                         SEQUENCE LISTING

<110>  Redbiotec AG
 
<120>  BACTERIAL DELIVERY OF GENE EDITING TOOLS INTO EUKARYOTIC CELLS

<130>  RBT16703PCT

<160>  2     

<170>  PatentIn version 3.5

<210>  1
<211>  1367
<212>  PRT
<213>  artificial

<220>
<223>  Engineered Cas9

<400>  1

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  2
<211>  4101
<212>  DNA
<213>  artificial

<220>
<223>  nucleotide sequence encoding engineered Cas9

<400>  2
gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc      180

acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat      240

ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg      300

gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg      660

ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg      720

attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat      780

gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg      900

ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg      960

atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag     1020

cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa     1140

aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc     1260

attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag     1320

aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga     1380

ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg     1440

gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat     1560

aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc     1620

ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg     1680

aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc     1740

ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc     1800

aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg     1860

accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac     1920

ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat     2040

ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac     2160

gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg     2220

aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg     2340

aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg     2400

gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat     2460

atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc     2520

gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac     2580

aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc     2700

aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg     2760

gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact     2820

aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag     2880

ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac     2940

caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac     3000

cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg     3060

atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac     3120

atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct     3180

ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc     3240

accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag     3300

acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc     3360

agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat     3420

tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa     3480

gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt     3540

ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac     3600

tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag     3660

aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac     3720

tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc     3840

ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc     3900

atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct     3960

gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac     4080

ctgtctcagc tgggaggcga c                                               4101


