccat cacgcgcatg acgcatacct caacgctgtg     3000

gtcggtaccg ccctgatcaa aaagtaccct aaacttgaat cggagtttgt gtacggagac     3060

tacaaggtct acgacgtgag gaagatgata gccaagtccg aacaggaaat cgggaaagca     3120

actgcgaaat acttctttta ctcaaacatc atgaactttt tcaagactga aattacgctg     3180

gccaatggag aaatcaggaa gaggccactg atcgaaacta acggagaaac gggcgaaatc     3240

gtgtgggaca agggcaggga cttcgcaact gttcgcaaag tgctctctat gccgcaagtc     3300

aatattgtga agaaaaccga agtgcaaacc ggcggatttt caaaggaatc gatcctccca     3360

aagagaaata gcgacaagct cattgcacgc aagaaagact gggacccgaa gaagtacgga     3420

ggattcgatt cgccgactgt cgcatactcc gtcctcgtgg tggccaaggt ggagaaggga     3480

aagagcaaaa agctcaaatc cgtcaaagag ctgctgggga ttaccatcat ggaacgatcc     3540

tcgttcgaga agaacccgat tgatttcctc gaggcgaagg gttacaagga ggtgaagaag     3600

gatctgatca tcaaactccc caagtactca ctgttcgaac tggaaaatgg tcggaagcgc     3660

atgctggctt cggccggaga actccaaaaa ggaaatgagc tggccttgcc tagcaagtac     3720

gtcaacttcc tctatcttgc ttcgcactac gaaaaactca aagggtcacc ggaagataac     3780

gaacagaagc agcttttcgt ggagcagcac aagcattatc tggatgaaat catcgaacaa     3840

atctccgagt tttcaaagcg cgtgatcctc gccgacgcca acctcgacaa agtcctgtcg     3900

gcctacaata agcatagaga taagccgatc agagaacagg ccgagaacat tatccacttg     3960

ttcaccctga ctaacctggg agccccagcc gccttcaagt acttcgatac tactatcgat     4020

cgcaaaagat acacgtccac caaggaagtt ctggacgcga ccctgatcca ccaaagcatc     4080

actggactct acgaaactag gatcgatctg tcgcagctgg gtggcgat                  4128


<210>  142
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using long half life codons of Table 4 (no 
       start or stop codons; suitable for inclusion in fusion protein 
       coding sequence)

<400>  142
gacaagaagt actctatcgg tttggacatc ggtaccaact ctgtcggttg ggccgtcatc       60

accgacgaat acaaggtccc atctaagaag ttcaaggtct tgggtaacac cgacagacac      120

tctatcaaga agaacttgat cggtgccttg ttgttcgact ctggtgaaac cgccgaagcc      180

accagattga agagaaccgc cagaagaaga tacaccagaa gaaagaacag aatctgctac      240

ttgcaagaaa tcttctctaa cgaaatggcc aaggtcgacg actctttctt ccacagattg      300

gaagaatctt tcttggtcga agaagacaag aagcacgaaa gacacccaat cttcggtaac      360

atcgtcgacg aagtcgccta ccacgaaaag tacccaacca tctaccactt gagaaagaag      420

ttggtcgact ctaccgacaa ggccgacttg agattgatct acttggcctt ggcccacatg      480

atcaagttca gaggtcactt cttgatcgaa ggtgacttga acccagacaa ctctgacgtc      540

gacaagttgt tcatccaatt ggtccaaacc tacaaccaat tgttcgaaga aaacccaatc      600

aacgcctctg gtgtcgacgc caaggccatc ttgtctgcca gattgtctaa gagcagaaga      660

ttggaaaact tgatcgccca attgccaggt gaaaagaaga acggtttgtt cggtaacttg      720

atcgccttgt ctttgggttt gaccccaaac ttcaagtcta acttcgactt ggccgaagac      780

gccaagttgc aattgtctaa ggacacctac gacgacgact tggacaactt gttggcccaa      840

atcggtgacc aatacgccga cttgttcttg gccgccaaga acttgtctga cgccatcttg      900

ttgtctgaca tcttgagagt caacaccgaa atcaccaagg ccccattgtc tgcctctatg      960

atcaagagat acgacgaaca ccaccaagac ttgaccttgt tgaaggcctt ggtcagacaa     1020

caattgccag aaaagtacaa ggaaatcttc ttcgaccaat ctaagaacgg ttacgccggt     1080

tacatcgacg gtggtgcctc tcaagaagaa ttctacaagt tcatcaagcc aatcttggaa     1140

aagatggacg gtaccgaaga attgttggtc aagttgaaca gagaagactt gttgagaaag     1200

caaagaacct tcgacaacgg ttctatccca caccaaatcc acttgggtga attgcacgcc     1260

atcttgagaa gacaagaaga cttctaccca ttcttgaagg acaacagaga aaagatcgaa     1320

aagatcttga ccttcagaat cccatactac gtcggtccat tggccagagg taacagcaga     1380

ttcgcctgga tgaccagaaa gtctgaagaa accatcaccc catggaactt cgaagaagtc     1440

gtcgacaagg gtgcctctgc ccaatctttc atcgaaagaa tgaccaactt cgacaagaac     1500

ttgccaaacg aaaaggtctt gccaaagcac tctttgttgt acgaatactt caccgtctac     1560

aacgaattga ccaaggtcaa gtacgtcacc gaaggtatga gaaagccagc cttcttgtct     1620

ggtgaacaaa agaaggccat cgtcgacttg ttgttcaaga ccaacagaaa ggtcaccgtc     1680

aagcaattga aggaagacta cttcaagaag atcgaatgct tcgactctgt cgaaatctct     1740

ggtgtcgaag acagattcaa cgcctctttg ggtacctacc acgacttgtt gaagatcatc     1800

aaggacaagg acttcttgga caacgaagaa aacgaagaca tcttggaaga catcgtcttg     1860

accttgacct tgttcgaaga cagagaaatg atcgaagaaa gattgaagac ctacgcccac     1920

ttgttcgacg acaaggtcat gaagcaattg aagagaagaa gatacaccgg ttggggtaga     1980

ttgagcagaa agttgatcaa cggtatcaga gacaagcaat ctggtaagac catcttggac     2040

ttcttgaagt ctgacggttt cgccaacaga aacttcatgc aattgatcca cgacgactct     2100

ttgaccttca aggaagacat ccaaaaggcc caagtctctg gtcaaggtga ctctttgcac     2160

gaacacatcg ccaacttggc cggttctcca gccatcaaga agggtatctt gcaaaccgtc     2220

aaggtcgtcg acgaattggt caaggtcatg ggtagacaca agccagaaaa catcgtcatc     2280

gaaatggcca gagaaaacca aaccacccaa aagggtcaaa agaacagcag agaaagaatg     2340

aagagaatcg aagaaggtat caaggaattg ggttctcaaa tcttgaagga acacccagtc     2400

gaaaacaccc aattgcaaaa cgaaaagttg tacttgtact acttgcaaaa cggtagagac     2460

atgtacgtcg accaagaatt ggacatcaac agattgtctg actacgacgt cgaccacatc     2520

gtcccacaat ctttcttgaa ggacgactct atcgacaaca aggtcttgac cagatctgac     2580

aagaacagag gtaagtctga caacgtccca tctgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac aattgttgaa cgccaagttg atcacccaaa gaaagttcga caacttgacc     2700

aaggccgaaa gaggtggttt gtctgaattg gacaaggccg gtttcatcaa gagacaattg     2760

gtcgaaacca gacaaatcac caagcacgtc gcccaaatct tggacagcag aatgaacacc     2820

aagtacgacg aaaacgacaa gttgatcaga gaagtcaagg tcatcacctt gaagtctaag     2880

ttggtctctg acttcagaaa ggacttccaa ttctacaagg tcagagaaat caacaactac     2940

caccacgccc acgacgccta cttgaacgcc gtcgtcggta ccgccttgat caagaagtac     3000

ccaaagttgg aatctgaatt cgtctacggt gactacaagg tctacgacgt cagaaagatg     3060

atcgccaagt ctgaacaaga aatcggtaag gccaccgcca agtacttctt ctactctaac     3120

atcatgaact tcttcaagac cgaaatcacc ttggccaacg gtgaaatcag aaagagacca     3180

ttgatcgaaa ccaacggtga aaccggtgaa atcgtctggg acaagggtag agacttcgcc     3240

accgtcagaa aggtcttgtc tatgccacaa gtcaacatcg tcaagaagac cgaagtccaa     3300

accggtggtt tctctaagga atctatcttg ccaaagagaa actctgacaa gttgatcgcc     3360

agaaagaagg actgggaccc aaagaagtac ggtggtttcg actctccaac cgtcgcctac     3420

tctgtcttgg tcgtcgccaa ggtcgaaaag ggtaagtcta agaagttgaa gtctgtcaag     3480

gaattgttgg gtatcaccat catggaaaga tcttctttcg aaaagaaccc aatcgacttc     3540

ttggaagcca agggttacaa ggaagtcaag aaggacttga tcatcaagtt gccaaagtac     3600

tctttgttcg aattggaaaa cggtagaaag agaatgttgg cctctgccgg tgaattgcaa     3660

aagggtaacg aattggcctt gccatctaag tacgtcaact tcttgtactt ggcctctcac     3720

tacgaaaagt tgaagggttc tccagaagac aacgaacaaa agcaattgtt cgtcgaacaa     3780

cacaagcact acttggacga aatcatcgaa caaatctctg aattctctaa gagagtcatc     3840

ttggccgacg ccaacttgga caaggtcttg tctgcctaca acaagcacag agacaagcca     3900

atcagagaac aagccgaaaa catcatccac ttgttcacct tgaccaactt gggtgcccca     3960

gccgccttca agtacttcga caccaccatc gacagaaaga gatacacctc taccaaggaa     4020

gtcttggacg ccaccttgat ccaccaatct atcaccggtt tgtacgaaac cagaatcgac     4080

ttgtctcaat tgggtggtga cggtggtggt tctccaaaga agaagagaaa ggtc           4134


<210>  143
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using U rich codons of Table 4 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  143
gataaaaaat attctattgg tttagatatt ggtactaatt ctgttggttg ggctgttatt       60

actgatgaat ataaagttcc ttctaaaaaa tttaaagttt taggtaatac tgatcgtcat      120

tctattaaaa aaaatttaat tggtgcttta ttatttgatt ctggtgaaac tgctgaagct      180

actcgtttaa aacgtactgc tcgtcgtcgt tatactcgtc gtaaaaatcg tatttgttat      240

ttacaagaaa ttttttctaa tgaaatggct aaagttgatg attctttttt tcatcgttta      300

gaagaatctt ttttagttga agaagataaa aaacatgaac gtcatcctat ttttggtaat      360

attgttgatg aagttgctta tcatgaaaaa tatcctacta tttatcattt acgtaaaaaa      420

ttagttgatt ctactgataa agctgattta cgtttaattt atttagcttt agctcatatg      480

attaaatttc gtggtcattt tttaattgaa ggtgatttaa atcctgataa ttctgatgtt      540

gataaattat ttattcaatt agttcaaact tataatcaat tatttgaaga aaatcctatt      600

aatgcttctg gtgttgatgc taaagctatt ttatctgctc gtttatctaa atctcgtcgt      660

ttagaaaatt taattgctca attacctggt gaaaaaaaaa atggtttatt tggtaattta      720

attgctttat ctttaggttt aactcctaat tttaaatcta attttgattt agctgaagat      780

gctaaattac aattatctaa agatacttat gatgatgatt tagataattt attagctcaa      840

attggtgatc aatatgctga tttattttta gctgctaaaa atttatctga tgctatttta      900

ttatctgata ttttacgtgt taatactgaa attactaaag ctcctttatc tgcttctatg      960

attaaacgtt atgatgaaca tcatcaagat ttaactttat taaaagcttt agttcgtcaa     1020

caattacctg aaaaatataa agaaattttt tttgatcaat ctaaaaatgg ttatgctggt     1080

tatattgatg gtggtgcttc tcaagaagaa ttttataaat ttattaaacc tattttagaa     1140

aaaatggatg gtactgaaga attattagtt aaattaaatc gtgaagattt attacgtaaa     1200

caacgtactt ttgataatgg ttctattcct catcaaattc atttaggtga attacatgct     1260

attttacgtc gtcaagaaga tttttatcct tttttaaaag ataatcgtga aaaaattgaa     1320

aaaattttaa cttttcgtat tccttattat gttggtcctt tagctcgtgg taattctcgt     1380

tttgcttgga tgactcgtaa atctgaagaa actattactc cttggaattt tgaagaagtt     1440

gttgataaag gtgcttctgc tcaatctttt attgaacgta tgactaattt tgataaaaat     1500

ttacctaatg aaaaagtttt acctaaacat tctttattat atgaatattt tactgtttat     1560

aatgaattaa ctaaagttaa atatgttact gaaggtatgc gtaaacctgc ttttttatct     1620

ggtgaacaaa aaaaagctat tgttgattta ttatttaaaa ctaatcgtaa agttactgtt     1680

aaacaattaa aagaagatta ttttaaaaaa attgaatgtt ttgattctgt tgaaatttct     1740

ggtgttgaag atcgttttaa tgcttcttta ggtacttatc atgatttatt aaaaattatt     1800

aaagataaag attttttaga taatgaagaa aatgaagata ttttagaaga tattgtttta     1860

actttaactt tatttgaaga tcgtgaaatg attgaagaac gtttaaaaac ttatgctcat     1920

ttatttgatg ataaagttat gaaacaatta aaacgtcgtc gttatactgg ttggggtcgt     1980

ttatctcgta aattaattaa tggtattcgt gataaacaat ctggtaaaac tattttagat     2040

tttttaaaat ctgatggttt tgctaatcgt aattttatgc aattaattca tgatgattct     2100

ttaactttta aagaagatat tcaaaaagct caagtttctg gtcaaggtga ttctttacat     2160

gaacatattg ctaatttagc tggttctcct gctattaaaa aaggtatttt acaaactgtt     2220

aaagttgttg atgaattagt taaagttatg ggtcgtcata aacctgaaaa tattgttatt     2280

gaaatggctc gtgaaaatca aactactcaa aaaggtcaaa aaaattctcg tgaacgtatg     2340

aaacgtattg aagaaggtat taaagaatta ggttctcaaa ttttaaaaga acatcctgtt     2400

gaaaatactc aattacaaaa tgaaaaatta tatttatatt atttacaaaa tggtcgtgat     2460

atgtatgttg atcaagaatt agatattaat cgtttatctg attatgatgt tgatcatatt     2520

gttcctcaat cttttttaaa agatgattct attgataata aagttttaac tcgttctgat     2580

aaaaatcgtg gtaaatctga taatgttcct tctgaagaag ttgttaaaaa aatgaaaaat     2640

tattggcgtc aattattaaa tgctaaatta attactcaac gtaaatttga taatttaact     2700

aaagctgaac gtggtggttt atctgaatta gataaagctg gttttattaa acgtcaatta     2760

gttgaaactc gtcaaattac taaacatgtt gctcaaattt tagattctcg tatgaatact     2820

aaatatgatg aaaatgataa attaattcgt gaagttaaag ttattacttt aaaatctaaa     2880

ttagtttctg attttcgtaa agattttcaa ttttataaag ttcgtgaaat taataattat     2940

catcatgctc atgatgctta tttaaatgct gttgttggta ctgctttaat taaaaaatat     3000

cctaaattag aatctgaatt tgtttatggt gattataaag tttatgatgt tcgtaaaatg     3060

attgctaaat ctgaacaaga aattggtaaa gctactgcta aatatttttt ttattctaat     3120

attatgaatt tttttaaaac tgaaattact ttagctaatg gtgaaattcg taaacgtcct     3180

ttaattgaaa ctaatggtga aactggtgaa attgtttggg ataaaggtcg tgattttgct     3240

actgttcgta aagttttatc tatgcctcaa gttaatattg ttaaaaaaac tgaagttcaa     3300

actggtggtt tttctaaaga atctatttta cctaaacgta attctgataa attaattgct     3360

cgtaaaaaag attgggatcc taaaaaatat ggtggttttg attctcctac tgttgcttat     3420

tctgttttag ttgttgctaa agttgaaaaa ggtaaatcta aaaaattaaa atctgttaaa     3480

gaattattag gtattactat tatggaacgt tcttcttttg aaaaaaatcc tattgatttt     3540

ttagaagcta aaggttataa agaagttaaa aaagatttaa ttattaaatt acctaaatat     3600

tctttatttg aattagaaaa tggtcgtaaa cgtatgttag cttctgctgg tgaattacaa     3660

aaaggtaatg aattagcttt accttctaaa tatgttaatt ttttatattt agcttctcat     3720

tatgaaaaat taaaaggttc tcctgaagat aatgaacaaa aacaattatt tgttgaacaa     3780

cataaacatt atttagatga aattattgaa caaatttctg aattttctaa acgtgttatt     3840

ttagctgatg ctaatttaga taaagtttta tctgcttata ataaacatcg tgataaacct     3900

attcgtgaac aagctgaaaa tattattcat ttatttactt taactaattt aggtgctcct     3960

gctgctttta aatattttga tactactatt gatcgtaaac gttatacttc tactaaagaa     4020

gttttagatg ctactttaat tcatcaatct attactggtt tatatgaaac tcgtattgat     4080

ttatctcaat taggtggtga tggtggtggt tctcctaaaa aaaaacgtaa agtt           4134


<210>  144
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low G codons of Table 4 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  144
gacaaaaaat actccatcgg cctcgacatc ggcaccaact ccgtcggctg ggccgtcatc       60

accgacgaat acaaagtccc ctccaaaaaa ttcaaagtcc tcggcaacac cgacagacac      120

tccatcaaaa aaaacctcat cggcgccctc ctcttcgact ccggcgaaac cgccgaagcc      180

accagactca aaagaaccgc cagaagaaga tacaccagaa gaaaaaacag aatctgctac      240

ctccaagaaa tcttctccaa cgaaatggcc aaagtcgacg actccttctt ccacagactc      300

gaagaatcct tcctcgtcga agaagacaaa aaacacgaaa gacaccccat cttcggcaac      360

atcgtcgacg aagtcgccta ccacgaaaaa taccccacca tctaccacct cagaaaaaaa      420

ctcgtcgact ccaccgacaa agccgacctc agactcatct acctcgccct cgcccacatg      480

atcaaattca gaggccactt cctcatcgaa ggcgacctca accccgacaa ctccgacgtc      540

gacaaactct tcatccaact cgtccaaacc tacaaccaac tcttcgaaga aaaccccatc      600

aacgcctccg gcgtcgacgc caaagccatc ctctccgcca gactctccaa atccagaaga      660

ctcgaaaacc tcatcgccca actccccggc gaaaaaaaaa acggcctctt cggcaacctc      720

atcgccctct ccctcggcct cacccccaac ttcaaatcca acttcgacct cgccgaagac      780

gccaaactcc aactctccaa agacacctac gacgacgacc tcgacaacct cctcgcccaa      840

atcggcgacc aatacgccga cctcttcctc gccgccaaaa acctctccga cgccatcctc      900

ctctccgaca tcctcagagt caacaccgaa atcaccaaag cccccctctc cgcctccatg      960

atcaaaagat acgacgaaca ccaccaagac ctcaccctcc tcaaagccct cgtcagacaa     1020

caactccccg aaaaatacaa agaaatcttc ttcgaccaat ccaaaaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaagaagaa ttctacaaat tcatcaaacc catcctcgaa     1140

aaaatggacg gcaccgaaga actcctcgtc aaactcaaca gagaagacct cctcagaaaa     1200

caaagaacct tcgacaacgg ctccatcccc caccaaatcc acctcggcga actccacgcc     1260

atcctcagaa gacaagaaga cttctacccc ttcctcaaag acaacagaga aaaaatcgaa     1320

aaaatcctca ccttcagaat cccctactac gtcggccccc tcgccagagg caactccaga     1380

ttcgcctgga tgaccagaaa atccgaagaa accatcaccc cctggaactt cgaagaagtc     1440

gtcgacaaag gcgcctccgc ccaatccttc atcgaaagaa tgaccaactt cgacaaaaac     1500

ctccccaacg aaaaagtcct ccccaaacac tccctcctct acgaatactt caccgtctac     1560

aacgaactca ccaaagtcaa atacgtcacc gaaggcatga gaaaacccgc cttcctctcc     1620

ggcgaacaaa aaaaagccat cgtcgacctc ctcttcaaaa ccaacagaaa agtcaccgtc     1680

aaacaactca aagaagacta cttcaaaaaa atcgaatgct tcgactccgt cgaaatctcc     1740

ggcgtcgaag acagattcaa cgcctccctc ggcacctacc acgacctcct caaaatcatc     1800

aaagacaaag acttcctcga caacgaagaa aacgaagaca tcctcgaaga catcgtcctc     1860

accctcaccc tcttcgaaga cagagaaatg atcgaagaaa gactcaaaac ctacgcccac     1920

ctcttcgacg acaaagtcat gaaacaactc aaaagaagaa gatacaccgg ctggggcaga     1980

ctctccagaa aactcatcaa cggcatcaga gacaaacaat ccggcaaaac catcctcgac     2040

ttcctcaaat ccgacggctt cgccaacaga aacttcatgc aactcatcca cgacgactcc     2100

ctcaccttca aagaagacat ccaaaaagcc caagtctccg gccaaggcga ctccctccac     2160

gaacacatcg ccaacctcgc cggctccccc gccatcaaaa aaggcatcct ccaaaccgtc     2220

aaagtcgtcg acgaactcgt caaagtcatg ggcagacaca aacccgaaaa catcgtcatc     2280

gaaatggcca gagaaaacca aaccacccaa aaaggccaaa aaaactccag agaaagaatg     2340

aaaagaatcg aagaaggcat caaagaactc ggctcccaaa tcctcaaaga acaccccgtc     2400

gaaaacaccc aactccaaaa cgaaaaactc tacctctact acctccaaaa cggcagagac     2460

atgtacgtcg accaagaact cgacatcaac agactctccg actacgacgt cgaccacatc     2520

gtcccccaat ccttcctcaa agacgactcc atcgacaaca aagtcctcac cagatccgac     2580

aaaaacagag gcaaatccga caacgtcccc tccgaagaag tcgtcaaaaa aatgaaaaac     2640

tactggagac aactcctcaa cgccaaactc atcacccaaa gaaaattcga caacctcacc     2700

aaagccgaaa gaggcggcct ctccgaactc gacaaagccg gcttcatcaa aagacaactc     2760

gtcgaaacca gacaaatcac caaacacgtc gcccaaatcc tcgactccag aatgaacacc     2820

aaatacgacg aaaacgacaa actcatcaga gaagtcaaag tcatcaccct caaatccaaa     2880

ctcgtctccg acttcagaaa agacttccaa ttctacaaag tcagagaaat caacaactac     2940

caccacgccc acgacgccta cctcaacgcc gtcgtcggca ccgccctcat caaaaaatac     3000

cccaaactcg aatccgaatt cgtctacggc gactacaaag tctacgacgt cagaaaaatg     3060

atcgccaaat ccgaacaaga aatcggcaaa gccaccgcca aatacttctt ctactccaac     3120

atcatgaact tcttcaaaac cgaaatcacc ctcgccaacg gcgaaatcag aaaaagaccc     3180

ctcatcgaaa ccaacggcga aaccggcgaa atcgtctggg acaaaggcag agacttcgcc     3240

accgtcagaa aagtcctctc catgccccaa gtcaacatcg tcaaaaaaac cgaagtccaa     3300

accggcggct tctccaaaga atccatcctc cccaaaagaa actccgacaa actcatcgcc     3360

agaaaaaaag actgggaccc caaaaaatac ggcggcttcg actcccccac cgtcgcctac     3420

tccgtcctcg tcgtcgccaa agtcgaaaaa ggcaaatcca aaaaactcaa atccgtcaaa     3480

gaactcctcg gcatcaccat catggaaaga tcctccttcg aaaaaaaccc catcgacttc     3540

ctcgaagcca aaggctacaa agaagtcaaa aaagacctca tcatcaaact ccccaaatac     3600

tccctcttcg aactcgaaaa cggcagaaaa agaatgctcg cctccgccgg cgaactccaa     3660

aaaggcaacg aactcgccct cccctccaaa tacgtcaact tcctctacct cgcctcccac     3720

tacgaaaaac tcaaaggctc ccccgaagac aacgaacaaa aacaactctt cgtcgaacaa     3780

cacaaacact acctcgacga aatcatcgaa caaatctccg aattctccaa aagagtcatc     3840

ctcgccgacg ccaacctcga caaagtcctc tccgcctaca acaaacacag agacaaaccc     3900

atcagagaac aagccgaaaa catcatccac ctcttcaccc tcaccaacct cggcgccccc     3960

gccgccttca aatacttcga caccaccatc gacagaaaaa gatacacctc caccaaagaa     4020

gtcctcgacg ccaccctcat ccaccaatcc atcaccggcc tctacgaaac cagaatcgac     4080

ctctcccaac tcggcggcga cggcggcggc tcccccaaaa aaaaaagaaa agtc           4134


<210>  145
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low C codons of Table 4 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  145
gataagaagt atagtattgg attggatatt ggaacaaata gtgtgggatg ggctgtgatt       60

acagatgagt ataaggtgcc tagtaagaag tttaaggtgt tgggaaatac agatagacat      120

agtattaaga agaatttgat tggagctttg ttgtttgata gtggagagac agctgaggct      180

acaagattga agagaacagc tagaagaaga tatacaagaa gaaagaatag aatttgttat      240

ttgcaggaga tttttagtaa tgagatggct aaggtggatg atagtttttt tcatagattg      300

gaggagagtt ttttggtgga ggaggataag aagcatgaga gacatcctat ttttggaaat      360

attgtggatg aggtggctta tcatgagaag tatcctacaa tttatcattt gagaaagaag      420

ttggtggata gtacagataa ggctgatttg agattgattt atttggcttt ggctcatatg      480

attaagttta gaggacattt tttgattgag ggagatttga atcctgataa tagtgatgtg      540

gataagttgt ttattcagtt ggtgcagaca tataatcagt tgtttgagga gaatcctatt      600

aatgctagtg gagtggatgc taaggctatt ttgagtgcta gattgagtaa gagtagaaga      660

ttggagaatt tgattgctca gttgcctgga gagaagaaga atggattgtt tggaaatttg      720

attgctttga gtttgggatt gacacctaat tttaagagta attttgattt ggctgaggat      780

gctaagttgc agttgagtaa ggatacatat gatgatgatt tggataattt gttggctcag      840

attggagatc agtatgctga tttgtttttg gctgctaaga atttgagtga tgctattttg      900

ttgagtgata ttttgagagt gaatacagag attacaaagg ctcctttgag tgctagtatg      960

attaagagat atgatgagca tcatcaggat ttgacattgt tgaaggcttt ggtgagacag     1020

cagttgcctg agaagtataa ggagattttt tttgatcaga gtaagaatgg atatgctgga     1080

tatattgatg gaggagctag tcaggaggag ttttataagt ttattaagcc tattttggag     1140

aagatggatg gaacagagga gttgttggtg aagttgaata gagaggattt gttgagaaag     1200

cagagaacat ttgataatgg aagtattcct catcagattc atttgggaga gttgcatgct     1260

attttgagaa gacaggagga tttttatcct tttttgaagg ataatagaga gaagattgag     1320

aagattttga catttagaat tccttattat gtgggacctt tggctagagg aaatagtaga     1380

tttgcttgga tgacaagaaa gagtgaggag acaattacac cttggaattt tgaggaggtg     1440

gtggataagg gagctagtgc tcagagtttt attgagagaa tgacaaattt tgataagaat     1500

ttgcctaatg agaaggtgtt gcctaagcat agtttgttgt atgagtattt tacagtgtat     1560

aatgagttga caaaggtgaa gtatgtgaca gagggaatga gaaagcctgc ttttttgagt     1620

ggagagcaga agaaggctat tgtggatttg ttgtttaaga caaatagaaa ggtgacagtg     1680

aagcagttga aggaggatta ttttaagaag attgagtgtt ttgatagtgt ggagattagt     1740

ggagtggagg atagatttaa tgctagtttg ggaacatatc atgatttgtt gaagattatt     1800

aaggataagg attttttgga taatgaggag aatgaggata ttttggagga tattgtgttg     1860

acattgacat tgtttgagga tagagagatg attgaggaga gattgaagac atatgctcat     1920

ttgtttgatg ataaggtgat gaagcagttg aagagaagaa gatatacagg atggggaaga     1980

ttgagtagaa agttgattaa tggaattaga gataagcaga gtggaaagac aattttggat     2040

tttttgaaga gtgatggatt tgctaataga aattttatgc agttgattca tgatgatagt     2100

ttgacattta aggaggatat tcagaaggct caggtgagtg gacagggaga tagtttgcat     2160

gagcatattg ctaatttggc tggaagtcct gctattaaga agggaatttt gcagacagtg     2220

aaggtggtgg atgagttggt gaaggtgatg ggaagacata agcctgagaa tattgtgatt     2280

gagatggcta gagagaatca gacaacacag aagggacaga agaatagtag agagagaatg     2340

aagagaattg aggagggaat taaggagttg ggaagtcaga ttttgaagga gcatcctgtg     2400

gagaatacac agttgcagaa tgagaagttg tatttgtatt atttgcagaa tggaagagat     2460

atgtatgtgg atcaggagtt ggatattaat agattgagtg attatgatgt ggatcatatt     2520

gtgcctcaga gttttttgaa ggatgatagt attgataata aggtgttgac aagaagtgat     2580

aagaatagag gaaagagtga taatgtgcct agtgaggagg tggtgaagaa gatgaagaat     2640

tattggagac agttgttgaa tgctaagttg attacacaga gaaagtttga taatttgaca     2700

aaggctgaga gaggaggatt gagtgagttg gataaggctg gatttattaa gagacagttg     2760

gtggagacaa gacagattac aaagcatgtg gctcagattt tggatagtag aatgaataca     2820

aagtatgatg agaatgataa gttgattaga gaggtgaagg tgattacatt gaagagtaag     2880

ttggtgagtg attttagaaa ggattttcag ttttataagg tgagagagat taataattat     2940

catcatgctc atgatgctta tttgaatgct gtggtgggaa cagctttgat taagaagtat     3000

cctaagttgg agagtgagtt tgtgtatgga gattataagg tgtatgatgt gagaaagatg     3060

attgctaaga gtgagcagga gattggaaag gctacagcta agtatttttt ttatagtaat     3120

attatgaatt tttttaagac agagattaca ttggctaatg gagagattag aaagagacct     3180

ttgattgaga caaatggaga gacaggagag attgtgtggg ataagggaag agattttgct     3240

acagtgagaa aggtgttgag tatgcctcag gtgaatattg tgaagaagac agaggtgcag     3300

acaggaggat ttagtaagga gagtattttg cctaagagaa atagtgataa gttgattgct     3360

agaaagaagg attgggatcc taagaagtat ggaggatttg atagtcctac agtggcttat     3420

agtgtgttgg tggtggctaa ggtggagaag ggaaagagta agaagttgaa gagtgtgaag     3480

gagttgttgg gaattacaat tatggagaga agtagttttg agaagaatcc tattgatttt     3540

ttggaggcta agggatataa ggaggtgaag aaggatttga ttattaagtt gcctaagtat     3600

agtttgtttg agttggagaa tggaagaaag agaatgttgg ctagtgctgg agagttgcag     3660

aagggaaatg agttggcttt gcctagtaag tatgtgaatt ttttgtattt ggctagtcat     3720

tatgagaagt tgaagggaag tcctgaggat aatgagcaga agcagttgtt tgtggagcag     3780

cataagcatt atttggatga gattattgag cagattagtg agtttagtaa gagagtgatt     3840

ttggctgatg ctaatttgga taaggtgttg agtgcttata ataagcatag agataagcct     3900

attagagagc aggctgagaa tattattcat ttgtttacat tgacaaattt gggagctcct     3960

gctgctttta agtattttga tacaacaatt gatagaaaga gatatacaag tacaaaggag     4020

gtgttggatg ctacattgat tcatcagagt attacaggat tgtatgagac aagaattgat     4080

ttgagtcagt tgggaggaga tggaggagga agtcctaaga agaagagaaa ggtg           4134


<210>  146
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A codons of Table 4 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  146
gacaagaagt actccatcgg cctggacatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggaccacatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga cggcggcggc tcccccaaga agaagcggaa ggtg           4134


<210>  147
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A/U codons of Table 4 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  147
gacaagaagt acagcatcgg cctggacatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggaccacatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga cggcggcggc agccccaaga agaagcggaa ggtg           4134


<210>  148
<211>  4173
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A codons of Table 4, with two 
       C-terminal NLS sequences (no start or stop codons; suitable for 
       inclusion in fusion protein coding sequence)

<400>  148
gacaagaagt actccatcgg cctggacatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggaccacatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga cggctccggc tcccccaaga agaagcggaa ggtggacggc     4140

tcccccaaga agaagcggaa ggtggactcc ggc                                  4173


<210>  149
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A codons of Table 4 (no 
       start or stop codons; suitable for inclusion in fusion protein 
       coding sequence)

<400>  149
gacaagaagt actccatcgg cctggccatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggaccacatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga cggcggcggc tcccccaaga agaagcggaa ggtg           4134


<210>  150
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A codons of Table 4 (no NLS
       and no start or stop codons; suitable for inclusion in fusion 
       protein coding sequence)

<400>  150
gacaagaagt actccatcgg cctggccatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggaccacatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga c                                               4101


<210>  151
<211>  4173
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A codons of Table 4, with 
       two C-terminal NLS sequences (no start or stop codons; suitable 
       for inclusion in fusion protein coding sequence)

<400>  151
gacaagaagt actccatcgg cctggccatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggaccacatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga cggctccggc tcccccaaga agaagcggaa ggtggacggc     4140

tcccccaaga agaagcggaa ggtggactcc ggc                                  4173


<210>  152
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A codons of Table 4 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  152
gacaagaagt actccatcgg cctggccatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggacgccatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga cggcggcggc tcccccaaga agaagcggaa ggtg           4134


<210>  153
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A codons of Table 4 (no NLS and no
       start or stop codons; suitable for inclusion in fusion protein 
       coding sequence)

<400>  153
gacaagaagt actccatcgg cctggccatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggacgccatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga c                                               4101


<210>  154
<211>  4173
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A codons of Table 4, with two 
       C-terminal NLS sequences (no start or stop codons; suitable for 
       inclusion in fusion protein coding sequence)

<400>  154
gacaagaagt actccatcgg cctggccatc ggcaccaact ccgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc ctccaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttctccaa cgagatggcc aaggtggacg actccttctt ccaccggctg      300

gaggagtcct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggact ccaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa ctccgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgcctccg gcgtggacgc caaggccatc ctgtccgccc ggctgtccaa gtcccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctgt ccctgggcct gacccccaac ttcaagtcca acttcgacct ggccgaggac      780

gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga cgccatcctg      900

ctgtccgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgtc cgcctccatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg ctacgccggc     1080

tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg ctccatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caactcccgg     1380

ttcgcctgga tgacccggaa gtccgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgcctccgc ccagtccttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac tccctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgtcc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accggttcaa cgcctccctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgtcccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac catcctggac     2040

ttcctgaagt ccgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgactcc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgtccg gccagggcga ctccctgcac     2160

gagcacatcg ccaacctggc cggctccccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaactcccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggctcccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgtccg actacgacgt ggacgccatc     2520

gtgccccagt ccttcctgaa ggacgactcc atcgacaaca aggtgctgac ccggtccgac     2580

aagaaccggg gcaagtccga caacgtgccc tccgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gtccgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggactcccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagtccaag     2880

ctggtgtccg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agtccgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt ctactccaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgtc catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tctccaagga gtccatcctg cccaagcgga actccgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg actcccccac cgtggcctac     3420

tccgtgctgg tggtggccaa ggtggagaag ggcaagtcca agaagctgaa gtccgtgaag     3480

gagctgctgg gcatcaccat catggagcgg tcctccttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccggaag cggatgctgg cctccgccgg cgagctgcag     3660

aagggcaacg agctggccct gccctccaag tacgtgaact tcctgtacct ggcctcccac     3720

tacgagaagc tgaagggctc ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg tccgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacacctc caccaaggag     4020

gtgctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac ccggatcgac     4080

ctgtcccagc tgggcggcga cggctccggc tcccccaaga agaagcggaa ggtggacggc     4140

tcccccaaga agaagcggaa ggtggactcc ggc                                  4173


<210>  155
<211>  4173
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A/U codons of Table 4, with two 
       C-terminal NLS sequences (no start or stop codons; suitable for 
       inclusion in fusion protein coding sequence)

<400>  155
gacaagaagt acagcatcgg cctggacatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggaccacatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga cggcagcggc agccccaaga agaagcggaa ggtggacggc     4140

agccccaaga agaagcggaa ggtggacagc ggc                                  4173


<210>  156
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 ORF using low A/U codons of Table 4 (no NLS and 
       no start or stop codons; suitable for inclusion in fusion protein
       coding sequence)

<400>  156
gacaagaagt acagcatcgg cctggacatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggaccacatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga c                                               4101


<210>  157
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A/U codons of Table 4 (no 
       start or stop codons; suitable for inclusion in fusion protein 
       coding sequence)

<400>  157
gacaagaagt acagcatcgg cctggccatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggaccacatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga cggcggcggc agccccaaga agaagcggaa ggtg           4134


<210>  158
<211>  4173
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A/U codons of Table 4, with
       two C-terminal NLS sequences (no start or stop codons; suitable 
       for inclusion in fusion protein coding sequence)

<400>  158
gacaagaagt acagcatcgg cctggccatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggaccacatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga cggcagcggc agccccaaga agaagcggaa ggtggacggc     4140

agccccaaga agaagcggaa ggtggacagc ggc                                  4173


<210>  159
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Cas9 nickase ORF using low A/U codons of Table 4 (no 
       NLS and no start or stop codons; suitable for inclusion in fusion
       protein coding sequence)

<400>  159
gacaagaagt acagcatcgg cctggccatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggaccacatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga c                                               4101


<210>  160
<211>  4134
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A/U codons of Table 4 (no start or
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  160
gacaagaagt acagcatcgg cctggccatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggacgccatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga cggcggcggc agccccaaga agaagcggaa ggtg           4134


<210>  161
<211>  4173
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A/U codons of Table 4, with two 
       C-terminal NLS sequences (no start or stop codons; suitable for 
       inclusion in fusion protein coding sequence)

<400>  161
gacaagaagt acagcatcgg cctggccatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggacgccatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga cggcagcggc agccccaaga agaagcggaa ggtggacggc     4140

agccccaaga agaagcggaa ggtggacagc ggc                                  4173


<210>  162
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 ORF using low A/U codons of Table 4 (no NLS and 
       no start or stop codons; suitable for inclusion in fusion protein
       coding sequence)

<400>  162
gacaagaagt acagcatcgg cctggccatc ggcaccaaca gcgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaag ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggcgagac cgccgaggcc      180

acccggctga agcggaccgc ccggcggcgg tacacccggc ggaagaaccg gatctgctac      240

ctgcaggaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccaccggctg      300

gaggagagct tcctggtgga ggaggacaag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gcggaagaag      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct acctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga gaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgagcgccc ggctgagcaa gagccggcgg      660

ctggagaacc tgatcgccca gctgcccggc gagaagaaga acggcctgtt cggcaacctg      720

atcgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggac      780

gccaagctgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgagcga cgccatcctg      900

ctgagcgaca tcctgcgggt gaacaccgag atcaccaagg cccccctgag cgccagcatg      960

atcaagcggt acgacgagca ccaccaggac ctgaccctgc tgaaggccct ggtgcggcag     1020

cagctgcccg agaagtacaa ggagatcttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacatcgacg gcggcgccag ccaggaggag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gctgctggtg aagctgaacc gggaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggcga gctgcacgcc     1260

atcctgcggc ggcaggagga cttctacccc ttcctgaagg acaaccggga gaagatcgag     1320

aagatcctga ccttccggat cccctactac gtgggccccc tggcccgggg caacagccgg     1380

ttcgcctgga tgacccggaa gagcgaggag accatcaccc cctggaactt cgaggaggtg     1440

gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgacaagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc ggaagcccgc cttcctgagc     1620

ggcgagcaga agaaggccat cgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aagcagctga aggaggacta cttcaagaag atcgagtgct tcgacagcgt ggagatcagc     1740

ggcgtggagg accggttcaa cgccagcctg ggcacctacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgggagatg atcgaggagc ggctgaagac ctacgcccac     1920

ctgttcgacg acaaggtgat gaagcagctg aagcggcggc ggtacaccgg ctggggccgg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcaga gcggcaagac catcctggac     2040

ttcctgaaga gcgacggctt cgccaaccgg aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttca aggaggacat ccagaaggcc caggtgagcg gccagggcga cagcctgcac     2160

gagcacatcg ccaacctggc cggcagcccc gccatcaaga agggcatcct gcagaccgtg     2220

aaggtggtgg acgagctggt gaaggtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gagatggccc gggagaacca gaccacccag aagggccaga agaacagccg ggagcggatg     2340

aagcggatcg aggagggcat caaggagctg ggcagccaga tcctgaagga gcaccccgtg     2400

gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa cggccgggac     2460

atgtacgtgg accaggagct ggacatcaac cggctgagcg actacgacgt ggacgccatc     2520

gtgccccaga gcttcctgaa ggacgacagc atcgacaaca aggtgctgac ccggagcgac     2580

aagaaccggg gcaagagcga caacgtgccc agcgaggagg tggtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg atcacccagc ggaagttcga caacctgacc     2700

aaggccgagc ggggcggcct gagcgagctg gacaaggccg gcttcatcaa gcggcagctg     2760

gtggagaccc ggcagatcac caagcacgtg gcccagatcc tggacagccg gatgaacacc     2820

aagtacgacg agaacgacaa gctgatccgg gaggtgaagg tgatcaccct gaagagcaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtggtgggca ccgccctgat caagaagtac     3000

cccaagctgg agagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga gatcggcaag gccaccgcca agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg gaagcggccc     3180

ctgatcgaga ccaacggcga gaccggcgag atcgtgtggg acaagggccg ggacttcgcc     3240

accgtgcgga aggtgctgag catgccccag gtgaacatcg tgaagaagac cgaggtgcag     3300

accggcggct tcagcaagga gagcatcctg cccaagcgga acagcgacaa gctgatcgcc     3360

cggaagaagg actgggaccc caagaagtac ggcggcttcg acagccccac cgtggcctac     3420

agcgtgctgg tggtggccaa ggtggagaag ggcaagagca agaagctgaa gagcgtgaag     3480

gagctgctgg gcatcaccat catggagcgg agcagcttcg agaagaaccc catcgacttc     3540

ctggaggcca agggctacaa ggaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

agcctgttcg agctggagaa cggccggaag cggatgctgg ccagcgccgg cgagctgcag     3660

aagggcaacg agctggccct gcccagcaag tacgtgaact tcctgtacct ggccagccac     3720

tacgagaagc tgaagggcag ccccgaggac aacgagcaga agcagctgtt cgtggagcag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttcagcaa gcgggtgatc     3840

ctggccgacg ccaacctgga caaggtgctg agcgcctaca acaagcaccg ggacaagccc     3900

atccgggagc aggccgagaa catcatccac ctgttcaccc tgaccaacct gggcgccccc     3960

gccgccttca agtacttcga caccaccatc gaccggaagc ggtacaccag caccaaggag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac ccggatcgac     4080

ctgagccagc tgggcggcga c                                               4101


<210>  163
<211>  3306
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Nme Cas9 ORF using low A codons of Table 4 (no start 
       or stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  163
gccgccttca agcccaactc catcaactac atcctgggcc tggacatcgg catcgcctcc       60

gtgggctggg ccatggtgga gatcgacgag gaggagaacc ccatccggct gatcgacctg      120

ggcgtgcggg tgttcgagcg ggccgaggtg cccaagaccg gcgactccct ggccatggcc      180

cggcggctgg cccggtccgt gcggcggctg acccggcggc gggcccaccg gctgctgcgg      240

acccggcggc tgctgaagcg ggagggcgtg ctgcaggccg ccaacttcga cgagaacggc      300

ctgatcaagt ccctgcccaa caccccctgg cagctgcggg ccgccgccct ggaccggaag      360

ctgacccccc tggagtggtc cgccgtgctg ctgcacctga tcaagcaccg gggctacctg      420

tcccagcgga agaacgaggg cgagaccgcc gacaaggagc tgggcgccct gctgaagggc      480

gtggccggca acgcccacgc cctgcagacc ggcgacttcc ggacccccgc cgagctggcc      540

ctgaacaagt tcgagaagga gtccggccac atccggaacc agcggtccga ctactcccac      600

accttctccc ggaaggacct gcaggccgag ctgatcctgc tgttcgagaa gcagaaggag      660

ttcggcaacc cccacgtgtc cggcggcctg aaggagggca tcgagaccct gctgatgacc      720

cagcggcccg ccctgtccgg cgacgccgtg cagaagatgc tgggccactg caccttcgag      780

cccgccgagc ccaaggccgc caagaacacc tacaccgccg agcggttcat ctggctgacc      840

aagctgaaca acctgcggat cctggagcag ggctccgagc ggcccctgac cgacaccgag      900

cgggccaccc tgatggacga gccctaccgg aagtccaagc tgacctacgc ccaggcccgg      960

aagctgctgg gcctggagga caccgccttc ttcaagggcc tgcggtacgg caaggacaac     1020

gccgaggcct ccaccctgat ggagatgaag gcctaccacg ccatctcccg ggccctggag     1080

aaggagggcc tgaaggacaa gaagtccccc ctgaacctgt cccccgagct gcaggacgag     1140

atcggcaccg ccttctccct gttcaagacc gacgaggaca tcaccggccg gctgaaggac     1200

cggatccagc ccgagatcct ggaggccctg ctgaagcaca tctccttcga caagttcgtg     1260

cagatctccc tgaaggccct gcggcggatc gtgcccctga tggagcaggg caagcggtac     1320

gacgaggcct gcgccgagat ctacggcgac cactacggca agaagaacac cgaggagaag     1380

atctacctgc cccccatccc cgccgacgag atccggaacc ccgtggtgct gcgggccctg     1440

tcccaggccc ggaaggtgat caacggcgtg gtgcggcggt acggctcccc cgcccggatc     1500

cacatcgaga ccgcccggga ggtgggcaag tccttcaagg accggaagga gatcgagaag     1560

cggcaggagg agaaccggaa ggaccgggag aaggccgccg ccaagttccg ggagtacttc     1620

cccaacttcg tgggcgagcc caagtccaag gacatcctga agctgcggct gtacgagcag     1680

cagcacggca agtgcctgta ctccggcaag gagatcaacc tgggccggct gaacgagaag     1740

ggctacgtgg agatcgacca cgccctgccc ttctcccgga cctgggacga ctccttcaac     1800

aacaaggtgc tggtgctggg ctccgagaac cagaacaagg gcaaccagac cccctacgag     1860

tacttcaacg gcaaggacaa ctcccgggag tggcaggagt tcaaggcccg ggtggagacc     1920

tcccggttcc cccggtccaa gaagcagcgg atcctgctgc agaagttcga cgaggacggc     1980

ttcaaggagc ggaacctgaa cgacacccgg tacgtgaacc ggttcctgtg ccagttcgtg     2040

gccgaccgga tgcggctgac cggcaagggc aagaagcggg tgttcgcctc caacggccag     2100

atcaccaacc tgctgcgggg cttctggggc ctgcggaagg tgcgggccga gaacgaccgg     2160

caccacgccc tggacgccgt ggtggtggcc tgctccaccg tggccatgca gcagaagatc     2220

acccggttcg tgcggtacaa ggagatgaac gccttcgacg gcaagaccat cgacaaggag     2280

accggcgagg tgctgcacca gaagacccac ttcccccagc cctgggagtt cttcgcccag     2340

gaggtgatga tccgggtgtt cggcaagccc gacggcaagc ccgagttcga ggaggccgac     2400

accctggaga agctgcggac cctgctggcc gagaagctgt cctcccggcc cgaggccgtg     2460

cacgagtacg tgacccccct gttcgtgtcc cgggccccca accggaagat gtccggccag     2520

ggccacatgg agaccgtgaa gtccgccaag cggctggacg agggcgtgtc cgtgctgcgg     2580

gtgcccctga cccagctgaa gctgaaggac ctggagaaga tggtgaaccg ggagcgggag     2640

cccaagctgt acgaggccct gaaggcccgg ctggaggccc acaaggacga ccccgccaag     2700

gccttcgccg agcccttcta caagtacgac aaggccggca accggaccca gcaggtgaag     2760

gccgtgcggg tggagcaggt gcagaagacc ggcgtgtggg tgcggaacca caacggcatc     2820

gccgacaacg ccaccatggt gcgggtggac gtgttcgaga agggcgacaa gtactacctg     2880

gtgcccatct actcctggca ggtggccaag ggcatcctgc ccgaccgggc cgtggtgcag     2940

ggcaaggacg aggaggactg gcagctgatc gacgactcct tcaacttcaa gttctccctg     3000

caccccaacg acctggtgga ggtgatcacc aagaaggccc ggatgttcgg ctacttcgcc     3060

tcctgccacc ggggcaccgg caacatcaac atccggatcc acgacctgga ccacaagatc     3120

ggcaagaacg gcatcctgga gggcatcggc gtgaagaccg ccctgtcctt ccagaagtac     3180

cagatcgacg agctgggcaa ggagatccgg ccctgccggc tgaagaagcg gccccccgtg     3240

cggtccggca agcggaccgc cgacggctcc gagttcgagt cccccaagaa gaagcggaag     3300

gtggag                                                                3306


<210>  164
<211>  3306
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Nme Cas9 ORF using low A/U codons of Table 4 (no start
       or stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  164
gccgccttca agcccaacag catcaactac atcctgggcc tggacatcgg catcgccagc       60

gtgggctggg ccatggtgga gatcgacgag gaggagaacc ccatccggct gatcgacctg      120

ggcgtgcggg tgttcgagcg ggccgaggtg cccaagaccg gcgacagcct ggccatggcc      180

cggcggctgg cccggagcgt gcggcggctg acccggcggc gggcccaccg gctgctgcgg      240

acccggcggc tgctgaagcg ggagggcgtg ctgcaggccg ccaacttcga cgagaacggc      300

ctgatcaaga gcctgcccaa caccccctgg cagctgcggg ccgccgccct ggaccggaag      360

ctgacccccc tggagtggag cgccgtgctg ctgcacctga tcaagcaccg gggctacctg      420

agccagcgga agaacgaggg cgagaccgcc gacaaggagc tgggcgccct gctgaagggc      480

gtggccggca acgcccacgc cctgcagacc ggcgacttcc ggacccccgc cgagctggcc      540

ctgaacaagt tcgagaagga gagcggccac atccggaacc agcggagcga ctacagccac      600

accttcagcc ggaaggacct gcaggccgag ctgatcctgc tgttcgagaa gcagaaggag      660

ttcggcaacc cccacgtgag cggcggcctg aaggagggca tcgagaccct gctgatgacc      720

cagcggcccg ccctgagcgg cgacgccgtg cagaagatgc tgggccactg caccttcgag      780

cccgccgagc ccaaggccgc caagaacacc tacaccgccg agcggttcat ctggctgacc      840

aagctgaaca acctgcggat cctggagcag ggcagcgagc ggcccctgac cgacaccgag      900

cgggccaccc tgatggacga gccctaccgg aagagcaagc tgacctacgc ccaggcccgg      960

aagctgctgg gcctggagga caccgccttc ttcaagggcc tgcggtacgg caaggacaac     1020

gccgaggcca gcaccctgat ggagatgaag gcctaccacg ccatcagccg ggccctggag     1080

aaggagggcc tgaaggacaa gaagagcccc ctgaacctga gccccgagct gcaggacgag     1140

atcggcaccg ccttcagcct gttcaagacc gacgaggaca tcaccggccg gctgaaggac     1200

cggatccagc ccgagatcct ggaggccctg ctgaagcaca tcagcttcga caagttcgtg     1260

cagatcagcc tgaaggccct gcggcggatc gtgcccctga tggagcaggg caagcggtac     1320

gacgaggcct gcgccgagat ctacggcgac cactacggca agaagaacac cgaggagaag     1380

atctacctgc cccccatccc cgccgacgag atccggaacc ccgtggtgct gcgggccctg     1440

agccaggccc ggaaggtgat caacggcgtg gtgcggcggt acggcagccc cgcccggatc     1500

cacatcgaga ccgcccggga ggtgggcaag agcttcaagg accggaagga gatcgagaag     1560

cggcaggagg agaaccggaa ggaccgggag aaggccgccg ccaagttccg ggagtacttc     1620

cccaacttcg tgggcgagcc caagagcaag gacatcctga agctgcggct gtacgagcag     1680

cagcacggca agtgcctgta cagcggcaag gagatcaacc tgggccggct gaacgagaag     1740

ggctacgtgg agatcgacca cgccctgccc ttcagccgga cctgggacga cagcttcaac     1800

aacaaggtgc tggtgctggg cagcgagaac cagaacaagg gcaaccagac cccctacgag     1860

tacttcaacg gcaaggacaa cagccgggag tggcaggagt tcaaggcccg ggtggagacc     1920

agccggttcc cccggagcaa gaagcagcgg atcctgctgc agaagttcga cgaggacggc     1980

ttcaaggagc ggaacctgaa cgacacccgg tacgtgaacc ggttcctgtg ccagttcgtg     2040

gccgaccgga tgcggctgac cggcaagggc aagaagcggg tgttcgccag caacggccag     2100

atcaccaacc tgctgcgggg cttctggggc ctgcggaagg tgcgggccga gaacgaccgg     2160

caccacgccc tggacgccgt ggtggtggcc tgcagcaccg tggccatgca gcagaagatc     2220

acccggttcg tgcggtacaa ggagatgaac gccttcgacg gcaagaccat cgacaaggag     2280

accggcgagg tgctgcacca gaagacccac ttcccccagc cctgggagtt cttcgcccag     2340

gaggtgatga tccgggtgtt cggcaagccc gacggcaagc ccgagttcga ggaggccgac     2400

accctggaga agctgcggac cctgctggcc gagaagctga gcagccggcc cgaggccgtg     2460

cacgagtacg tgacccccct gttcgtgagc cgggccccca accggaagat gagcggccag     2520

ggccacatgg agaccgtgaa gagcgccaag cggctggacg agggcgtgag cgtgctgcgg     2580

gtgcccctga cccagctgaa gctgaaggac ctggagaaga tggtgaaccg ggagcgggag     2640

cccaagctgt acgaggccct gaaggcccgg ctggaggccc acaaggacga ccccgccaag     2700

gccttcgccg agcccttcta caagtacgac aaggccggca accggaccca gcaggtgaag     2760

gccgtgcggg tggagcaggt gcagaagacc ggcgtgtggg tgcggaacca caacggcatc     2820

gccgacaacg ccaccatggt gcgggtggac gtgttcgaga agggcgacaa gtactacctg     2880

gtgcccatct acagctggca ggtggccaag ggcatcctgc ccgaccgggc cgtggtgcag     2940

ggcaaggacg aggaggactg gcagctgatc gacgacagct tcaacttcaa gttcagcctg     3000

caccccaacg acctggtgga ggtgatcacc aagaaggccc ggatgttcgg ctacttcgcc     3060

agctgccacc ggggcaccgg caacatcaac atccggatcc acgacctgga ccacaagatc     3120

ggcaagaacg gcatcctgga gggcatcggc gtgaagaccg ccctgagctt ccagaagtac     3180

cagatcgacg agctgggcaa ggagatccgg ccctgccggc tgaagaagcg gccccccgtg     3240

cggagcggca agcggaccgc cgacggcagc gagttcgaga gccccaagaa gaagcggaag     3300

gtggag                                                                3306


<210>  165
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS1 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  165
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcctggcag caaagagaag cagaacaaca     4140


<210>  166
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS2 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  166
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agccaggcag caaagagaag cagaacaaca     4140


<210>  167
<211>  4143
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS3 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  167
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcccggcac cggcaaagag agaaagaaca     4140

aca                                                                   4143


<210>  168
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS4 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  168
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agccaggcag caaagagacc gagaacaaca     4140


<210>  169
<211>  4140
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS5 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  169
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcagagcag caaagagacc gagaacaaca     4140


<210>  170
<211>  4146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS6 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  170
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcgcagcag caaagagaag ctggagcatg     4140

gcagca                                                                4146


<210>  171
<211>  4146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS7 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  171
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcgcagcag caaagagagt ctggagcatg     4140

gcattc                                                                4146


<210>  172
<211>  4146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS8 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  172
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcgcagcag caaagagaag ctggagcatg     4140

gcattc                                                                4146


<210>  173
<211>  4143
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS9 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  173
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcgcagcag caaagagaaa gtacttcgca     4140

gca                                                                   4143


<210>  174
<211>  4143
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS10 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  174
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcagagcag caaagagaaa ggcattcgca     4140

gca                                                                   4143


<210>  175
<211>  4143
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Open reading frame for Cas9 with NLS11 (no start or 
       stop codons; suitable for inclusion in fusion protein coding 
       sequence)

<400>  175
gacaagaagt acagcatcgg actggacatc ggaacaaaca gcgtcggatg ggcagtcatc       60

acagacgaat acaaggtccc gagcaagaag ttcaaggtcc tgggaaacac agacagacac      120

agcatcaaga agaacctgat cggagcactg ctgttcgaca gcggagaaac agcagaagca      180

acaagactga agagaacagc aagaagaaga tacacaagaa gaaagaacag aatctgctac      240

ctgcaggaaa tcttcagcaa cgaaatggca aaggtcgacg acagcttctt ccacagactg      300

gaagaaagct tcctggtcga agaagacaag aagcacgaaa gacacccgat cttcggaaac      360

atcgtcgacg aagtcgcata ccacgaaaag tacccgacaa tctaccacct gagaaagaag      420

ctggtcgaca gcacagacaa ggcagacctg agactgatct acctggcact ggcacacatg      480

atcaagttca gaggacactt cctgatcgaa ggagacctga acccggacaa cagcgacgtc      540

gacaagctgt tcatccagct ggtccagaca tacaaccagc tgttcgaaga aaacccgatc      600

aacgcaagcg gagtcgacgc aaaggcaatc ctgagcgcaa gactgagcaa gagcagaaga      660

ctggaaaacc tgatcgcaca gctgccggga gaaaagaaga acggactgtt cggaaacctg      720

atcgcactga gcctgggact gacaccgaac ttcaagagca acttcgacct ggcagaagac      780

gcaaagctgc agctgagcaa ggacacatac gacgacgacc tggacaacct gctggcacag      840

atcggagacc agtacgcaga cctgttcctg gcagcaaaga acctgagcga cgcaatcctg      900

ctgagcgaca tcctgagagt caacacagaa atcacaaagg caccgctgag cgcaagcatg      960

atcaagagat acgacgaaca ccaccaggac ctgacactgc tgaaggcact ggtcagacag     1020

cagctgccgg aaaagtacaa ggaaatcttc ttcgaccaga gcaagaacgg atacgcagga     1080

tacatcgacg gaggagcaag ccaggaagaa ttctacaagt tcatcaagcc gatcctggaa     1140

aagatggacg gaacagaaga actgctggtc aagctgaaca gagaagacct gctgagaaag     1200

cagagaacat tcgacaacgg aagcatcccg caccagatcc acctgggaga actgcacgca     1260

atcctgagaa gacaggaaga cttctacccg ttcctgaagg acaacagaga aaagatcgaa     1320

aagatcctga cattcagaat cccgtactac gtcggaccgc tggcaagagg aaacagcaga     1380

ttcgcatgga tgacaagaaa gagcgaagaa acaatcacac cgtggaactt cgaagaagtc     1440

gtcgacaagg gagcaagcgc acagagcttc atcgaaagaa tgacaaactt cgacaagaac     1500

ctgccgaacg aaaaggtcct gccgaagcac agcctgctgt acgaatactt cacagtctac     1560

aacgaactga caaaggtcaa gtacgtcaca gaaggaatga gaaagccggc attcctgagc     1620

ggagaacaga agaaggcaat cgtcgacctg ctgttcaaga caaacagaaa ggtcacagtc     1680

aagcagctga aggaagacta cttcaagaag atcgaatgct tcgacagcgt cgaaatcagc     1740

ggagtcgaag acagattcaa cgcaagcctg ggaacatacc acgacctgct gaagatcatc     1800

aaggacaagg acttcctgga caacgaagaa aacgaagaca tcctggaaga catcgtcctg     1860

acactgacac tgttcgaaga cagagaaatg atcgaagaaa gactgaagac atacgcacac     1920

ctgttcgacg acaaggtcat gaagcagctg aagagaagaa gatacacagg atggggaaga     1980

ctgagcagaa agctgatcaa cggaatcaga gacaagcaga gcggaaagac aatcctggac     2040

ttcctgaaga gcgacggatt cgcaaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgacattca aggaagacat ccagaaggca caggtcagcg gacagggaga cagcctgcac     2160

gaacacatcg caaacctggc aggaagcccg gcaatcaaga agggaatcct gcagacagtc     2220

aaggtcgtcg acgaactggt caaggtcatg ggaagacaca agccggaaaa catcgtcatc     2280

gaaatggcaa gagaaaacca gacaacacag aagggacaga agaacagcag agaaagaatg     2340

aagagaatcg aagaaggaat caaggaactg ggaagccaga tcctgaagga acacccggtc     2400

gaaaacacac agctgcagaa cgaaaagctg tacctgtact acctgcagaa cggaagagac     2460

atgtacgtcg accaggaact ggacatcaac agactgagcg actacgacgt cgaccacatc     2520

gtcccgcaga gcttcctgaa ggacgacagc atcgacaaca aggtcctgac aagaagcgac     2580

aagaacagag gaaagagcga caacgtcccg agcgaagaag tcgtcaagaa gatgaagaac     2640

tactggagac agctgctgaa cgcaaagctg atcacacaga gaaagttcga caacctgaca     2700

aaggcagaga gaggaggact gagcgaactg gacaaggcag gattcatcaa gagacagctg     2760

gtcgaaacaa gacagatcac aaagcacgtc gcacagatcc tggacagcag aatgaacaca     2820

aagtacgacg aaaacgacaa gctgatcaga gaagtcaagg tcatcacact gaagagcaag     2880

ctggtcagcg acttcagaaa ggacttccag ttctacaagg tcagagaaat caacaactac     2940

caccacgcac acgacgcata cctgaacgca gtcgtcggaa cagcactgat caagaagtac     3000

ccgaagctgg aaagcgaatt cgtctacgga gactacaagg tctacgacgt cagaaagatg     3060

atcgcaaaga gcgaacagga aatcggaaag gcaacagcaa agtacttctt ctacagcaac     3120

atcatgaact tcttcaagac agaaatcaca ctggcaaacg gagaaatcag aaagagaccg     3180

ctgatcgaaa caaacggaga aacaggagaa atcgtctggg acaagggaag agacttcgca     3240

acagtcagaa aggtcctgag catgccgcag gtcaacatcg tcaagaagac agaagtccag     3300

acaggaggat tcagcaagga aagcatcctg ccgaagagaa acagcgacaa gctgatcgca     3360

agaaagaagg actgggaccc gaagaagtac ggaggattcg acagcccgac agtcgcatac     3420

agcgtcctgg tcgtcgcaaa ggtcgaaaag ggaaagagca agaagctgaa gagcgtcaag     3480

gaactgctgg gaatcacaat catggaaaga agcagcttcg aaaagaaccc gatcgacttc     3540

ctggaagcaa agggatacaa ggaagtcaag aaggacctga tcatcaagct gccgaagtac     3600

agcctgttcg aactggaaaa cggaagaaag agaatgctgg caagcgcagg agaactgcag     3660

aagggaaacg aactggcact gccgagcaag tacgtcaact tcctgtacct ggcaagccac     3720

tacgaaaagc tgaagggaag cccggaagac aacgaacaga agcagctgtt cgtcgaacag     3780

cacaagcact acctggacga aatcatcgaa cagatcagcg aattcagcaa gagagtcatc     3840

ctggcagacg caaacctgga caaggtcctg agcgcataca acaagcacag agacaagccg     3900

atcagagaac aggcagaaaa catcatccac ctgttcacac tgacaaacct gggagcaccg     3960

gcagcattca agtacttcga cacaacaatc gacagaaaga gatacacaag cacaaaggaa     4020

gtcctggacg caacactgat ccaccagagc atcacaggac tgtacgaaac aagaatcgac     4080

ctgagccagc tgggaggaga cggaggagga agcagagcag caaagagaaa gtacttcgca     4140

gtc                                                                   4143


<210>  176
<211>  4423
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with XBG UTRs and Cas9 ORF with low U 
       1 codons of Table 4

<400>  176
gggaagcuca gaauaaacgc ucaacuuugg ccggaucugc caccauggac aagaaguaca       60

gcaucggccu ggacaucggc accaacagcg ugggcugggc cgugaucacc gacgaguaca      120

aggugcccag caagaaguuc aaggugcugg gcaacaccga cagacacagc aucaagaaga      180

accugaucgg cgcccugcug uucgacagcg gcgagaccgc cgaggccacc agacugaaga      240

gaaccgccag aagaagauac accagaagaa agaacagaau cugcuaccug caggagaucu      300

ucagcaacga gauggccaag guggacgaca gcuucuucca cagacuggag gagagcuucc      360

ugguggagga ggacaagaag cacgagagac accccaucuu cggcaacauc guggacgagg      420

uggccuacca cgagaaguac cccaccaucu accaccugag aaagaagcug guggacagca      480

ccgacaaggc cgaccugaga cugaucuacc uggcccuggc ccacaugauc aaguucagag      540

gccacuuccu gaucgagggc gaccugaacc ccgacaacag cgacguggac aagcuguuca      600

uccagcuggu gcagaccuac aaccagcugu ucgaggagaa ccccaucaac gccagcggcg      660

uggacgccaa ggccauccug agcgccagac ugagcaagag cagaagacug gagaaccuga      720

ucgcccagcu gcccggcgag aagaagaacg gccuguucgg caaccugauc gcccugagcc      780

ugggccugac ccccaacuuc aagagcaacu ucgaccuggc cgaggacgcc aagcugcagc      840

ugagcaagga caccuacgac gacgaccugg acaaccugcu ggcccagauc ggcgaccagu      900

acgccgaccu guuccuggcc gccaagaacc ugagcgacgc cauccugcug agcgacaucc      960

ugagagugaa caccgagauc accaaggccc cccugagcgc cagcaugauc aagagauacg     1020

acgagcacca ccaggaccug acccugcuga aggcccuggu gagacagcag cugcccgaga     1080

aguacaagga gaucuucuuc gaccagagca agaacggcua cgccggcuac aucgacggcg     1140

gcgccagcca ggaggaguuc uacaaguuca ucaagcccau ccuggagaag auggacggca     1200

ccgaggagcu gcuggugaag cugaacagag aggaccugcu gagaaagcag agaaccuucg     1260

acaacggcag caucccccac cagauccacc ugggcgagcu gcacgccauc cugagaagac     1320

aggaggacuu cuaccccuuc cugaaggaca acagagagaa gaucgagaag auccugaccu     1380

ucagaauccc cuacuacgug ggcccccugg ccagaggcaa cagcagauuc gccuggauga     1440

ccagaaagag cgaggagacc aucacccccu ggaacuucga ggagguggug gacaagggcg     1500

ccagcgccca gagcuucauc gagagaauga ccaacuucga caagaaccug cccaacgaga     1560

aggugcugcc caagcacagc cugcuguacg aguacuucac cguguacaac gagcugacca     1620

aggugaagua cgugaccgag ggcaugagaa agcccgccuu ccugagcggc gagcagaaga     1680

aggccaucgu ggaccugcug uucaagacca acagaaaggu gaccgugaag cagcugaagg     1740

aggacuacuu caagaagauc gagugcuucg acagcgugga gaucagcggc guggaggaca     1800

gauucaacgc cagccugggc accuaccacg accugcugaa gaucaucaag gacaaggacu     1860

uccuggacaa cgaggagaac gaggacaucc uggaggacau cgugcugacc cugacccugu     1920

ucgaggacag agagaugauc gaggagagac ugaagaccua cgcccaccug uucgacgaca     1980

aggugaugaa gcagcugaag agaagaagau acaccggcug gggcagacug agcagaaagc     2040

ugaucaacgg caucagagac aagcagagcg gcaagaccau ccuggacuuc cugaagagcg     2100

acggcuucgc caacagaaac uucaugcagc ugauccacga cgacagccug accuucaagg     2160

aggacaucca gaaggcccag gugagcggcc agggcgacag ccugcacgag cacaucgcca     2220

accuggccgg cagccccgcc aucaagaagg gcauccugca gaccgugaag gugguggacg     2280

agcuggugaa ggugaugggc agacacaagc ccgagaacau cgugaucgag auggccagag     2340

agaaccagac cacccagaag ggccagaaga acagcagaga gagaaugaag agaaucgagg     2400

agggcaucaa ggagcugggc agccagaucc ugaaggagca ccccguggag aacacccagc     2460

ugcagaacga gaagcuguac cuguacuacc ugcagaacgg cagagacaug uacguggacc     2520

aggagcugga caucaacaga cugagcgacu acgacgugga ccacaucgug ccccagagcu     2580

uccugaagga cgacagcauc gacaacaagg ugcugaccag aagcgacaag aacagaggca     2640

agagcgacaa cgugcccagc gaggaggugg ugaagaagau gaagaacuac uggagacagc     2700

ugcugaacgc caagcugauc acccagagaa aguucgacaa ccugaccaag gccgagagag     2760

gcggccugag cgagcuggac aaggccggcu ucaucaagag acagcuggug gagaccagac     2820

agaucaccaa gcacguggcc cagauccugg acagcagaau gaacaccaag uacgacgaga     2880

acgacaagcu gaucagagag gugaagguga ucacccugaa gagcaagcug gugagcgacu     2940

ucagaaagga cuuccaguuc uacaagguga gagagaucaa caacuaccac cacgcccacg     3000

acgccuaccu gaacgccgug gugggcaccg cccugaucaa gaaguacccc aagcuggaga     3060

gcgaguucgu guacggcgac uacaaggugu acgacgugag aaagaugauc gccaagagcg     3120

agcaggagau cggcaaggcc accgccaagu acuucuucua cagcaacauc augaacuucu     3180

ucaagaccga gaucacccug gccaacggcg agaucagaaa gagaccccug aucgagacca     3240

acggcgagac cggcgagauc gugugggaca agggcagaga cuucgccacc gugagaaagg     3300

ugcugagcau gccccaggug aacaucguga agaagaccga ggugcagacc ggcggcuuca     3360

gcaaggagag cauccugccc aagagaaaca gcgacaagcu gaucgccaga aagaaggacu     3420

gggaccccaa gaaguacggc ggcuucgaca gccccaccgu ggccuacagc gugcuggugg     3480

uggccaaggu ggagaagggc aagagcaaga agcugaagag cgugaaggag cugcugggca     3540

ucaccaucau ggagagaagc agcuucgaga agaaccccau cgacuuccug gaggccaagg     3600

gcuacaagga ggugaagaag gaccugauca ucaagcugcc caaguacagc cuguucgagc     3660

uggagaacgg cagaaagaga augcuggcca gcgccggcga gcugcagaag ggcaacgagc     3720

uggcccugcc cagcaaguac gugaacuucc uguaccuggc cagccacuac gagaagcuga     3780

agggcagccc cgaggacaac gagcagaagc agcuguucgu ggagcagcac aagcacuacc     3840

uggacgagau caucgagcag aucagcgagu ucagcaagag agugauccug gccgacgcca     3900

accuggacaa ggugcugagc gccuacaaca agcacagaga caagcccauc agagagcagg     3960

ccgagaacau cauccaccug uucacccuga ccaaccuggg cgcccccgcc gccuucaagu     4020

acuucgacac caccaucgac agaaagagau acaccagcac caaggaggug cuggacgcca     4080

cccugaucca ccagagcauc accggccugu acgagaccag aaucgaccug agccagcugg     4140

gcggcgacgg cggcggcagc cccaagaaga agagaaaggu gugacuagca ccagccucaa     4200

gaacacccga auggagucuc uaagcuacau aauaccaacu uacacuuuac aaaauguugu     4260

cccccaaaau guagccauuc guaucugcuc cuaauaaaaa gaaaguuucu ucacauucuc     4320

ucgagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4380

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaauc uag                       4423


<210>  177
<211>  4423
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with XBG UTRs and Cas9 ORF with low A 
       codons of Table 4

<400>  177
gggaagcuca gaauaaacgc ucaacuuugg ccggaucugc caccauggac aagaaguacu       60

ccaucggccu ggacaucggc accaacuccg ugggcugggc cgugaucacc gacgaguaca      120

aggugcccuc caagaaguuc aaggugcugg gcaacaccga ccggcacucc aucaagaaga      180

accugaucgg cgcccugcug uucgacuccg gcgagaccgc cgaggccacc cggcugaagc      240

ggaccgcccg gcggcgguac acccggcgga agaaccggau cugcuaccug caggagaucu      300

ucuccaacga gauggccaag guggacgacu ccuucuucca ccggcuggag gaguccuucc      360

ugguggagga ggacaagaag cacgagcggc accccaucuu cggcaacauc guggacgagg      420

uggccuacca cgagaaguac cccaccaucu accaccugcg gaagaagcug guggacucca      480

ccgacaaggc cgaccugcgg cugaucuacc uggcccuggc ccacaugauc aaguuccggg      540

gccacuuccu gaucgagggc gaccugaacc ccgacaacuc cgacguggac aagcuguuca      600

uccagcuggu gcagaccuac aaccagcugu ucgaggagaa ccccaucaac gccuccggcg      660

uggacgccaa ggccauccug uccgcccggc uguccaaguc ccggcggcug gagaaccuga      720

ucgcccagcu gcccggcgag aagaagaacg gccuguucgg caaccugauc gcccuguccc      780

ugggccugac ccccaacuuc aaguccaacu ucgaccuggc cgaggacgcc aagcugcagc      840

uguccaagga caccuacgac gacgaccugg acaaccugcu ggcccagauc ggcgaccagu      900

acgccgaccu guuccuggcc gccaagaacc uguccgacgc cauccugcug uccgacaucc      960

ugcgggugaa caccgagauc accaaggccc cccuguccgc cuccaugauc aagcgguacg     1020

acgagcacca ccaggaccug acccugcuga aggcccuggu gcggcagcag cugcccgaga     1080

aguacaagga gaucuucuuc gaccagucca agaacggcua cgccggcuac aucgacggcg     1140

gcgccuccca ggaggaguuc uacaaguuca ucaagcccau ccuggagaag auggacggca     1200

ccgaggagcu gcuggugaag cugaaccggg aggaccugcu gcggaagcag cggaccuucg     1260

acaacggcuc caucccccac cagauccacc ugggcgagcu gcacgccauc cugcggcggc     1320

aggaggacuu cuaccccuuc cugaaggaca accgggagaa gaucgagaag auccugaccu     1380

uccggauccc cuacuacgug ggcccccugg cccggggcaa cucccgguuc gccuggauga     1440

cccggaaguc cgaggagacc aucacccccu ggaacuucga ggagguggug gacaagggcg     1500

ccuccgccca guccuucauc gagcggauga ccaacuucga caagaaccug cccaacgaga     1560

aggugcugcc caagcacucc cugcuguacg aguacuucac cguguacaac gagcugacca     1620

aggugaagua cgugaccgag ggcaugcgga agcccgccuu ccuguccggc gagcagaaga     1680

aggccaucgu ggaccugcug uucaagacca accggaaggu gaccgugaag cagcugaagg     1740

aggacuacuu caagaagauc gagugcuucg acuccgugga gaucuccggc guggaggacc     1800

gguucaacgc cucccugggc accuaccacg accugcugaa gaucaucaag gacaaggacu     1860

uccuggacaa cgaggagaac gaggacaucc uggaggacau cgugcugacc cugacccugu     1920

ucgaggaccg ggagaugauc gaggagcggc ugaagaccua cgcccaccug uucgacgaca     1980

aggugaugaa gcagcugaag cggcggcggu acaccggcug gggccggcug ucccggaagc     2040

ugaucaacgg cauccgggac aagcaguccg gcaagaccau ccuggacuuc cugaaguccg     2100

acggcuucgc caaccggaac uucaugcagc ugauccacga cgacucccug accuucaagg     2160

aggacaucca gaaggcccag guguccggcc agggcgacuc ccugcacgag cacaucgcca     2220

accuggccgg cucccccgcc aucaagaagg gcauccugca gaccgugaag gugguggacg     2280

agcuggugaa ggugaugggc cggcacaagc ccgagaacau cgugaucgag auggcccggg     2340

agaaccagac cacccagaag ggccagaaga acucccggga gcggaugaag cggaucgagg     2400

agggcaucaa ggagcugggc ucccagaucc ugaaggagca ccccguggag aacacccagc     2460

ugcagaacga gaagcuguac cuguacuacc ugcagaacgg ccgggacaug uacguggacc     2520

aggagcugga caucaaccgg cuguccgacu acgacgugga ccacaucgug ccccaguccu     2580

uccugaagga cgacuccauc gacaacaagg ugcugacccg guccgacaag aaccggggca     2640

aguccgacaa cgugcccucc gaggaggugg ugaagaagau gaagaacuac uggcggcagc     2700

ugcugaacgc caagcugauc acccagcgga aguucgacaa ccugaccaag gccgagcggg     2760

gcggccuguc cgagcuggac aaggccggcu ucaucaagcg gcagcuggug gagacccggc     2820

agaucaccaa gcacguggcc cagauccugg acucccggau gaacaccaag uacgacgaga     2880

acgacaagcu gauccgggag gugaagguga ucacccugaa guccaagcug guguccgacu     2940

uccggaagga cuuccaguuc uacaaggugc gggagaucaa caacuaccac cacgcccacg     3000

acgccuaccu gaacgccgug gugggcaccg cccugaucaa gaaguacccc aagcuggagu     3060

ccgaguucgu guacggcgac uacaaggugu acgacgugcg gaagaugauc gccaaguccg     3120

agcaggagau cggcaaggcc accgccaagu acuucuucua cuccaacauc augaacuucu     3180

ucaagaccga gaucacccug gccaacggcg agauccggaa gcggccccug aucgagacca     3240

acggcgagac cggcgagauc gugugggaca agggccggga cuucgccacc gugcggaagg     3300

ugcuguccau gccccaggug aacaucguga agaagaccga ggugcagacc ggcggcuucu     3360

ccaaggaguc cauccugccc aagcggaacu ccgacaagcu gaucgcccgg aagaaggacu     3420

gggaccccaa gaaguacggc ggcuucgacu cccccaccgu ggccuacucc gugcuggugg     3480

uggccaaggu ggagaagggc aaguccaaga agcugaaguc cgugaaggag cugcugggca     3540

ucaccaucau ggagcggucc uccuucgaga agaaccccau cgacuuccug gaggccaagg     3600

gcuacaagga ggugaagaag gaccugauca ucaagcugcc caaguacucc cuguucgagc     3660

uggagaacgg ccggaagcgg augcuggccu ccgccggcga gcugcagaag ggcaacgagc     3720

uggcccugcc cuccaaguac gugaacuucc uguaccuggc cucccacuac gagaagcuga     3780

agggcucccc cgaggacaac gagcagaagc agcuguucgu ggagcagcac aagcacuacc     3840

uggacgagau caucgagcag aucuccgagu ucuccaagcg ggugauccug gccgacgcca     3900

accuggacaa ggugcugucc gccuacaaca agcaccggga caagcccauc cgggagcagg     3960

ccgagaacau cauccaccug uucacccuga ccaaccuggg cgcccccgcc gccuucaagu     4020

acuucgacac caccaucgac cggaagcggu acaccuccac caaggaggug cuggacgcca     4080

cccugaucca ccaguccauc accggccugu acgagacccg gaucgaccug ucccagcugg     4140

gcggcgacgg cggcggcucc cccaagaaga agcggaaggu gugacuagca ccagccucaa     4200

gaacacccga auggagucuc uaagcuacau aauaccaacu uacacuuuac aaaauguugu     4260

cccccaaaau guagccauuc guaucugcuc cuaauaaaaa gaaaguuucu ucacauucuc     4320

ucgagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4380

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaauc uag                       4423


<210>  178
<211>  4421
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with XBG UTRs and Cas9 ORF with low 
       U/A codons of Table 4

<400>  178
gggaagcuca gaauaaacgc ucaacuuugg ccggaucugc caccauggac aagaaguaca       60

gcaucggccu ggacaucggc accaacagcg ugggcugggc cgugaucacc gacgaguaca      120

aggugcccag caagaaguuc aaggugcugg gcaacaccga ccggcacagc aucaagaaga      180

accugaucgg cgcccugcug uucgacagcg gcgagaccgc cgaggccacc cggcugaagc      240

ggaccgcccg gcggcgguac acccggcgga agaaccggau cugcuaccug caggagaucu      300

ucagcaacga gauggccaag guggacgaca gcuucuucca ccggcuggag gagagcuucc      360

ugguggagga ggacaagaag cacgagcggc accccaucuu cggcaacauc guggacgagg      420

uggccuacca cgagaaguac cccaccaucu accaccugcg gaagaagcug guggacagca      480

ccgacaaggc cgaccugcgg cugaucuacc uggcccuggc ccacaugauc aaguuccggg      540

gccacuuccu gaucgagggc gaccugaacc ccgacaacag cgacguggac aagcuguuca      600

uccagcuggu gcagaccuac aaccagcugu ucgaggagaa ccccaucaac gccagcggcg      660

uggacgccaa ggccauccug agcgcccggc ugagcaagag ccggcggcug gagaaccuga      720

ucgcccagcu gcccggcgag aagaagaacg gccuguucgg caaccugauc gcccugagcc      780

ugggccugac ccccaacuuc aagagcaacu ucgaccuggc cgaggacgcc aagcugcagc      840

ugagcaagga caccuacgac gacgaccugg acaaccugcu ggcccagauc ggcgaccagu      900

acgccgaccu guuccuggcc gccaagaacc ugagcgacgc cauccugcug agcgacaucc      960

ugcgggugaa caccgagauc accaaggccc cccugagcgc cagcaugauc aagcgguacg     1020

acgagcacca ccaggaccug acccugcuga aggcccuggu gcggcagcag cugcccgaga     1080

aguacaagga gaucuucuuc gaccagagca agaacggcua cgccggcuac aucgacggcg     1140

gcgccagcca ggaggaguuc uacaaguuca ucaagcccau ccuggagaag auggacggca     1200

ccgaggagcu gcuggugaag cugaaccggg aggaccugcu gcggaagcag cggaccuucg     1260

acaacggcag caucccccac cagauccacc ugggcgagcu gcacgccauc cugcggcggc     1320

aggaggacuu cuaccccuuc cugaaggaca accgggagaa gaucgagaag auccugaccu     1380

uccggauccc cuacuacgug ggcccccugg cccggggcaa cagccgguuc gccuggauga     1440

cccggaagag cgaggagacc aucacccccu ggaacuucga ggagguggug gacaagggcg     1500

ccagcgccca gagcuucauc gagcggauga ccaacuucga caagaaccug cccaacgaga     1560

aggugcugcc caagcacagc cugcuguacg aguacuucac cguguacaac gagcugacca     1620

aggugaagua cgugaccgag ggcaugcgga agcccgccuu ccugagcggc gagcagaaga     1680

aggccaucgu ggaccugcug uucaagacca accggaaggu gaccgugaag cagcugaagg     1740

aggacuacuu caagaagauc gagugcuucg acagcgugga gaucagcggc guggaggacc     1800

gguucaacgc cagccugggc accuaccacg accugcugaa gaucaucaag gacaaggacu     1860

uccuggacaa cgaggagaac gaggacaucc uggaggacau cgugcugacc cugacccugu     1920

ucgaggaccg ggagaugauc gaggagcggc ugaagaccua cgcccaccug uucgacgaca     1980

aggugaugaa gcagcugaag cggcggcggu acaccggcug gggccggcug agccggaagc     2040

ugaucaacgg cauccgggac aagcagagcg gcaagaccau ccuggacuuc cugaagagcg     2100

acggcuucgc caaccggaac uucaugcagc ugauccacga cgacagccug accuucaagg     2160

aggacaucca gaaggcccag gugagcggcc agggcgacag ccugcacgag cacaucgcca     2220

accuggccgg cagccccgcc aucaagaagg gcauccugca gaccgugaag gugguggacg     2280

agcuggugaa ggugaugggc cggcacaagc ccgagaacau cgugaucgag auggcccggg     2340

agaaccagac cacccagaag ggccagaaga acagccggga gcggaugaag cggaucgagg     2400

agggcaucaa ggagcugggc agccagaucc ugaaggagca ccccguggag aacacccagc     2460

ugcagaacga gaagcuguac cuguacuacc ugcagaacgg ccgggacaug uacguggacc     2520

aggagcugga caucaaccgg cugagcgacu acgacgugga ccacaucgug ccccagagcu     2580

uccugaagga cgacagcauc gacaacaagg ugcugacccg gagcgacaag aaccggggca     2640

agagcgacaa cgugcccagc gaggaggugg ugaagaagau gaagaacuac uggcggcagc     2700

ugcugaacgc caagcugauc acccagcgga aguucgacaa ccugaccaag gccgagcggg     2760

gcggccugag cgagcuggac aaggccggcu ucaucaagcg gcagcuggug gagacccggc     2820

agaucaccaa gcacguggcc cagauccugg acagccggau gaacaccaag uacgacgaga     2880

acgacaagcu gauccgggag gugaagguga ucacccugaa gagcaagcug gugagcgacu     2940

uccggaagga cuuccaguuc uacaaggugc gggagaucaa caacuaccac cacgcccacg     3000

acgccuaccu gaacgccgug gugggcaccg cccugaucaa gaaguacccc aagcuggaga     3060

gcgaguucgu guacggcgac uacaaggugu acgacgugcg gaagaugauc gccaagagcg     3120

agcaggagau cggcaaggcc accgccaagu acuucuucua cagcaacauc augaacuucu     3180

ucaagaccga gaucacccug gccaacggcg agauccggaa gcggccccug aucgagacca     3240

acggcgagac cggcgagauc gugugggaca agggccggga cuucgccacc gugcggaagg     3300

ugcugagcau gccccaggug aacaucguga agaagaccga ggugcagacc ggcggcuuca     3360

gcaaggagag cauccugccc aagcggaaca gcgacaagcu gaucgcccgg aagaaggacu     3420

gggaccccaa gaaguacggc ggcuucgaca gccccaccgu ggccuacagc gugcuggugg     3480

uggccaaggu ggagaagggc aagagcaaga agcugaagag cgugaaggag cugcugggca     3540

ucaccaucau ggagcggagc agcuucgaga agaaccccau cgacuuccug gaggccaagg     3600

gcuacaagga ggugaagaag gaccugauca ucaagcugcc caaguacagc cuguucgagc     3660

uggagaacgg ccggaagcgg augcuggcca gcgccggcga gcugcagaag ggcaacgagc     3720

uggcccugcc cagcaaguac gugaacuucc uguaccuggc cagccacuac gagaagcuga     3780

agggcagccc cgaggacaac gagcagaagc agcuguucgu ggagcagcac aagcacuacc     3840

uggacgagau caucgagcag aucagcgagu ucagcaagcg ggugauccug gccgacgcca     3900

accuggacaa ggugcugagc gccuacaaca agcaccggga caagcccauc cgggagcagg     3960

ccgagaacau cauccaccug uucacccuga ccaaccuggg cgcccccgcc gccuucaagu     4020

acuucgacac caccaucgac cggaagcggu acaccagcac caaggaggug cuggacgcca     4080

cccugaucca ccagagcauc accggccugu acgagacccg gaucgaccug agccagcugg     4140

gcggcgacgg cggcggcagc cccaagaaga agcggaaggu gugacuagca ccagccucaa     4200

gaacacccga auggagucuc uaagcuacau aauaccaacu uacacuuuac aaaauguugu     4260

cccccaaaau guagccauuc guaucugcuc cuaauaaaaa gaaaguuucu ucacauucuc     4320

ucgagaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4380

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaucua g                         4421


<210>  179
<211>  4564
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with ORF encoding Cas9 with HiBiT tag,
       HSD 5 UTR and human ALB 3 UTR

<400>  179
gggtcccgca gtcggcgtcc agcggctctg cttgttcgtg tgtgtgtcgt tgcaggcctt       60

attcggatcc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtcagcgaaa gcgcaacacc ggaaagcgtc agcggatgga gactgttcaa     4260

gaagatcagc tagctagcca tcacatttaa aagcatctca gcctaccatg agaataagag     4320

aaagaaaatg aagatcaata gcttattcat ctctttttct ttttcgttgg tgtaaagcca     4380

acaccctgtc taaaaaacat aaatttcttt aatcattttg cctcttttct ctgtgcttca     4440

attaataaaa aatggaaaga acctcgagaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4500

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaat     4560

ctag                                                                  4564


<210>  180
<211>  4544
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with ORF encoding Cas9 with HiBiT tag,
       CMV-1 5 UTR and human ALB 3 UTR

<400>  180
gggcagatcg cctggagacg ccatccacgc tgttttgacc tccatcgcca ccatggacaa       60

gaagtacagc atcggactgg acatcggaac aaacagcgtc ggatgggcag tcatcacaga      120

cgaatacaag gtcccgagca agaagttcaa ggtcctggga aacacagaca gacacagcat      180

caagaagaac ctgatcggag cactgctgtt cgacagcgga gaaacagcag aagcaacaag      240

actgaagaga acagcaagaa gaagatacac aagaagaaag aacagaatct gctacctgca      300

ggaaatcttc agcaacgaaa tggcaaaggt cgacgacagc ttcttccaca gactggaaga      360

aagcttcctg gtcgaagaag acaagaagca cgaaagacac ccgatcttcg gaaacatcgt      420

cgacgaagtc gcataccacg aaaagtaccc gacaatctac cacctgagaa agaagctggt      480

cgacagcaca gacaaggcag acctgagact gatctacctg gcactggcac acatgatcaa      540

gttcagagga cacttcctga tcgaaggaga cctgaacccg gacaacagcg acgtcgacaa      600

gctgttcatc cagctggtcc agacatacaa ccagctgttc gaagaaaacc cgatcaacgc      660

aagcggagtc gacgcaaagg caatcctgag cgcaagactg agcaagagca gaagactgga      720

aaacctgatc gcacagctgc cgggagaaaa gaagaacgga ctgttcggaa acctgatcgc      780

actgagcctg ggactgacac cgaacttcaa gagcaacttc gacctggcag aagacgcaaa      840

gctgcagctg agcaaggaca catacgacga cgacctggac aacctgctgg cacagatcgg      900

agaccagtac gcagacctgt tcctggcagc aaagaacctg agcgacgcaa tcctgctgag      960

cgacatcctg agagtcaaca cagaaatcac aaaggcaccg ctgagcgcaa gcatgatcaa     1020

gagatacgac gaacaccacc aggacctgac actgctgaag gcactggtca gacagcagct     1080

gccggaaaag tacaaggaaa tcttcttcga ccagagcaag aacggatacg caggatacat     1140

cgacggagga gcaagccagg aagaattcta caagttcatc aagccgatcc tggaaaagat     1200

ggacggaaca gaagaactgc tggtcaagct gaacagagaa gacctgctga gaaagcagag     1260

aacattcgac aacggaagca tcccgcacca gatccacctg ggagaactgc acgcaatcct     1320

gagaagacag gaagacttct acccgttcct gaaggacaac agagaaaaga tcgaaaagat     1380

cctgacattc agaatcccgt actacgtcgg accgctggca agaggaaaca gcagattcgc     1440

atggatgaca agaaagagcg aagaaacaat cacaccgtgg aacttcgaag aagtcgtcga     1500

caagggagca agcgcacaga gcttcatcga aagaatgaca aacttcgaca agaacctgcc     1560

gaacgaaaag gtcctgccga agcacagcct gctgtacgaa tacttcacag tctacaacga     1620

actgacaaag gtcaagtacg tcacagaagg aatgagaaag ccggcattcc tgagcggaga     1680

acagaagaag gcaatcgtcg acctgctgtt caagacaaac agaaaggtca cagtcaagca     1740

gctgaaggaa gactacttca agaagatcga atgcttcgac agcgtcgaaa tcagcggagt     1800

cgaagacaga ttcaacgcaa gcctgggaac ataccacgac ctgctgaaga tcatcaagga     1860

caaggacttc ctggacaacg aagaaaacga agacatcctg gaagacatcg tcctgacact     1920

gacactgttc gaagacagag aaatgatcga agaaagactg aagacatacg cacacctgtt     1980

cgacgacaag gtcatgaagc agctgaagag aagaagatac acaggatggg gaagactgag     2040

cagaaagctg atcaacggaa tcagagacaa gcagagcgga aagacaatcc tggacttcct     2100

gaagagcgac ggattcgcaa acagaaactt catgcagctg atccacgacg acagcctgac     2160

attcaaggaa gacatccaga aggcacaggt cagcggacag ggagacagcc tgcacgaaca     2220

catcgcaaac ctggcaggaa gcccggcaat caagaaggga atcctgcaga cagtcaaggt     2280

cgtcgacgaa ctggtcaagg tcatgggaag acacaagccg gaaaacatcg tcatcgaaat     2340

ggcaagagaa aaccagacaa cacagaaggg acagaagaac agcagagaaa gaatgaagag     2400

aatcgaagaa ggaatcaagg aactgggaag ccagatcctg aaggaacacc cggtcgaaaa     2460

cacacagctg cagaacgaaa agctgtacct gtactacctg cagaacggaa gagacatgta     2520

cgtcgaccag gaactggaca tcaacagact gagcgactac gacgtcgacc acatcgtccc     2580

gcagagcttc ctgaaggacg acagcatcga caacaaggtc ctgacaagaa gcgacaagaa     2640

cagaggaaag agcgacaacg tcccgagcga agaagtcgtc aagaagatga agaactactg     2700

gagacagctg ctgaacgcaa agctgatcac acagagaaag ttcgacaacc tgacaaaggc     2760

agagagagga ggactgagcg aactggacaa ggcaggattc atcaagagac agctggtcga     2820

aacaagacag atcacaaagc acgtcgcaca gatcctggac agcagaatga acacaaagta     2880

cgacgaaaac gacaagctga tcagagaagt caaggtcatc acactgaaga gcaagctggt     2940

cagcgacttc agaaaggact tccagttcta caaggtcaga gaaatcaaca actaccacca     3000

cgcacacgac gcatacctga acgcagtcgt cggaacagca ctgatcaaga agtacccgaa     3060

gctggaaagc gaattcgtct acggagacta caaggtctac gacgtcagaa agatgatcgc     3120

aaagagcgaa caggaaatcg gaaaggcaac agcaaagtac ttcttctaca gcaacatcat     3180

gaacttcttc aagacagaaa tcacactggc aaacggagaa atcagaaaga gaccgctgat     3240

cgaaacaaac ggagaaacag gagaaatcgt ctgggacaag ggaagagact tcgcaacagt     3300

cagaaaggtc ctgagcatgc cgcaggtcaa catcgtcaag aagacagaag tccagacagg     3360

aggattcagc aaggaaagca tcctgccgaa gagaaacagc gacaagctga tcgcaagaaa     3420

gaaggactgg gacccgaaga agtacggagg attcgacagc ccgacagtcg catacagcgt     3480

cctggtcgtc gcaaaggtcg aaaagggaaa gagcaagaag ctgaagagcg tcaaggaact     3540

gctgggaatc acaatcatgg aaagaagcag cttcgaaaag aacccgatcg acttcctgga     3600

agcaaaggga tacaaggaag tcaagaagga cctgatcatc aagctgccga agtacagcct     3660

gttcgaactg gaaaacggaa gaaagagaat gctggcaagc gcaggagaac tgcagaaggg     3720

aaacgaactg gcactgccga gcaagtacgt caacttcctg tacctggcaa gccactacga     3780

aaagctgaag ggaagcccgg aagacaacga acagaagcag ctgttcgtcg aacagcacaa     3840

gcactacctg gacgaaatca tcgaacagat cagcgaattc agcaagagag tcatcctggc     3900

agacgcaaac ctggacaagg tcctgagcgc atacaacaag cacagagaca agccgatcag     3960

agaacaggca gaaaacatca tccacctgtt cacactgaca aacctgggag caccggcagc     4020

attcaagtac ttcgacacaa caatcgacag aaagagatac acaagcacaa aggaagtcct     4080

ggacgcaaca ctgatccacc agagcatcac aggactgtac gaaacaagaa tcgacctgag     4140

ccagctggga ggagacggag gaggaagccc gaagaagaag agaaaggtca gcgaaagcgc     4200

aacaccggaa agcgtcagcg gatggagact gttcaagaag atcagctagc tagccatcac     4260

atttaaaagc atctcagcct accatgagaa taagagaaag aaaatgaaga tcaatagctt     4320

attcatctct ttttcttttt cgttggtgta aagccaacac cctgtctaaa aaacataaat     4380

ttctttaatc attttgcctc ttttctctgt gcttcaatta ataaaaaatg gaaagaacct     4440

cgagaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4500

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa                      4544


<210>  181
<211>  4544
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with ORF encoding Cas9 with HiBiT tag,
       CMV-2 5 UTR and human ALB 3 UTR

<400>  181
gggagaagac accgggaccg atccagcctc cgcggccggg aacggcgcca ccatggacaa       60

gaagtacagc atcggactgg acatcggaac aaacagcgtc ggatgggcag tcatcacaga      120

cgaatacaag gtcccgagca agaagttcaa ggtcctggga aacacagaca gacacagcat      180

caagaagaac ctgatcggag cactgctgtt cgacagcgga gaaacagcag aagcaacaag      240

actgaagaga acagcaagaa gaagatacac aagaagaaag aacagaatct gctacctgca      300

ggaaatcttc agcaacgaaa tggcaaaggt cgacgacagc ttcttccaca gactggaaga      360

aagcttcctg gtcgaagaag acaagaagca cgaaagacac ccgatcttcg gaaacatcgt      420

cgacgaagtc gcataccacg aaaagtaccc gacaatctac cacctgagaa agaagctggt      480

cgacagcaca gacaaggcag acctgagact gatctacctg gcactggcac acatgatcaa      540

gttcagagga cacttcctga tcgaaggaga cctgaacccg gacaacagcg acgtcgacaa      600

gctgttcatc cagctggtcc agacatacaa ccagctgttc gaagaaaacc cgatcaacgc      660

aagcggagtc gacgcaaagg caatcctgag cgcaagactg agcaagagca gaagactgga      720

aaacctgatc gcacagctgc cgggagaaaa gaagaacgga ctgttcggaa acctgatcgc      780

actgagcctg ggactgacac cgaacttcaa gagcaacttc gacctggcag aagacgcaaa      840

gctgcagctg agcaaggaca catacgacga cgacctggac aacctgctgg cacagatcgg      900

agaccagtac gcagacctgt tcctggcagc aaagaacctg agcgacgcaa tcctgctgag      960

cgacatcctg agagtcaaca cagaaatcac aaaggcaccg ctgagcgcaa gcatgatcaa     1020

gagatacgac gaacaccacc aggacctgac actgctgaag gcactggtca gacagcagct     1080

gccggaaaag tacaaggaaa tcttcttcga ccagagcaag aacggatacg caggatacat     1140

cgacggagga gcaagccagg aagaattcta caagttcatc aagccgatcc tggaaaagat     1200

ggacggaaca gaagaactgc tggtcaagct gaacagagaa gacctgctga gaaagcagag     1260

aacattcgac aacggaagca tcccgcacca gatccacctg ggagaactgc acgcaatcct     1320

gagaagacag gaagacttct acccgttcct gaaggacaac agagaaaaga tcgaaaagat     1380

cctgacattc agaatcccgt actacgtcgg accgctggca agaggaaaca gcagattcgc     1440

atggatgaca agaaagagcg aagaaacaat cacaccgtgg aacttcgaag aagtcgtcga     1500

caagggagca agcgcacaga gcttcatcga aagaatgaca aacttcgaca agaacctgcc     1560

gaacgaaaag gtcctgccga agcacagcct gctgtacgaa tacttcacag tctacaacga     1620

actgacaaag gtcaagtacg tcacagaagg aatgagaaag ccggcattcc tgagcggaga     1680

acagaagaag gcaatcgtcg acctgctgtt caagacaaac agaaaggtca cagtcaagca     1740

gctgaaggaa gactacttca agaagatcga atgcttcgac agcgtcgaaa tcagcggagt     1800

cgaagacaga ttcaacgcaa gcctgggaac ataccacgac ctgctgaaga tcatcaagga     1860

caaggacttc ctggacaacg aagaaaacga agacatcctg gaagacatcg tcctgacact     1920

gacactgttc gaagacagag aaatgatcga agaaagactg aagacatacg cacacctgtt     1980

cgacgacaag gtcatgaagc agctgaagag aagaagatac acaggatggg gaagactgag     2040

cagaaagctg atcaacggaa tcagagacaa gcagagcgga aagacaatcc tggacttcct     2100

gaagagcgac ggattcgcaa acagaaactt catgcagctg atccacgacg acagcctgac     2160

attcaaggaa gacatccaga aggcacaggt cagcggacag ggagacagcc tgcacgaaca     2220

catcgcaaac ctggcaggaa gcccggcaat caagaaggga atcctgcaga cagtcaaggt     2280

cgtcgacgaa ctggtcaagg tcatgggaag acacaagccg gaaaacatcg tcatcgaaat     2340

ggcaagagaa aaccagacaa cacagaaggg acagaagaac agcagagaaa gaatgaagag     2400

aatcgaagaa ggaatcaagg aactgggaag ccagatcctg aaggaacacc cggtcgaaaa     2460

cacacagctg cagaacgaaa agctgtacct gtactacctg cagaacggaa gagacatgta     2520

cgtcgaccag gaactggaca tcaacagact gagcgactac gacgtcgacc acatcgtccc     2580

gcagagcttc ctgaaggacg acagcatcga caacaaggtc ctgacaagaa gcgacaagaa     2640

cagaggaaag agcgacaacg tcccgagcga agaagtcgtc aagaagatga agaactactg     2700

gagacagctg ctgaacgcaa agctgatcac acagagaaag ttcgacaacc tgacaaaggc     2760

agagagagga ggactgagcg aactggacaa ggcaggattc atcaagagac agctggtcga     2820

aacaagacag atcacaaagc acgtcgcaca gatcctggac agcagaatga acacaaagta     2880

cgacgaaaac gacaagctga tcagagaagt caaggtcatc acactgaaga gcaagctggt     2940

cagcgacttc agaaaggact tccagttcta caaggtcaga gaaatcaaca actaccacca     3000

cgcacacgac gcatacctga acgcagtcgt cggaacagca ctgatcaaga agtacccgaa     3060

gctggaaagc gaattcgtct acggagacta caaggtctac gacgtcagaa agatgatcgc     3120

aaagagcgaa caggaaatcg gaaaggcaac agcaaagtac ttcttctaca gcaacatcat     3180

gaacttcttc aagacagaaa tcacactggc aaacggagaa atcagaaaga gaccgctgat     3240

cgaaacaaac ggagaaacag gagaaatcgt ctgggacaag ggaagagact tcgcaacagt     3300

cagaaaggtc ctgagcatgc cgcaggtcaa catcgtcaag aagacagaag tccagacagg     3360

aggattcagc aaggaaagca tcctgccgaa gagaaacagc gacaagctga tcgcaagaaa     3420

gaaggactgg gacccgaaga agtacggagg attcgacagc ccgacagtcg catacagcgt     3480

cctggtcgtc gcaaaggtcg aaaagggaaa gagcaagaag ctgaagagcg tcaaggaact     3540

gctgggaatc acaatcatgg aaagaagcag cttcgaaaag aacccgatcg acttcctgga     3600

agcaaaggga tacaaggaag tcaagaagga cctgatcatc aagctgccga agtacagcct     3660

gttcgaactg gaaaacggaa gaaagagaat gctggcaagc gcaggagaac tgcagaaggg     3720

aaacgaactg gcactgccga gcaagtacgt caacttcctg tacctggcaa gccactacga     3780

aaagctgaag ggaagcccgg aagacaacga acagaagcag ctgttcgtcg aacagcacaa     3840

gcactacctg gacgaaatca tcgaacagat cagcgaattc agcaagagag tcatcctggc     3900

agacgcaaac ctggacaagg tcctgagcgc atacaacaag cacagagaca agccgatcag     3960

agaacaggca gaaaacatca tccacctgtt cacactgaca aacctgggag caccggcagc     4020

attcaagtac ttcgacacaa caatcgacag aaagagatac acaagcacaa aggaagtcct     4080

ggacgcaaca ctgatccacc agagcatcac aggactgtac gaaacaagaa tcgacctgag     4140

ccagctggga ggagacggag gaggaagccc gaagaagaag agaaaggtca gcgaaagcgc     4200

aacaccggaa agcgtcagcg gatggagact gttcaagaag atcagctagc tagccatcac     4260

atttaaaagc atctcagcct accatgagaa taagagaaag aaaatgaaga tcaatagctt     4320

attcatctct ttttcttttt cgttggtgta aagccaacac cctgtctaaa aaacataaat     4380

ttctttaatc attttgcctc ttttctctgt gcttcaatta ataaaaaatg gaaagaacct     4440

cgagaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4500

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa                      4544


<210>  182
<211>  4544
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with ORF encoding Cas9 with HiBiT tag,
       CMV-3 5 UTR and human ALB 3 UTR

<400>  182
gggtgcattg gaacgcggat tccccgtgcc aagagtgact caccgcgcca ccatggacaa       60

gaagtacagc atcggactgg acatcggaac aaacagcgtc ggatgggcag tcatcacaga      120

cgaatacaag gtcccgagca agaagttcaa ggtcctggga aacacagaca gacacagcat      180

caagaagaac ctgatcggag cactgctgtt cgacagcgga gaaacagcag aagcaacaag      240

actgaagaga acagcaagaa gaagatacac aagaagaaag aacagaatct gctacctgca      300

ggaaatcttc agcaacgaaa tggcaaaggt cgacgacagc ttcttccaca gactggaaga      360

aagcttcctg gtcgaagaag acaagaagca cgaaagacac ccgatcttcg gaaacatcgt      420

cgacgaagtc gcataccacg aaaagtaccc gacaatctac cacctgagaa agaagctggt      480

cgacagcaca gacaaggcag acctgagact gatctacctg gcactggcac acatgatcaa      540

gttcagagga cacttcctga tcgaaggaga cctgaacccg gacaacagcg acgtcgacaa      600

gctgttcatc cagctggtcc agacatacaa ccagctgttc gaagaaaacc cgatcaacgc      660

aagcggagtc gacgcaaagg caatcctgag cgcaagactg agcaagagca gaagactgga      720

aaacctgatc gcacagctgc cgggagaaaa gaagaacgga ctgttcggaa acctgatcgc      780

actgagcctg ggactgacac cgaacttcaa gagcaacttc gacctggcag aagacgcaaa      840

gctgcagctg agcaaggaca catacgacga cgacctggac aacctgctgg cacagatcgg      900

agaccagtac gcagacctgt tcctggcagc aaagaacctg agcgacgcaa tcctgctgag      960

cgacatcctg agagtcaaca cagaaatcac aaaggcaccg ctgagcgcaa gcatgatcaa     1020

gagatacgac gaacaccacc aggacctgac actgctgaag gcactggtca gacagcagct     1080

gccggaaaag tacaaggaaa tcttcttcga ccagagcaag aacggatacg caggatacat     1140

cgacggagga gcaagccagg aagaattcta caagttcatc aagccgatcc tggaaaagat     1200

ggacggaaca gaagaactgc tggtcaagct gaacagagaa gacctgctga gaaagcagag     1260

aacattcgac aacggaagca tcccgcacca gatccacctg ggagaactgc acgcaatcct     1320

gagaagacag gaagacttct acccgttcct gaaggacaac agagaaaaga tcgaaaagat     1380

cctgacattc agaatcccgt actacgtcgg accgctggca agaggaaaca gcagattcgc     1440

atggatgaca agaaagagcg aagaaacaat cacaccgtgg aacttcgaag aagtcgtcga     1500

caagggagca agcgcacaga gcttcatcga aagaatgaca aacttcgaca agaacctgcc     1560

gaacgaaaag gtcctgccga agcacagcct gctgtacgaa tacttcacag tctacaacga     1620

actgacaaag gtcaagtacg tcacagaagg aatgagaaag ccggcattcc tgagcggaga     1680

acagaagaag gcaatcgtcg acctgctgtt caagacaaac agaaaggtca cagtcaagca     1740

gctgaaggaa gactacttca agaagatcga atgcttcgac agcgtcgaaa tcagcggagt     1800

cgaagacaga ttcaacgcaa gcctgggaac ataccacgac ctgctgaaga tcatcaagga     1860

caaggacttc ctggacaacg aagaaaacga agacatcctg gaagacatcg tcctgacact     1920

gacactgttc gaagacagag aaatgatcga agaaagactg aagacatacg cacacctgtt     1980

cgacgacaag gtcatgaagc agctgaagag aagaagatac acaggatggg gaagactgag     2040

cagaaagctg atcaacggaa tcagagacaa gcagagcgga aagacaatcc tggacttcct     2100

gaagagcgac ggattcgcaa acagaaactt catgcagctg atccacgacg acagcctgac     2160

attcaaggaa gacatccaga aggcacaggt cagcggacag ggagacagcc tgcacgaaca     2220

catcgcaaac ctggcaggaa gcccggcaat caagaaggga atcctgcaga cagtcaaggt     2280

cgtcgacgaa ctggtcaagg tcatgggaag acacaagccg gaaaacatcg tcatcgaaat     2340

ggcaagagaa aaccagacaa cacagaaggg acagaagaac agcagagaaa gaatgaagag     2400

aatcgaagaa ggaatcaagg aactgggaag ccagatcctg aaggaacacc cggtcgaaaa     2460

cacacagctg cagaacgaaa agctgtacct gtactacctg cagaacggaa gagacatgta     2520

cgtcgaccag gaactggaca tcaacagact gagcgactac gacgtcgacc acatcgtccc     2580

gcagagcttc ctgaaggacg acagcatcga caacaaggtc ctgacaagaa gcgacaagaa     2640

cagaggaaag agcgacaacg tcccgagcga agaagtcgtc aagaagatga agaactactg     2700

gagacagctg ctgaacgcaa agctgatcac acagagaaag ttcgacaacc tgacaaaggc     2760

agagagagga ggactgagcg aactggacaa ggcaggattc atcaagagac agctggtcga     2820

aacaagacag atcacaaagc acgtcgcaca gatcctggac agcagaatga acacaaagta     2880

cgacgaaaac gacaagctga tcagagaagt caaggtcatc acactgaaga gcaagctggt     2940

cagcgacttc agaaaggact tccagttcta caaggtcaga gaaatcaaca actaccacca     3000

cgcacacgac gcatacctga acgcagtcgt cggaacagca ctgatcaaga agtacccgaa     3060

gctggaaagc gaattcgtct acggagacta caaggtctac gacgtcagaa agatgatcgc     3120

aaagagcgaa caggaaatcg gaaaggcaac agcaaagtac ttcttctaca gcaacatcat     3180

gaacttcttc aagacagaaa tcacactggc aaacggagaa atcagaaaga gaccgctgat     3240

cgaaacaaac ggagaaacag gagaaatcgt ctgggacaag ggaagagact tcgcaacagt     3300

cagaaaggtc ctgagcatgc cgcaggtcaa catcgtcaag aagacagaag tccagacagg     3360

aggattcagc aaggaaagca tcctgccgaa gagaaacagc gacaagctga tcgcaagaaa     3420

gaaggactgg gacccgaaga agtacggagg attcgacagc ccgacagtcg catacagcgt     3480

cctggtcgtc gcaaaggtcg aaaagggaaa gagcaagaag ctgaagagcg tcaaggaact     3540

gctgggaatc acaatcatgg aaagaagcag cttcgaaaag aacccgatcg acttcctgga     3600

agcaaaggga tacaaggaag tcaagaagga cctgatcatc aagctgccga agtacagcct     3660

gttcgaactg gaaaacggaa gaaagagaat gctggcaagc gcaggagaac tgcagaaggg     3720

aaacgaactg gcactgccga gcaagtacgt caacttcctg tacctggcaa gccactacga     3780

aaagctgaag ggaagcccgg aagacaacga acagaagcag ctgttcgtcg aacagcacaa     3840

gcactacctg gacgaaatca tcgaacagat cagcgaattc agcaagagag tcatcctggc     3900

agacgcaaac ctggacaagg tcctgagcgc atacaacaag cacagagaca agccgatcag     3960

agaacaggca gaaaacatca tccacctgtt cacactgaca aacctgggag caccggcagc     4020

attcaagtac ttcgacacaa caatcgacag aaagagatac acaagcacaa aggaagtcct     4080

ggacgcaaca ctgatccacc agagcatcac aggactgtac gaaacaagaa tcgacctgag     4140

ccagctggga ggagacggag gaggaagccc gaagaagaag agaaaggtca gcgaaagcgc     4200

aacaccggaa agcgtcagcg gatggagact gttcaagaag atcagctagc tagccatcac     4260

atttaaaagc atctcagcct accatgagaa taagagaaag aaaatgaaga tcaatagctt     4320

attcatctct ttttcttttt cgttggtgta aagccaacac cctgtctaaa aaacataaat     4380

ttctttaatc attttgcctc ttttctctgt gcttcaatta ataaaaaatg gaaagaacct     4440

cgagaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4500

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa                      4544


<210>  183
<211>  4568
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with ORF encoding Cas9 with HiBiT tag,
       HBA 5 UTR and human ALB 3 UTR

<400>  183
gggcataaac cctggcgcgc tcgcggcccg gcactcttct ggtccccaca gactcagaga       60

gaacccaccc gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag      120

cgtcggatgg gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct      180

gggaaacaca gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag      240

cggagaaaca gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag      300

aaagaacaga atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga      360

cagcttcttc cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag      420

acacccgatc ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat      480

ctaccacctg agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta      540

cctggcactg gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa      600

cccggacaac agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct      660

gttcgaagaa aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag      720

actgagcaag agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa      780

cggactgttc ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa      840

cttcgacctg gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct      900

ggacaacctg ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa      960

cctgagcgac gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc     1020

accgctgagc gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct     1080

gaaggcactg gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag     1140

caagaacgga tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt     1200

catcaagccg atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag     1260

agaagacctg ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca     1320

cctgggagaa ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga     1380

caacagagaa aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct     1440

ggcaagagga aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc     1500

gtggaacttc gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat     1560

gacaaacttc gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta     1620

cgaatacttc acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag     1680

aaagccggca ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac     1740

aaacagaaag gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt     1800

cgacagcgtc gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca     1860

cgacctgctg aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat     1920

cctggaagac atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag     1980

actgaagaca tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag     2040

atacacagga tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag     2100

cggaaagaca atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca     2160

gctgatccac gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg     2220

acagggagac agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa     2280

gggaatcctg cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa     2340

gccggaaaac atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa     2400

gaacagcaga gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat     2460

cctgaaggaa cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta     2520

cctgcagaac ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga     2580

ctacgacgtc gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa     2640

ggtcctgaca agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt     2700

cgtcaagaag atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag     2760

aaagttcgac aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg     2820

attcatcaag agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct     2880

ggacagcaga atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt     2940

catcacactg aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt     3000

cagagaaatc aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac     3060

agcactgatc aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt     3120

ctacgacgtc agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa     3180

gtacttcttc tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg     3240

agaaatcaga aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga     3300

caagggaaga gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt     3360

caagaagaca gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa     3420

cagcgacaag ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga     3480

cagcccgaca gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa     3540

gaagctgaag agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga     3600

aaagaacccg atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat     3660

catcaagctg ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc     3720

aagcgcagga gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt     3780

cctgtacctg gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa     3840

gcagctgttc gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga     3900

attcagcaag agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa     3960

caagcacaga gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact     4020

gacaaacctg ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag     4080

atacacaagc acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact     4140

gtacgaaaca agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa     4200

gaagagaaag gtcagcgaaa gcgcaacacc ggaaagcgtc agcggatgga gactgttcaa     4260

gaagatcagc tagctagcca tcacatttaa aagcatctca gcctaccatg agaataagag     4320

aaagaaaatg aagatcaata gcttattcat ctctttttct ttttcgttgg tgtaaagcca     4380

acaccctgtc taaaaaacat aaatttcttt aatcattttg cctcttttct ctgtgcttca     4440

attaataaaa aatggaaaga acctcgagaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4500

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4560

aaaaaaaa                                                              4568


<210>  184
<211>  4558
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with ORF encoding Cas9 with HiBiT tag,
       HBB 5 UTR and human ALB 3 UTR

<400>  184
gggacatttg cttctgacac aactgtgttc actagcaacc tcaaacagac accggatctc       60

gccaccatgg acaagaagta cagcatcgga ctggacatcg gaacaaacag cgtcggatgg      120

gcagtcatca cagacgaata caaggtcccg agcaagaagt tcaaggtcct gggaaacaca      180

gacagacaca gcatcaagaa gaacctgatc ggagcactgc tgttcgacag cggagaaaca      240

gcagaagcaa caagactgaa gagaacagca agaagaagat acacaagaag aaagaacaga      300

atctgctacc tgcaggaaat cttcagcaac gaaatggcaa aggtcgacga cagcttcttc      360

cacagactgg aagaaagctt cctggtcgaa gaagacaaga agcacgaaag acacccgatc      420

ttcggaaaca tcgtcgacga agtcgcatac cacgaaaagt acccgacaat ctaccacctg      480

agaaagaagc tggtcgacag cacagacaag gcagacctga gactgatcta cctggcactg      540

gcacacatga tcaagttcag aggacacttc ctgatcgaag gagacctgaa cccggacaac      600

agcgacgtcg acaagctgtt catccagctg gtccagacat acaaccagct gttcgaagaa      660

aacccgatca acgcaagcgg agtcgacgca aaggcaatcc tgagcgcaag actgagcaag      720

agcagaagac tggaaaacct gatcgcacag ctgccgggag aaaagaagaa cggactgttc      780

ggaaacctga tcgcactgag cctgggactg acaccgaact tcaagagcaa cttcgacctg      840

gcagaagacg caaagctgca gctgagcaag gacacatacg acgacgacct ggacaacctg      900

ctggcacaga tcggagacca gtacgcagac ctgttcctgg cagcaaagaa cctgagcgac      960

gcaatcctgc tgagcgacat cctgagagtc aacacagaaa tcacaaaggc accgctgagc     1020

gcaagcatga tcaagagata cgacgaacac caccaggacc tgacactgct gaaggcactg     1080

gtcagacagc agctgccgga aaagtacaag gaaatcttct tcgaccagag caagaacgga     1140

tacgcaggat acatcgacgg aggagcaagc caggaagaat tctacaagtt catcaagccg     1200

atcctggaaa agatggacgg aacagaagaa ctgctggtca agctgaacag agaagacctg     1260

ctgagaaagc agagaacatt cgacaacgga agcatcccgc accagatcca cctgggagaa     1320

ctgcacgcaa tcctgagaag acaggaagac ttctacccgt tcctgaagga caacagagaa     1380

aagatcgaaa agatcctgac attcagaatc ccgtactacg tcggaccgct ggcaagagga     1440

aacagcagat tcgcatggat gacaagaaag agcgaagaaa caatcacacc gtggaacttc     1500

gaagaagtcg tcgacaaggg agcaagcgca cagagcttca tcgaaagaat gacaaacttc     1560

gacaagaacc tgccgaacga aaaggtcctg ccgaagcaca gcctgctgta cgaatacttc     1620

acagtctaca acgaactgac aaaggtcaag tacgtcacag aaggaatgag aaagccggca     1680

ttcctgagcg gagaacagaa gaaggcaatc gtcgacctgc tgttcaagac aaacagaaag     1740

gtcacagtca agcagctgaa ggaagactac ttcaagaaga tcgaatgctt cgacagcgtc     1800

gaaatcagcg gagtcgaaga cagattcaac gcaagcctgg gaacatacca cgacctgctg     1860

aagatcatca aggacaagga cttcctggac aacgaagaaa acgaagacat cctggaagac     1920

atcgtcctga cactgacact gttcgaagac agagaaatga tcgaagaaag actgaagaca     1980

tacgcacacc tgttcgacga caaggtcatg aagcagctga agagaagaag atacacagga     2040

tggggaagac tgagcagaaa gctgatcaac ggaatcagag acaagcagag cggaaagaca     2100

atcctggact tcctgaagag cgacggattc gcaaacagaa acttcatgca gctgatccac     2160

gacgacagcc tgacattcaa ggaagacatc cagaaggcac aggtcagcgg acagggagac     2220

agcctgcacg aacacatcgc aaacctggca ggaagcccgg caatcaagaa gggaatcctg     2280

cagacagtca aggtcgtcga cgaactggtc aaggtcatgg gaagacacaa gccggaaaac     2340

atcgtcatcg aaatggcaag agaaaaccag acaacacaga agggacagaa gaacagcaga     2400

gaaagaatga agagaatcga agaaggaatc aaggaactgg gaagccagat cctgaaggaa     2460

cacccggtcg aaaacacaca gctgcagaac gaaaagctgt acctgtacta cctgcagaac     2520

ggaagagaca tgtacgtcga ccaggaactg gacatcaaca gactgagcga ctacgacgtc     2580

gaccacatcg tcccgcagag cttcctgaag gacgacagca tcgacaacaa ggtcctgaca     2640

agaagcgaca agaacagagg aaagagcgac aacgtcccga gcgaagaagt cgtcaagaag     2700

atgaagaact actggagaca gctgctgaac gcaaagctga tcacacagag aaagttcgac     2760

aacctgacaa aggcagagag aggaggactg agcgaactgg acaaggcagg attcatcaag     2820

agacagctgg tcgaaacaag acagatcaca aagcacgtcg cacagatcct ggacagcaga     2880

atgaacacaa agtacgacga aaacgacaag ctgatcagag aagtcaaggt catcacactg     2940

aagagcaagc tggtcagcga cttcagaaag gacttccagt tctacaaggt cagagaaatc     3000

aacaactacc accacgcaca cgacgcatac ctgaacgcag tcgtcggaac agcactgatc     3060

aagaagtacc cgaagctgga aagcgaattc gtctacggag actacaaggt ctacgacgtc     3120

agaaagatga tcgcaaagag cgaacaggaa atcggaaagg caacagcaaa gtacttcttc     3180

tacagcaaca tcatgaactt cttcaagaca gaaatcacac tggcaaacgg agaaatcaga     3240

aagagaccgc tgatcgaaac aaacggagaa acaggagaaa tcgtctggga caagggaaga     3300

gacttcgcaa cagtcagaaa ggtcctgagc atgccgcagg tcaacatcgt caagaagaca     3360

gaagtccaga caggaggatt cagcaaggaa agcatcctgc cgaagagaaa cagcgacaag     3420

ctgatcgcaa gaaagaagga ctgggacccg aagaagtacg gaggattcga cagcccgaca     3480

gtcgcataca gcgtcctggt cgtcgcaaag gtcgaaaagg gaaagagcaa gaagctgaag     3540

agcgtcaagg aactgctggg aatcacaatc atggaaagaa gcagcttcga aaagaacccg     3600

atcgacttcc tggaagcaaa gggatacaag gaagtcaaga aggacctgat catcaagctg     3660

ccgaagtaca gcctgttcga actggaaaac ggaagaaaga gaatgctggc aagcgcagga     3720

gaactgcaga agggaaacga actggcactg ccgagcaagt acgtcaactt cctgtacctg     3780

gcaagccact acgaaaagct gaagggaagc ccggaagaca acgaacagaa gcagctgttc     3840

gtcgaacagc acaagcacta cctggacgaa atcatcgaac agatcagcga attcagcaag     3900

agagtcatcc tggcagacgc aaacctggac aaggtcctga gcgcatacaa caagcacaga     3960

gacaagccga tcagagaaca ggcagaaaac atcatccacc tgttcacact gacaaacctg     4020

ggagcaccgg cagcattcaa gtacttcgac acaacaatcg acagaaagag atacacaagc     4080

acaaaggaag tcctggacgc aacactgatc caccagagca tcacaggact gtacgaaaca     4140

agaatcgacc tgagccagct gggaggagac ggaggaggaa gcccgaagaa gaagagaaag     4200

gtcagcgaaa gcgcaacacc ggaaagcgtc agcggatgga gactgttcaa gaagatcagc     4260

tagctagcca tcacatttaa aagcatctca gcctaccatg agaataagag aaagaaaatg     4320

aagatcaata gcttattcat ctctttttct ttttcgttgg tgtaaagcca acaccctgtc     4380

taaaaaacat aaatttcttt aatcattttg cctcttttct ctgtgcttca attaataaaa     4440

aatggaaaga acctcgagaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4500

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaa       4558


<210>  185
<211>  4537
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: mRNA transcript with ORF encoding Cas9 with HiBiT tag,
       XBG 5 UTR and human ALB 3 UTR

<400>  185
gggaagctca gaataaacgc tcaactttgg ccggatctcg ccaccatgga caagaagtac       60

agcatcggac tggacatcgg aacaaacagc gtcggatggg cagtcatcac agacgaatac      120

aaggtcccga gcaagaagtt caaggtcctg ggaaacacag acagacacag catcaagaag      180

aacctgatcg gagcactgct gttcgacagc ggagaaacag cagaagcaac aagactgaag      240

agaacagcaa gaagaagata cacaagaaga aagaacagaa tctgctacct gcaggaaatc      300

ttcagcaacg aaatggcaaa ggtcgacgac agcttcttcc acagactgga agaaagcttc      360

ctggtcgaag aagacaagaa gcacgaaaga cacccgatct tcggaaacat cgtcgacgaa      420

gtcgcatacc acgaaaagta cccgacaatc taccacctga gaaagaagct ggtcgacagc      480

acagacaagg cagacctgag actgatctac ctggcactgg cacacatgat caagttcaga      540

ggacacttcc tgatcgaagg agacctgaac ccggacaaca gcgacgtcga caagctgttc      600

atccagctgg tccagacata caaccagctg ttcgaagaaa acccgatcaa cgcaagcgga      660

gtcgacgcaa aggcaatcct gagcgcaaga ctgagcaaga gcagaagact ggaaaacctg      720

atcgcacagc tgccgggaga aaagaagaac ggactgttcg gaaacctgat cgcactgagc      780

ctgggactga caccgaactt caagagcaac ttcgacctgg cagaagacgc aaagctgcag      840

ctgagcaagg acacatacga cgacgacctg gacaacctgc tggcacagat cggagaccag      900

tacgcagacc tgttcctggc agcaaagaac ctgagcgacg caatcctgct gagcgacatc      960

ctgagagtca acacagaaat cacaaaggca ccgctgagcg caagcatgat caagagatac     1020

gacgaacacc accaggacct gacactgctg aaggcactgg tcagacagca gctgccggaa     1080

aagtacaagg aaatcttctt cgaccagagc aagaacggat acgcaggata catcgacgga     1140

ggagcaagcc aggaagaatt ctacaagttc atcaagccga tcctggaaaa gatggacgga     1200

acagaagaac tgctggtcaa gctgaacaga gaagacctgc tgagaaagca gagaacattc     1260

gacaacggaa gcatcccgca ccagatccac ctgggagaac tgcacgcaat cctgagaaga     1320

caggaagact tctacccgtt cctgaaggac aacagagaaa agatcgaaaa gatcctgaca     1380

ttcagaatcc cgtactacgt cggaccgctg gcaagaggaa acagcagatt cgcatggatg     1440

acaagaaaga gcgaagaaac aatcacaccg tggaacttcg aagaagtcgt cgacaaggga     1500

gcaagcgcac agagcttcat cgaaagaatg acaaacttcg acaagaacct gccgaacgaa     1560

aaggtcctgc cgaagcacag cctgctgtac gaatacttca cagtctacaa cgaactgaca     1620

aaggtcaagt acgtcacaga aggaatgaga aagccggcat tcctgagcgg agaacagaag     1680

aaggcaatcg tcgacctgct gttcaagaca aacagaaagg tcacagtcaa gcagctgaag     1740

gaagactact tcaagaagat cgaatgcttc gacagcgtcg aaatcagcgg agtcgaagac     1800

agattcaacg caagcctggg aacataccac gacctgctga agatcatcaa ggacaaggac     1860

ttcctggaca acgaagaaaa cgaagacatc ctggaagaca tcgtcctgac actgacactg     1920

ttcgaagaca gagaaatgat cgaagaaaga ctgaagacat acgcacacct gttcgacgac     1980

aaggtcatga agcagctgaa gagaagaaga tacacaggat ggggaagact gagcagaaag     2040

ctgatcaacg gaatcagaga caagcagagc ggaaagacaa tcctggactt cctgaagagc     2100

gacggattcg caaacagaaa cttcatgcag ctgatccacg acgacagcct gacattcaag     2160

gaagacatcc agaaggcaca ggtcagcgga cagggagaca gcctgcacga acacatcgca     2220

aacctggcag gaagcccggc aatcaagaag ggaatcctgc agacagtcaa ggtcgtcgac     2280

gaactggtca aggtcatggg aagacacaag ccggaaaaca tcgtcatcga aatggcaaga     2340

gaaaaccaga caacacagaa gggacagaag aacagcagag aaagaatgaa gagaatcgaa     2400

gaaggaatca aggaactggg aagccagatc ctgaaggaac acccggtcga aaacacacag     2460

ctgcagaacg aaaagctgta cctgtactac ctgcagaacg gaagagacat gtacgtcgac     2520

caggaactgg acatcaacag actgagcgac tacgacgtcg accacatcgt cccgcagagc     2580

ttcctgaagg acgacagcat cgacaacaag gtcctgacaa gaagcgacaa gaacagagga     2640

aagagcgaca acgtcccgag cgaagaagtc gtcaagaaga tgaagaacta ctggagacag     2700

ctgctgaacg caaagctgat cacacagaga aagttcgaca acctgacaaa ggcagagaga     2760

ggaggactga gcgaactgga caaggcagga ttcatcaaga gacagctggt cgaaacaaga     2820

cagatcacaa agcacgtcgc acagatcctg gacagcagaa tgaacacaaa gtacgacgaa     2880

aacgacaagc tgatcagaga agtcaaggtc atcacactga agagcaagct ggtcagcgac     2940

ttcagaaagg acttccagtt ctacaaggtc agagaaatca acaactacca ccacgcacac     3000

gacgcatacc tgaacgcagt cgtcggaaca gcactgatca agaagtaccc gaagctggaa     3060

agcgaattcg tctacggaga ctacaaggtc tacgacgtca gaaagatgat cgcaaagagc     3120

gaacaggaaa tcggaaaggc aacagcaaag tacttcttct acagcaacat catgaacttc     3180

ttcaagacag aaatcacact ggcaaacgga gaaatcagaa agagaccgct gatcgaaaca     3240

aacggagaaa caggagaaat cgtctgggac aagggaagag acttcgcaac agtcagaaag     3300

gtcctgagca tgccgcaggt caacatcgtc aagaagacag aagtccagac aggaggattc     3360

agcaaggaaa gcatcctgcc gaagagaaac agcgacaagc tgatcgcaag aaagaaggac     3420

tgggacccga agaagtacgg aggattcgac agcccgacag tcgcatacag cgtcctggtc     3480

gtcgcaaagg tcgaaaaggg aaagagcaag aagctgaaga gcgtcaagga actgctggga     3540

atcacaatca tggaaagaag cagcttcgaa aagaacccga tcgacttcct ggaagcaaag     3600

ggatacaagg aagtcaagaa ggacctgatc atcaagctgc cgaagtacag cctgttcgaa     3660

ctggaaaacg gaagaaagag aatgctggca agcgcaggag aactgcagaa gggaaacgaa     3720

ctggcactgc cgagcaagta cgtcaacttc ctgtacctgg caagccacta cgaaaagctg     3780

aagggaagcc cggaagacaa cgaacagaag cagctgttcg tcgaacagca caagcactac     3840

ctggacgaaa tcatcgaaca gatcagcgaa ttcagcaaga gagtcatcct ggcagacgca     3900

aacctggaca aggtcctgag cgcatacaac aagcacagag acaagccgat cagagaacag     3960

gcagaaaaca tcatccacct gttcacactg acaaacctgg gagcaccggc agcattcaag     4020

tacttcgaca caacaatcga cagaaagaga tacacaagca caaaggaagt cctggacgca     4080

acactgatcc accagagcat cacaggactg tacgaaacaa gaatcgacct gagccagctg     4140

ggaggagacg gaggaggaag cccgaagaag aagagaaagg tcagcgaaag cgcaacaccg     4200

gaaagcgtca gcggatggag actgttcaag aagatcagct agctagccat cacatttaaa     4260

agcatctcag cctaccatga gaataagaga aagaaaatga agatcaatag cttattcatc     4320

tctttttctt tttcgttggt gtaaagccaa caccctgtct aaaaaacata aatttcttta     4380

atcattttgc ctcttttctc tgtgcttcaa ttaataaaaa atggaaagaa cctcgagaaa     4440

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa     4500

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaa                              4537


<210>  186
<211>  1381
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS1

<400>  186

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Leu Ala Ala  Lys Arg Ser Arg Thr  Thr 
    1370                 1375                 1380     


<210>  187
<211>  1381
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS2

<400>  187

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Gln Ala Ala  Lys Arg Ser Arg Thr  Thr 
    1370                 1375                 1380     


<210>  188
<211>  1382
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS3

<400>  188

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Pro Ala Pro  Ala Lys Arg Glu Arg  Thr Thr 
    1370                 1375                 1380         


<210>  189
<211>  1381
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS4

<400>  189

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Gln Ala Ala  Lys Arg Pro Arg Thr  Thr 
    1370                 1375                 1380     


<210>  190
<211>  1381
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS5

<400>  190

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Arg Ala Ala  Lys Arg Pro Arg Thr  Thr 
    1370                 1375                 1380     


<210>  191
<211>  1383
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS6

<400>  191

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Ala Ala Ala  Lys Arg Ser Trp Ser  Met Ala Ala 
    1370                 1375                 1380             


<210>  192
<211>  1383
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS7

<400>  192

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Ala Ala Ala  Lys Arg Val Trp Ser  Met Ala Phe 
    1370                 1375                 1380             


<210>  193
<211>  1383
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS8

<400>  193

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Ala Ala Ala  Lys Arg Ser Trp Ser  Met Ala Phe 
    1370                 1375                 1380             


<210>  194
<211>  1382
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS9

<400>  194

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Ala Ala Ala  Lys Arg Lys Tyr Phe  Ala Ala 
    1370                 1375                 1380         


<210>  195
<211>  1382
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS10

<400>  195

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Arg Ala Ala  Lys Arg Lys Ala Phe  Ala Ala 
    1370                 1375                 1380         


<210>  196
<211>  1382
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Amino acid sequence for Cas9 with NLS11

<400>  196

Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val 
1               5                   10                  15      


Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe 
            20                  25                  30          


Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile 
        35                  40                  45              


Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu 
    50                  55                  60                  


Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys 
65                  70                  75                  80  


Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser 
                85                  90                  95      


Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys 
            100                 105                 110         


His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr 
        115                 120                 125             


His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp 
    130                 135                 140                 


Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His 
145                 150                 155                 160 


Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro 
                165                 170                 175     


Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr 
            180                 185                 190         


Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala 
        195                 200                 205             


Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn 
    210                 215                 220                 


Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn 
225                 230                 235                 240 


Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe 
                245                 250                 255     


Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp 
            260                 265                 270         


Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp 
        275                 280                 285             


Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp 
    290                 295                 300                 


Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser 
305                 310                 315                 320 


Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys 
                325                 330                 335     


Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe 
            340                 345                 350         


Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser 
        355                 360                 365             


Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp 
    370                 375                 380                 


Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg 
385                 390                 395                 400 


Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu 
                405                 410                 415     


Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe 
            420                 425                 430         


Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile 
        435                 440                 445             


Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp 
    450                 455                 460                 


Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu 
465                 470                 475                 480 


Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr 
                485                 490                 495     


Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser 
            500                 505                 510         


Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys 
        515                 520                 525             


Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln 
    530                 535                 540                 


Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr 
545                 550                 555                 560 


Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp 
                565                 570                 575     


Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly 
            580                 585                 590         


Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp 
        595                 600                 605             


Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr 
    610                 615                 620                 


Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala 
625                 630                 635                 640 


His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr 
                645                 650                 655     


Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp 
            660                 665                 670         


Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe 
        675                 680                 685             


Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe 
    690                 695                 700                 


Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu 
705                 710                 715                 720 


His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly 
                725                 730                 735     


Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly 
            740                 745                 750         


Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln 
        755                 760                 765             


Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile 
    770                 775                 780                 


Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro 
785                 790                 795                 800 


Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu 
                805                 810                 815     


Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg 
            820                 825                 830         


Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys 
        835                 840                 845             


Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg 
    850                 855                 860                 


Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys 
865                 870                 875                 880 


Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys 
                885                 890                 895     


Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp 
            900                 905                 910         


Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr 
        915                 920                 925             


Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp 
    930                 935                 940                 


Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser 
945                 950                 955                 960 


Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg 
                965                 970                 975     


Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val 
            980                 985                 990         


Val Gly Thr Ala Leu Ile Lys Lys  Tyr Pro Lys Leu Glu  Ser Glu Phe 
        995                 1000                 1005             


Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val Arg Lys  Met Ile Ala 
    1010                 1015                 1020             


Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr Ala Lys  Tyr Phe Phe 
    1025                 1030                 1035             


Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr Glu Ile  Thr Leu Ala 
    1040                 1045                 1050             


Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile Glu Thr  Asn Gly Glu 
    1055                 1060                 1065             


Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg Asp Phe  Ala Thr Val 
    1070                 1075                 1080             


Arg Lys  Val Leu Ser Met Pro  Gln Val Asn Ile Val  Lys Lys Thr 
    1085                 1090                 1095             


Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu Ser Ile  Leu Pro Lys 
    1100                 1105                 1110             


Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys Lys Asp  Trp Asp Pro 
    1115                 1120                 1125             


Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr Val Ala  Tyr Ser Val 
    1130                 1135                 1140             


Leu Val  Val Ala Lys Val Glu  Lys Gly Lys Ser Lys  Lys Leu Lys 
    1145                 1150                 1155             


Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile Met Glu  Arg Ser Ser 
    1160                 1165                 1170             


Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu Ala Lys  Gly Tyr Lys 
    1175                 1180                 1185             


Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu Pro Lys  Tyr Ser Leu 
    1190                 1195                 1200             


Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met Leu Ala  Ser Ala Gly 
    1205                 1210                 1215             


Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu Pro Ser  Lys Tyr Val 
    1220                 1225                 1230             


Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu Lys Leu  Lys Gly Ser 
    1235                 1240                 1245             


Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe Val Glu  Gln His Lys 
    1250                 1255                 1260             


His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile Ser Glu  Phe Ser Lys 
    1265                 1270                 1275             


Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp Lys Val  Leu Ser Ala 
    1280                 1285                 1290             


Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg Glu Gln  Ala Glu Asn 
    1295                 1300                 1305             


Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu Gly Ala  Pro Ala Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Arg  Tyr Thr Ser 
    1325                 1330                 1335             


Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile His Gln  Ser Ile Thr 
    1340                 1345                 1350             


Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser Gln Leu  Gly Gly Asp 
    1355                 1360                 1365             


Gly Gly  Gly Ser Arg Ala Ala  Lys Arg Lys Tyr Phe  Ala Val 
    1370                 1375                 1380         


<210>  197
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G506 guide RNA


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(97)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  197
auaccagucc agcgaggcag guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


<210>  198
<211>  100
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: G510 guide RNA


<220>
<221>  modified_base
<222>  (1)..(3)
<223>  PS linkage, 2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (29)..(40)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (69)..(96)
<223>  2'-O-Me nucleotide

<220>
<221>  modified_base
<222>  (97)..(100)
<223>  PS linkage, 2'-O-Me nucleotide

<400>  198
acuugucuuc ucuauaccca guuuuagagc uagaaauagc aaguuaaaau aaggcuaguc       60

cguuaucaac uugaaaaagu ggcaccgagu cggugcuuuu                            100


