                         SEQUENCE LISTING


<110> BASF Agricultural Solutions Seed US LLC

<120>  Codon-optimized Cas9 endonuclease encoding polynucleotide

<130>  191619WO01 

<160>  4

<170>  According Wipo Std 25






<210> 1
<211> 4107
<212> DNA
<213> Artificial sequence

<220>
<223> Codon optimized (wheat) cas9 gene

<400> 1
atggacaaga agtactccat cggcctcgac atcggcacca acagcgtcgg ctgggccgtg     60
atcaccgacg agtacaaggt gccaagcaag aagttcaagg tgctgggcaa caccgaccgt    120
cactccatca agaagaacct catcggcgcc ctcctcttcg actcaggtga gacggccgag    180
gcgacaaggt tgaagaggac ggcaagaaga aggtacacaa gaaggaagaa caggatctgc    240
tacctccagg agatcttcag caacgagatg gccaaggtgg atgacagctt cttccaccgg    300
ctagaggaga gcttcttggt ggaggaggac aagaagcatg agaggcatcc catcttcggc    360
aacatcgtcg acgaggtggc ctaccatgag aagtacccca ccatctacca tctccgcaag    420
aagctggtgg actcaacgga caaggccgac ctccgcctca tctacctcgc cttggcccac    480
atgatcaagt tcagaggcca cttcctcatc gagggcgacc tcaaccctga caacagcgat    540
gtggacaagc tcttcatcca gctggtgcag acctacaacc agctctttga ggagaacccc    600
atcaacgcct ccggcgtcga cgccaaggcc atcctctcgg caaggttgag caagtcaaga    660
aggctggaga acctcatcgc tcagctcccc ggcgagaaga aaaatggcct cttcggcaac    720
ctcatcgcct tgagcctcgg cctcaccccc aacttcaagt caaactttga cctcgccgag    780
gatgccaagc tccagctctc caaggacacc tacgacgacg acctcgacaa cctcctcgcc    840
cagatcggcg accagtacgc cgacctcttc ctcgccgcca agaacctctc agatgccatc    900
ctcctgagcg acatcctccg cgtcaacacc gagatcacca aggcgccgct ctccgcctcc    960
atgatcaaga ggtatgatga gcaccatcaa gatctcaccc tcctcaaggc cttggtgagg   1020
cagcagctac ctgagaagta caaggagatc ttcttcgacc agagcaagaa tggctacgcc   1080
ggctacatcg acggcggcgc ctcccaagaa gagttctaca agttcatcaa gcccatcttg   1140
gagaagatgg atggcaccga ggagctactg gtgaagctca acagagaaga tctcctcagg   1200
aagcagagga ccttcgacaa cggcagcatc cctcaccaga tccaccttgg tgagctccat   1260
gccatcttga gaaggcaaga agacttctac cccttcctca aggacaacag ggagaagatc   1320
gagaagatat tgacattcag gatcccctac tacgtcggac ccttggcaag aggcaactca   1380
aggttcgcct ggatgacaag gaagagcgag gaaaccatca ccccctggaa ctttgaggag   1440
gtggtggaca agggcgcctc agctcagagc ttcatcgaga ggatgaccaa cttcgacaag   1500
aacctcccca acgagaaggt gctccccaag cacagcctcc tctacgagta cttcaccgtc   1560
tacaatgagc tcaccaaggt caagtacgtc accgagggca tgaggaagcc ggccttcctc   1620
tcaggagagc agaagaaggc catcgtcgac ctcctcttca agaccaacag gaaggtcacc   1680
gtcaagcagc tcaaggagga ctacttcaag aagatcgagt gcttcgactc tgtggagatc   1740
tccggcgtgg aggacaggtt caacgcctcc ctcggcacct accatgatct cctgaagatc   1800
atcaaggaca aggacttcct cgacaacgag gagaacgagg acatcttaga agacatcgtc   1860
ctcaccctca ccctcttcga ggacagggag atgatcgagg agaggctcaa gacctatgct   1920
cacctctttg atgacaaggt gatgaagcag ctcaagagaa gaaggtacac cggctggggc   1980
cgcctctcaa ggaagctcat caacggcatc agggacaagc agagcggcaa gaccatcctc   2040
gacttcctca agagcgatgg cttcgccaac aggaacttca tgcagctcat ccatgatgac   2100
agcttgacct tcaaggagga catccagaag gctcaagtct ccggccaagg agattccctc   2160
catgagcaca tcgccaacct cgccggctca ccggccatca agaagggcat cctccagacc   2220
gtcaaggtgg tggatgagct ggtgaaggtg atgggccgtc acaagccgga gaacatcgtc   2280
atcgagatgg caagggagaa ccagaccacc cagaagggcc agaagaactc aagggagagg   2340
atgaagagga ttgaagaagg catcaaggag ctcggcagcc agatcctcaa ggagcaccct   2400
gtggagaaca cccagctaca gaacgagaag ctctacctct actacctcca gaatggccgt   2460
gacatgtacg tcgaccaaga gctggacatc aacaggttga gcgactacga cgtcgaccac   2520
atcgtccccc agagcttcct caaggatgac agcatcgaca acaaggtgct gacaagatca   2580
gacaagaacc gtggcaagag cgacaatgtc ccctccgagg aggtggtgaa gaagatgaag   2640
aactactgga ggcagctcct caacgccaag ctcatcaccc agaggaagtt cgacaacctc   2700
accaaggctg aaagaggagg cctctcagag ctggacaagg ccggcttcat caagaggcag   2760
ctggtggaga caaggcagat caccaagcat gttgctcaga tccttgattc aaggatgaac   2820
accaagtacg acgagaacga caagctcatc agggaggtca aggtcatcac cctcaagagc   2880
aagctggtgt ccgacttcag gaaggacttc cagttctaca aggtgaggga gatcaacaac   2940
taccaccatg ctcatgatgc ctacctcaac gccgtggtgg gcaccgccct catcaagaag   3000
taccccaagc tggagtcaga gttcgtctac ggcgactaca aggtgtatga tgtgaggaag   3060
atgatcgcca agagcgagca ggagatcggc aaggccaccg ccaagtactt cttctactca   3120
aacatcatga acttcttcaa gaccgagatc accctcgcca acggcgagat caggaagagg   3180
ccgctcattg agacaaatgg tgagactggt gagatcgtct gggacaaggg ccgagacttc   3240
gccaccgtga ggaaggtgct ctccatgcct caagtcaaca tcgtcaagaa gaccgaggtg   3300
cagaccggag gcttctcaaa ggagagcatc ctccccaaga ggaactctga caagctgatc   3360
gcaaggaaga aggactggga ccccaagaag tacggcggct ttgattcacc aacggtggcc   3420
tactccgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct caagagcgtc   3480
aaggagctcc tcggcatcac catcatggag agatcatcct tcgagaagaa ccccatcgac   3540
ttccttgagg ccaagggcta caaggaggtg aagaaggacc tcatcatcaa gctccccaag   3600
tacagcctct tcgagctgga gaatggccgc aagaggatgc tggcatcagc tggagagctc   3660
cagaagggca acgagctggc cctcccaagc aagtacgtga acttcctcta cctcgcctcc   3720
cactacgaga agctcaaggg ctcaccagaa gacaatgagc agaagcagct cttcgtggag   3780
cagcacaagc actacctcga cgagatcatc gagcagatct ccgagttctc caagagggtg   3840
atcctcgccg acgccaacct cgacaaggtg ctctcagcct acaacaagca ccgtgacaag   3900
cccatcaggg agcaagctga gaacatcatc cacctcttca ccctcaccaa cctcggcgcc   3960
ccggcggcct tcaagtactt cgacaccacc atcgaccgca agaggtacac ctcaaccaag   4020
gaggtgcttg atgccacctt gatccaccag agcatcaccg gcctctacga gacaaggatc   4080
gacctctccc agctcggcgg cgactga                                       4107

<210> 2
<211> 1368
<212> PRT
<213> Artificial sequence

<220>
<223> Cas9 protein encoded by Seq-ID 1

<400> 2
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 Lys
    1010                1015                1020
Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser
1025                1030                1035                1040
Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu
                1045                1050                1055
Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile
            1060                1065                1070
Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser
        1075                1080                1085
Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly
    1090                1095                1100
Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile
1105                1110                1115                1120
Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser
                1125                1130                1135
Pro Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly
            1140                1145                1150
Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile
        1155                1160                1165
Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala
    1170                1175                1180
Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys
1185                1190                1195                1200
Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser
                1205                1210                1215
Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr
            1220                1225                1230
Val 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 His
    1250                1255                1260
Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val
1265                1270                1275                1280
Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys
                1285                1290                1295
His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His Leu
            1300                1305                1310
Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp
        1315                1320                1325
Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp
    1330                1335                1340
Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile
1345                1350                1355                1360
Asp Leu Ser Gln Leu Gly Gly Asp
                1365

<210> 3
<211> 4206
<212> DNA
<213> Artificial sequence

<220>
<223> Codon optimized (wheat) cas9 gene (with fused NLS)

<400> 3
atggcaccca agaagaagag gaaggtgggc atccacggcg tcccagcagc aatggacaag     60
aagtactcca tcggcctcga catcggcacc aacagcgtcg gctgggccgt gatcaccgac    120
gagtacaagg tgccaagcaa gaagttcaag gtgctgggca acaccgaccg tcactccatc    180
aagaagaacc tcatcggcgc cctcctcttc gactcaggtg agacggccga ggcgacaagg    240
ttgaagagga cggcaagaag aaggtacaca agaaggaaga acaggatctg ctacctccag    300
gagatcttca gcaacgagat ggccaaggtg gatgacagct tcttccaccg gctagaggag    360
agcttcttgg tggaggagga caagaagcat gagaggcatc ccatcttcgg caacatcgtc    420
gacgaggtgg cctaccatga gaagtacccc accatctacc atctccgcaa gaagctggtg    480
gactcaacgg acaaggccga cctccgcctc atctacctcg ccttggccca catgatcaag    540
ttcagaggcc acttcctcat cgagggcgac ctcaaccctg acaacagcga tgtggacaag    600
ctcttcatcc agctggtgca gacctacaac cagctctttg aggagaaccc catcaacgcc    660
tccggcgtcg acgccaaggc catcctctcg gcaaggttga gcaagtcaag aaggctggag    720
aacctcatcg ctcagctccc cggcgagaag aaaaatggcc tcttcggcaa cctcatcgcc    780
ttgagcctcg gcctcacccc caacttcaag tcaaactttg acctcgccga ggatgccaag    840
ctccagctct ccaaggacac ctacgacgac gacctcgaca acctcctcgc ccagatcggc    900
gaccagtacg ccgacctctt cctcgccgcc aagaacctct cagatgccat cctcctgagc    960
gacatcctcc gcgtcaacac cgagatcacc aaggcgccgc tctccgcctc catgatcaag   1020
aggtatgatg agcaccatca agatctcacc ctcctcaagg ccttggtgag gcagcagcta   1080
cctgagaagt acaaggagat cttcttcgac cagagcaaga atggctacgc cggctacatc   1140
gacggcggcg cctcccaaga agagttctac aagttcatca agcccatctt ggagaagatg   1200
gatggcaccg aggagctact ggtgaagctc aacagagaag atctcctcag gaagcagagg   1260
accttcgaca acggcagcat ccctcaccag atccaccttg gtgagctcca tgccatcttg   1320
agaaggcaag aagacttcta ccccttcctc aaggacaaca gggagaagat cgagaagata   1380
ttgacattca ggatccccta ctacgtcgga cccttggcaa gaggcaactc aaggttcgcc   1440
tggatgacaa ggaagagcga ggaaaccatc accccctgga actttgagga ggtggtggac   1500
aagggcgcct cagctcagag cttcatcgag aggatgacca acttcgacaa gaacctcccc   1560
aacgagaagg tgctccccaa gcacagcctc ctctacgagt acttcaccgt ctacaatgag   1620
ctcaccaagg tcaagtacgt caccgagggc atgaggaagc cggccttcct ctcaggagag   1680
cagaagaagg ccatcgtcga cctcctcttc aagaccaaca ggaaggtcac cgtcaagcag   1740
ctcaaggagg actacttcaa gaagatcgag tgcttcgact ctgtggagat ctccggcgtg   1800
gaggacaggt tcaacgcctc cctcggcacc taccatgatc tcctgaagat catcaaggac   1860
aaggacttcc tcgacaacga ggagaacgag gacatcttag aagacatcgt cctcaccctc   1920
accctcttcg aggacaggga gatgatcgag gagaggctca agacctatgc tcacctcttt   1980
gatgacaagg tgatgaagca gctcaagaga agaaggtaca ccggctgggg ccgcctctca   2040
aggaagctca tcaacggcat cagggacaag cagagcggca agaccatcct cgacttcctc   2100
aagagcgatg gcttcgccaa caggaacttc atgcagctca tccatgatga cagcttgacc   2160
ttcaaggagg acatccagaa ggctcaagtc tccggccaag gagattccct ccatgagcac   2220
atcgccaacc tcgccggctc accggccatc aagaagggca tcctccagac cgtcaaggtg   2280
gtggatgagc tggtgaaggt gatgggccgt cacaagccgg agaacatcgt catcgagatg   2340
gcaagggaga accagaccac ccagaagggc cagaagaact caagggagag gatgaagagg   2400
attgaagaag gcatcaagga gctcggcagc cagatcctca aggagcaccc tgtggagaac   2460
acccagctac agaacgagaa gctctacctc tactacctcc agaatggccg tgacatgtac   2520
gtcgaccaag agctggacat caacaggttg agcgactacg acgtcgacca catcgtcccc   2580
cagagcttcc tcaaggatga cagcatcgac aacaaggtgc tgacaagatc agacaagaac   2640
cgtggcaaga gcgacaatgt cccctccgag gaggtggtga agaagatgaa gaactactgg   2700
aggcagctcc tcaacgccaa gctcatcacc cagaggaagt tcgacaacct caccaaggct   2760
gaaagaggag gcctctcaga gctggacaag gccggcttca tcaagaggca gctggtggag   2820
acaaggcaga tcaccaagca tgttgctcag atccttgatt caaggatgaa caccaagtac   2880
gacgagaacg acaagctcat cagggaggtc aaggtcatca ccctcaagag caagctggtg   2940
tccgacttca ggaaggactt ccagttctac aaggtgaggg agatcaacaa ctaccaccat   3000
gctcatgatg cctacctcaa cgccgtggtg ggcaccgccc tcatcaagaa gtaccccaag   3060
ctggagtcag agttcgtcta cggcgactac aaggtgtatg atgtgaggaa gatgatcgcc   3120
aagagcgagc aggagatcgg caaggccacc gccaagtact tcttctactc aaacatcatg   3180
aacttcttca agaccgagat caccctcgcc aacggcgaga tcaggaagag gccgctcatt   3240
gagacaaatg gtgagactgg tgagatcgtc tgggacaagg gccgagactt cgccaccgtg   3300
aggaaggtgc tctccatgcc tcaagtcaac atcgtcaaga agaccgaggt gcagaccgga   3360
ggcttctcaa aggagagcat cctccccaag aggaactctg acaagctgat cgcaaggaag   3420
aaggactggg accccaagaa gtacggcggc tttgattcac caacggtggc ctactccgtg   3480
ctggtggtgg ccaaggtgga gaagggcaag agcaagaagc tcaagagcgt caaggagctc   3540
ctcggcatca ccatcatgga gagatcatcc ttcgagaaga accccatcga cttccttgag   3600
gccaagggct acaaggaggt gaagaaggac ctcatcatca agctccccaa gtacagcctc   3660
ttcgagctgg agaatggccg caagaggatg ctggcatcag ctggagagct ccagaagggc   3720
aacgagctgg ccctcccaag caagtacgtg aacttcctct acctcgcctc ccactacgag   3780
aagctcaagg gctcaccaga agacaatgag cagaagcagc tcttcgtgga gcagcacaag   3840
cactacctcg acgagatcat cgagcagatc tccgagttct ccaagagggt gatcctcgcc   3900
gacgccaacc tcgacaaggt gctctcagcc tacaacaagc accgtgacaa gcccatcagg   3960
gagcaagctg agaacatcat ccacctcttc accctcacca acctcggcgc cccggcggcc   4020
ttcaagtact tcgacaccac catcgaccgc aagaggtaca cctcaaccaa ggaggtgctt   4080
gatgccacct tgatccacca gagcatcacc ggcctctacg agacaaggat cgacctctcc   4140
cagctcggcg gcgacaagag gccggcggcc accaagaagg ccggccaagc caagaagaag   4200
aagtga                                                              4206

<210> 4
<211> 1401
<212> PRT
<213> Artificial sequence

<220>
<223> Cas9 protein (with fused NLS) encoded by Seq-ID 3

<400> 4
Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His Gly Val Pro Ala
1               5                   10                  15
Ala Met Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser
            20                  25                  30
Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys
        35                  40                  45
Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu
    50                  55                  60
Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg
65                  70                  75                  80
Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile
                85                  90                  95
Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp
            100                 105                 110
Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys
        115                 120                 125
Lys His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala
    130                 135                 140
Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val
145                 150                 155                 160
Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala
                165                 170                 175
His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn
            180                 185                 190
Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr
        195                 200                 205
Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp
    210                 215                 220
Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu
225                 230                 235                 240
Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly
                245                 250                 255
Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn
            260                 265                 270
Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr
        275                 280                 285
Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala
    290                 295                 300
Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser
305                 310                 315                 320
Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala
                325                 330                 335
Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu
            340                 345                 350
Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe
        355                 360                 365
Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala
    370                 375                 380
Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met
385                 390                 395                 400
Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu
                405                 410                 415
Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His
            420                 425                 430
Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro
        435                 440                 445
Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg
    450                 455                 460
Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala
465                 470                 475                 480
Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu
                485                 490                 495
Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met
            500                 505                 510
Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His
        515                 520                 525
Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val
    530                 535                 540
Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu
545                 550                 555                 560
Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val
                565                 570                 575
Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe
            580                 585                 590
Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu
        595                 600                 605
Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu
    610                 615                 620
Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu
625                 630                 635                 640
Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr
                645                 650                 655
Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg
            660                 665                 670
Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg
        675                 680                 685
Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly
    690                 695                 700
Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr
705                 710                 715                 720
Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser
                725                 730                 735
Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys
            740                 745                 750
Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met
        755                 760                 765
Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn
    770                 775                 780
Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg
785                 790                 795                 800
Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His
                805                 810                 815
Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr
            820                 825                 830
Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn
        835                 840                 845
Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu
    850                 855                 860
Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn
865                 870                 875                 880
Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met
                885                 890                 895
Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg
            900                 905                 910
Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu
        915                 920                 925
Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile
    930                 935                 940
Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr
945                 950                 955                 960
Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys
                965                 970                 975
Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val
            980                 985                 990
Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala
        995                 1000                1005
Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu
    1010                1015                1020
Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala
1025                1030                1035                1040
Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr
                1045                1050                1055
Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly
            1060                1065                1070
Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu
        1075                1080                1085
Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu
    1090                1095                1100
Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly
1105                1110                1115                1120
Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu
                1125                1130                1135
Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp
            1140                1145                1150
Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys
        1155                1160                1165
Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr
    1170                1175                1180
Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu
1185                1190                1195                1200
Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro
                1205                1210                1215
Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala
            1220                1225                1230
Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys
        1235                1240                1245
Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly
    1250                1255                1260
Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys
1265                1270                1275                1280
His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg
                1285                1290                1295
Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn
            1300                1305                1310
Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His
        1315                1320                1325
Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe
    1330                1335                1340
Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu
1345                1350                1355                1360
Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg
                1365                1370                1375
Ile Asp Leu Ser Gln Leu Gly Gly Asp Lys Arg Pro Ala Ala Thr Lys
            1380                1385                1390
Lys Ala Gly Gln Ala Lys Lys Lys Lys
        1395                1400
