                         SEQUENCE LISTING

<110>  University of Copenhagen
 
<120>  CRISPR/Cas screening system, materials and methods

<130>  P5743PC00

<160>  10    

<170>  PatentIn version 3.5

<210>  1
<211>  1368
<212>  PRT
<213>  Streptococcus pyogenes

<400>  1

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>  2
<211>  1399
<212>  PRT
<213>  Artificial

<220>
<223>  Human, codon-optimized (D10A) nickase


<220>
<221>  MISC_FEATURE
<222>  (1)..(1399)
<223>  Sequence is in silico translation of Cas9-D10A in the plasmid 
       pCas9n-sgPdx1

<400>  2

Met Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Pro Lys Lys Lys Arg 
1               5                   10                  15      


Lys Val Glu Ala Ser Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly 
            20                  25                  30          


Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro 
        35                  40                  45              


Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys 
    50                  55                  60                  


Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu 
65                  70                  75                  80  


Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys 
                85                  90                  95      


Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys 
            100                 105                 110         


Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu 
        115                 120                 125             


Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp 
    130                 135                 140                 


Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys 
145                 150                 155                 160 


Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu 
                165                 170                 175     


Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly 
            180                 185                 190         


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


Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser 
    210                 215                 220                 


Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg 
225                 230                 235                 240 


Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly 
                245                 250                 255     


Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe 
            260                 265                 270         


Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys 
        275                 280                 285             


Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp 
    290                 295                 300                 


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


Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro 
                325                 330                 335     


Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu 
            340                 345                 350         


Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys 
        355                 360                 365             


Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp 
    370                 375                 380                 


Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu 
385                 390                 395                 400 


Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu 
                405                 410                 415     


Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His 
            420                 425                 430         


Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp 
        435                 440                 445             


Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu 
    450                 455                 460                 


Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser 
465                 470                 475                 480 


Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp 
                485                 490                 495     


Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile 
            500                 505                 510         


Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu 
        515                 520                 525             


Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu 
    530                 535                 540                 


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


Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn 
                565                 570                 575     


Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile 
            580                 585                 590         


Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn 
        595                 600                 605             


Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys 
    610                 615                 620                 


Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val 
625                 630                 635                 640 


Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu 
                645                 650                 655     


Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys 
            660                 665                 670         


Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn 
        675                 680                 685             


Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys 
    690                 695                 700                 


Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp 
705                 710                 715                 720 


Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln 
                725                 730                 735     


Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala 
            740                 745                 750         


Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val 
        755                 760                 765             


Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala 
    770                 775                 780                 


Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg 
785                 790                 795                 800 


Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu 
                805                 810                 815     


Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr 
            820                 825                 830         


Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu 
        835                 840                 845             


Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln 
    850                 855                 860                 


Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser 
865                 870                 875                 880 


Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val 
                885                 890                 895     


Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile 
            900                 905                 910         


Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu 
        915                 920                 925             


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


Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn 
945                 950                 955                 960 


Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile 
                965                 970                 975     


Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe 
            980                 985                 990         


Tyr Lys Val Arg Glu Ile Asn Asn  Tyr His His Ala His  Asp Ala Tyr 
        995                 1000                 1005             


Leu Asn  Ala Val Val Gly Thr  Ala Leu Ile Lys Lys  Tyr Pro Lys 
    1010                 1015                 1020             


Leu Glu  Ser Glu Phe Val Tyr  Gly Asp Tyr Lys Val  Tyr Asp Val 
    1025                 1030                 1035             


Arg Lys  Met Ile Ala Lys Ser  Glu Gln Glu Ile Gly  Lys Ala Thr 
    1040                 1045                 1050             


Ala Lys  Tyr Phe Phe Tyr Ser  Asn Ile Met Asn Phe  Phe Lys Thr 
    1055                 1060                 1065             


Glu Ile  Thr Leu Ala Asn Gly  Glu Ile Arg Lys Arg  Pro Leu Ile 
    1070                 1075                 1080             


Glu Thr  Asn Gly Glu Thr Gly  Glu Ile Val Trp Asp  Lys Gly Arg 
    1085                 1090                 1095             


Asp Phe  Ala Thr Val Arg Lys  Val Leu Ser Met Pro  Gln Val Asn 
    1100                 1105                 1110             


Ile Val  Lys Lys Thr Glu Val  Gln Thr Gly Gly Phe  Ser Lys Glu 
    1115                 1120                 1125             


Ser Ile  Leu Pro Lys Arg Asn  Ser Asp Lys Leu Ile  Ala Arg Lys 
    1130                 1135                 1140             


Lys Asp  Trp Asp Pro Lys Lys  Tyr Gly Gly Phe Asp  Ser Pro Thr 
    1145                 1150                 1155             


Val Ala  Tyr Ser Val Leu Val  Val Ala Lys Val Glu  Lys Gly Lys 
    1160                 1165                 1170             


Ser Lys  Lys Leu Lys Ser Val  Lys Glu Leu Leu Gly  Ile Thr Ile 
    1175                 1180                 1185             


Met Glu  Arg Ser Ser Phe Glu  Lys Asn Pro Ile Asp  Phe Leu Glu 
    1190                 1195                 1200             


Ala Lys  Gly Tyr Lys Glu Val  Lys Lys Asp Leu Ile  Ile Lys Leu 
    1205                 1210                 1215             


Pro Lys  Tyr Ser Leu Phe Glu  Leu Glu Asn Gly Arg  Lys Arg Met 
    1220                 1225                 1230             


Leu Ala  Ser Ala Gly Glu Leu  Gln Lys Gly Asn Glu  Leu Ala Leu 
    1235                 1240                 1245             


Pro Ser  Lys Tyr Val Asn Phe  Leu Tyr Leu Ala Ser  His Tyr Glu 
    1250                 1255                 1260             


Lys Leu  Lys Gly Ser Pro Glu  Asp Asn Glu Gln Lys  Gln Leu Phe 
    1265                 1270                 1275             


Val Glu  Gln His Lys His Tyr  Leu Asp Glu Ile Ile  Glu Gln Ile 
    1280                 1285                 1290             


Ser Glu  Phe Ser Lys Arg Val  Ile Leu Ala Asp Ala  Asn Leu Asp 
    1295                 1300                 1305             


Lys Val  Leu Ser Ala Tyr Asn  Lys His Arg Asp Lys  Pro Ile Arg 
    1310                 1315                 1320             


Glu Gln  Ala Glu Asn Ile Ile  His Leu Phe Thr Leu  Thr Asn Leu 
    1325                 1330                 1335             


Gly Ala  Pro Ala Ala Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg 
    1340                 1345                 1350             


Lys Arg  Tyr Thr Ser Thr Lys  Glu Val Leu Asp Ala  Thr Leu Ile 
    1355                 1360                 1365             


His Gln  Ser Ile Thr Gly Leu  Tyr Glu Thr Arg Ile  Asp Leu Ser 
    1370                 1375                 1380             


Gln Leu  Gly Gly Asp Ser Pro  Lys Lys Lys Arg Lys  Val Glu Ala 
    1385                 1390                 1395             


Ser 
    


<210>  3
<211>  1300
<212>  PRT
<213>  Francisella novicida

<400>  3

Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr 
1               5                   10                  15      


Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys 
            20                  25                  30          


Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys 
        35                  40                  45              


Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu 
    50                  55                  60                  


Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser 
65                  70                  75                  80  


Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys 
                85                  90                  95      


Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr 
            100                 105                 110         


Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile 
        115                 120                 125             


Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln 
    130                 135                 140                 


Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr 
145                 150                 155                 160 


Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr 
                165                 170                 175     


Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser 
            180                 185                 190         


Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu 
        195                 200                 205             


Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys 
    210                 215                 220                 


Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu 
225                 230                 235                 240 


Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg 
                245                 250                 255     


Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr 
            260                 265                 270         


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


Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile 
    290                 295                 300                 


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


Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser 
                325                 330                 335     


Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met 
            340                 345                 350         


Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys 
        355                 360                 365             


Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln 
    370                 375                 380                 


Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr 
385                 390                 395                 400 


Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala 
                405                 410                 415     


Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn 
            420                 425                 430         


Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala 
        435                 440                 445             


Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn 
    450                 455                 460                 


Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala 
465                 470                 475                 480 


Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys 
                485                 490                 495     


Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys 
            500                 505                 510         


Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp 
        515                 520                 525             


Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His 
    530                 535                 540                 


Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His 
545                 550                 555                 560 


Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val 
                565                 570                 575     


Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser 
            580                 585                 590         


Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly 
        595                 600                 605             


Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys 
    610                 615                 620                 


Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile 
625                 630                 635                 640 


Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys 
                645                 650                 655     


Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val 
            660                 665                 670         


Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile 
        675                 680                 685             


Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln 
    690                 695                 700                 


Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe 
705                 710                 715                 720 


Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp 
                725                 730                 735     


Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu 
            740                 745                 750         


Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn 
        755                 760                 765             


Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr 
    770                 775                 780                 


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


Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn 
                805                 810                 815     


Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr 
            820                 825                 830         


Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala 
        835                 840                 845             


Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu 
    850                 855                 860                 


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


His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe 
                885                 890                 895     


Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His 
            900                 905                 910         


Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu 
        915                 920                 925             


Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile 
    930                 935                 940                 


Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile 
945                 950                 955                 960 


Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn 
                965                 970                 975     


Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile 
            980                 985                 990         


Ala Lys Leu Val Ile Glu Tyr Asn  Ala Ile Val Val Phe  Glu Asp Leu 
        995                 1000                 1005             


Asn Phe  Gly Phe Lys Arg Gly  Arg Phe Lys Val Glu  Lys Gln Val 
    1010                 1015                 1020             


Tyr Gln  Lys Leu Glu Lys Met  Leu Ile Glu Lys Leu  Asn Tyr Leu 
    1025                 1030                 1035             


Val Phe  Lys Asp Asn Glu Phe  Asp Lys Thr Gly Gly  Val Leu Arg 
    1040                 1045                 1050             


Ala Tyr  Gln Leu Thr Ala Pro  Phe Glu Thr Phe Lys  Lys Met Gly 
    1055                 1060                 1065             


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


Lys Ile  Cys Pro Val Thr Gly  Phe Val Asn Gln Leu  Tyr Pro Lys 
    1085                 1090                 1095             


Tyr Glu  Ser Val Ser Lys Ser  Gln Glu Phe Phe Ser  Lys Phe Asp 
    1100                 1105                 1110             


Lys Ile  Cys Tyr Asn Leu Asp  Lys Gly Tyr Phe Glu  Phe Ser Phe 
    1115                 1120                 1125             


Asp Tyr  Lys Asn Phe Gly Asp  Lys Ala Ala Lys Gly  Lys Trp Thr 
    1130                 1135                 1140             


Ile Ala  Ser Phe Gly Ser Arg  Leu Ile Asn Phe Arg  Asn Ser Asp 
    1145                 1150                 1155             


Lys Asn  His Asn Trp Asp Thr  Arg Glu Val Tyr Pro  Thr Lys Glu 
    1160                 1165                 1170             


Leu Glu  Lys Leu Leu Lys Asp  Tyr Ser Ile Glu Tyr  Gly His Gly 
    1175                 1180                 1185             


Glu Cys  Ile Lys Ala Ala Ile  Cys Gly Glu Ser Asp  Lys Lys Phe 
    1190                 1195                 1200             


Phe Ala  Lys Leu Thr Ser Val  Leu Asn Thr Ile Leu  Gln Met Arg 
    1205                 1210                 1215             


Asn Ser  Lys Thr Gly Thr Glu  Leu Asp Tyr Leu Ile  Ser Pro Val 
    1220                 1225                 1230             


Ala Asp  Val Asn Gly Asn Phe  Phe Asp Ser Arg Gln  Ala Pro Lys 
    1235                 1240                 1245             


Asn Met  Pro Gln Asp Ala Asp  Ala Asn Gly Ala Tyr  His Ile Gly 
    1250                 1255                 1260             


Leu Lys  Gly Leu Met Leu Leu  Gly Arg Ile Lys Asn  Asn Gln Glu 
    1265                 1270                 1275             


Gly Lys  Lys Leu Asn Leu Val  Ile Lys Asn Glu Glu  Tyr Phe Glu 
    1280                 1285                 1290             


Phe Val  Gln Asn Arg Asn Asn  
    1295                 1300 


<210>  4
<211>  4104
<212>  DNA
<213>  Artificial

<220>
<223>  Cas9-NG


<220>
<221>  misc_feature
<222>  (1)..(4104)
<223>  Sequence derived from Cas9-NG in the plasmid pX330-SpCas9-NG

<400>  4
atggacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag      180

gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc      240

tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga      300

ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag      420

aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga      660

cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac      720

ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc      900

ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct      960

atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg     1020

cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg     1140

gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac     1260

gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc     1320

gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc     1380

agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa     1440

gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg     1620

agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc     1680

gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc     1740

tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt     1800

atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg     1860

ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc     1920

cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc     1980

aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg     2040

gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg     2160

cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca     2220

gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg     2280

atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga     2340

atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc     2400

gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg     2460

gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat     2520

atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc     2580

gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg     2700

accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag     2760

ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac     2820

actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc     2880

aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag     3000

taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc     3120

aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg     3180

cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt     3240

gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg     3300

cagacaggcg gcttcagcaa agagtctatc cggcccaaga ggaacagcga taagctgatc     3360

gccagaaaga aggactggga ccctaagaag tacggcggct tcgttagccc caccgtggcc     3420

tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg     3480

aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac     3540

tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag     3600

tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc ccggttcctg     3660

cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc     3720

cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag     3900

cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc     3960

cctcgggcct tcaagtactt tgacaccacc atcgaccgga aggtgtaccg gagcaccaaa     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgac                                            4104


<210>  5
<211>  1368
<212>  PRT
<213>  Artificial

<220>
<223>  Cas9-NG


<220>
<221>  MISC_FEATURE
<222>  (1)..(1368)
<223>  In silico translation based on DNA sequence of Cas9-NG in the 
       plasmid pX330-SpCas9-NG

<400>  5

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  Arg 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 Val  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 Arg 
    1205                 1210                 1215             


Phe 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 Arg Ala 
    1310                 1315                 1320             


Phe Lys  Tyr Phe Asp Thr Thr  Ile Asp Arg Lys Val  Tyr Arg 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>  6
<211>  4104
<212>  DNA
<213>  Streptococcus pyogenes

<400>  6
atggacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag      180

gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc      240

tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga      300

ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag      420

aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga      660

cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac      720

ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc      900

ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct      960

atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg     1020

cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg     1140

gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac     1260

gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc     1320

gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc     1380

agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa     1440

gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg     1620

agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc     1680

gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc     1740

tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt     1800

atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg     1860

ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc     1920

cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc     1980

aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg     2040

gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg     2160

cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca     2220

gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg     2280

atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga     2340

atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc     2400

gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg     2460

gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat     2520

atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc     2580

gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg     2700

accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag     2760

ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac     2820

actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc     2880

aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag     3000

taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc     3120

aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg     3180

cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt     3240

gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg     3300

cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc     3360

gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc     3420

tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg     3480

aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac     3540

tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag     3600

tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg     3660

cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc     3720

cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag     3900

cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc     3960

cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgac                                            4104


<210>  7
<211>  4104
<212>  DNA
<213>  Artificial

<220>
<223>  Human, codon-optimized (D10A) nickase


<220>
<221>  misc_feature
<222>  (1)..(4104)
<223>  Sequence is Cas9-D10A in the plasmid pCas9n-sgPdx1

<400>  7
atggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag      180

gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc      240

tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga      300

ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag      420

aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga      660

cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac      720

ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc      900

ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct      960

atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg     1020

cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg     1140

gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac     1260

gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc     1320

gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc     1380

agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa     1440

gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg     1620

agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc     1680

gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc     1740

tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt     1800

atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg     1860

ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc     1920

cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc     1980

aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg     2040

gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg     2160

cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca     2220

gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg     2280

atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga     2340

atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc     2400

gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg     2460

gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat     2520

atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc     2580

gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg     2700

accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag     2760

ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac     2820

actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc     2880

aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag     3000

taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc     3120

aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg     3180

cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt     3240

gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg     3300

cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc     3360

gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc     3420

tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg     3480

aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac     3540

tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag     3600

tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg     3660

cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc     3720

cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag     3900

cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc     3960

cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgac                                            4104


<210>  8
<211>  3960
<212>  DNA
<213>  Francisella novicida

<400>  8
atgtcaattt atcaagaatt tgttaataaa tatagtttaa gtaaaactct aagatttgag       60

ttaatcccac agggtaaaac acttgaaaac ataaaagcaa gaggtttgat tttagatgat      120

gagaaaagag ctaaagacta caaaaaggct aaacaaataa ttgataaata tcatcagttt      180

tttatagagg agatattaag ttcggtttgt attagcgaag atttattaca aaactattct      240

gatgtttatt ttaaacttaa aaagagtgat gatgataatc tacaaaaaga ttttaaaagt      300

gcaaaagata cgataaagaa acaaatatct gaatatataa aggactcaga gaaatttaag      360

aatttgttta atcaaaacct tatcgatgct aaaaaagggc aagagtcaga tttaattcta      420

tggctaaagc aatctaagga taatggtata gaactattta aagccaatag tgatatcaca      480

gatatagatg aggcgttaga aataatcaaa tcttttaaag gttggacaac ttattttaag      540

ggttttcatg aaaatagaaa aaatgtttat agtagcaatg atattcctac atctattatt      600

tataggatag tagatgataa tttgcctaaa tttctagaaa ataaagctaa gtatgagagt      660

ttaaaagaca aagctccaga agctataaac tatgaacaaa ttaaaaaaga tttggcagaa      720

gagctaacct ttgatattga ctacaaaaca tctgaagtta atcaaagagt tttttcactt      780

gatgaagttt ttgagatagc aaactttaat aattatctaa atcaaagtgg tattactaaa      840

tttaatacta ttattggtgg taaatttgta aatggtgaaa atacaaagag aaaaggtata      900

aatgaatata taaatctata ctcacagcaa ataaatgata aaacactcaa aaaatataaa      960

atgagtgttt tatttaagca aattttaagt gatacagaat ctaaatcttt tgtaattgat     1020

aagttagaag atgatagtga tgtagttaca acgatgcaaa gtttttatga gcaaatagca     1080

gcttttaaaa cagtagaaga aaaatctatt aaagaaacac tatctttatt atttgatgat     1140

ttaaaagctc aaaaacttga tttgagtaaa atttatttta aaaatgataa atctcttact     1200

gatctatcac aacaagtttt tgatgattat agtgttattg gtacagcggt actagaatat     1260

ataactcaac aaatagcacc taaaaatctt gataacccta gtaagaaaga gcaagaatta     1320

atagccaaaa aaactgaaaa agcaaaatac ttatctctag aaactataaa gcttgcctta     1380

gaagaattta ataagcatag agatatagat aaacagtgta ggtttgaaga aatacttgca     1440

aactttgcgg ctattccgat gatatttgat gaaatagctc aaaacaaaga caatttggca     1500

cagatatcta tcaaatatca aaatcaaggt aaaaaagacc tacttcaagc tagtgcggaa     1560

gatgatgtta aagctatcaa ggatctttta gatcaaacta ataatctctt acataaacta     1620

aaaatatttc atattagtca gtcagaagat aaggcaaata ttttagacaa ggatgagcat     1680

ttttatctag tatttgagga gtgctacttt gagctagcga atatagtgcc tctttataac     1740

aaaattagaa actatataac tcaaaagcca tatagtgatg agaaatttaa gctcaatttt     1800

gagaactcga ctttggctaa tggttgggat aaaaataaag agcctgacaa tacggcaatt     1860

ttatttatca aagatgataa atattatctg ggtgtgatga ataagaaaaa taacaaaata     1920

tttgatgata aagctatcaa agaaaataaa ggcgagggtt ataaaaaaat tgtttataaa     1980

cttttacctg gcgcaaataa aatgttacct aaggttttct tttctgctaa atctataaaa     2040

ttttataatc ctagtgaaga tatacttaga ataagaaatc attccacaca tacaaaaaat     2100

ggtagtcctc aaaaaggata tgaaaaattt gagtttaata ttgaagattg ccgaaaattt     2160

atagattttt ataaacagtc tataagtaag catccggagt ggaaagattt tggatttaga     2220

ttttctgata ctcaaagata taattctata gatgaatttt atagagaagt tgaaaatcaa     2280

ggctacaaac taacttttga aaatatatca gagagctata ttgatagcgt agttaatcag     2340

ggtaaattgt acctattcca aatctataat aaagattttt cagcttatag caaagggcga     2400

ccaaatctac atactttata ttggaaagcg ctgtttgatg agagaaatct tcaagatgtg     2460

gtttataagc taaatggtga ggcagagctt ttttatcgta aacaatcaat acctaaaaaa     2520

atcactcacc cagctaaaga ggcaatagct aataaaaaca aagataatcc taaaaaagag     2580

agtgtttttg aatatgattt aatcaaagat aaacgcttta ctgaagataa gtttttcttt     2640

cactgtccta ttacaatcaa ttttaaatct agtggagcta ataagtttaa tgatgaaatc     2700

aatttattgc taaaagaaaa agcaaatgat gttcatatat taagtataga tagaggtgaa     2760

agacatttag cttactatac tttggtagat ggtaaaggca atatcatcaa acaagatact     2820

ttcaacatca ttggtaatga tagaatgaaa acaaactacc atgataagct tgctgcaata     2880

gagaaagata gggattcagc taggaaagac tggaaaaaga taaataacat caaagagatg     2940

aaagagggct atctatctca ggtagttcat gaaatagcta agctagttat agagtataat     3000

gctattgtgg tttttgagga tttaaatttt ggatttaaaa gagggcgttt caaggtagag     3060

aagcaggtct atcaaaagtt agaaaaaatg ctaattgaga aactaaacta tctagttttc     3120

aaagataatg agtttgataa aactggggga gtgcttagag cttatcagct aacagcacct     3180

tttgagactt ttaaaaagat gggtaaacaa acaggtatta tctactatgt accagctggt     3240

tttacttcaa aaatttgtcc tgtaactggt tttgtaaatc agttatatcc taagtatgaa     3300

agtgtcagca aatctcaaga gttctttagt aagtttgaca agatttgtta taaccttgat     3360

aagggctatt ttgagtttag ttttgattat aaaaactttg gtgacaaggc tgccaaaggc     3420

aagtggacta tagctagctt tgggagtaga ttgattaact ttagaaattc agataaaaat     3480

cataattggg atactcgaga agtttatcca actaaagagt tggagaaatt gctaaaagat     3540

tattctatcg aatatgggca tggcgaatgt atcaaagcag ctatttgcgg tgagagcgac     3600

aaaaagtttt ttgctaagct aactagtgtc ctaaatacta tcttacaaat gcgtaactca     3660

aaaacaggta ctgagttaga ttatctaatt tcaccagtag cagatgtaaa tggcaatttc     3720

tttgattcgc gacaggcgcc aaaaaatatg cctcaagatg ctgatgccaa tggtgcttat     3780

catattgggc taaaaggtct gatgctacta ggtaggatca aaaataatca agagggcaaa     3840

aaactcaatt tggttatcaa aaatgaagag tattttgagt tcgtgcagaa taggaataac     3900

taattcattc aagaatatat taccctgtca gtttagcgac tattacctct ttaataattt     3960


<210>  9
<211>  2092
<212>  PRT
<213>  Artificial

<220>
<223>  PE2 (prime editor)


<220>
<221>  misc_feature
<222>  (1)..(2092)
<223>  PE2

<400>  9

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 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             


Ser Gly  Gly Ser Ser Gly Gly  Ser Ser Gly Ser Glu  Thr Pro Gly 
    1370                 1375                 1380             


Thr Ser  Glu Ser Ala Thr Pro  Glu Ser Ser Gly Gly  Ser Ser Gly 
    1385                 1390                 1395             


Gly Ser  Ser Thr Leu Asn Ile  Glu Asp Glu Tyr Arg  Leu His Glu 
    1400                 1405                 1410             


Thr Ser  Lys Glu Pro Asp Val  Ser Leu Gly Ser Thr  Trp Leu Ser 
    1415                 1420                 1425             


Asp Phe  Pro Gln Ala Trp Ala  Glu Thr Gly Gly Met  Gly Leu Ala 
    1430                 1435                 1440             


Val Arg  Gln Ala Pro Leu Ile  Ile Pro Leu Lys Ala  Thr Ser Thr 
    1445                 1450                 1455             


Pro Val  Ser Ile Lys Gln Tyr  Pro Met Ser Gln Glu  Ala Arg Leu 
    1460                 1465                 1470             


Gly Ile  Lys Pro His Ile Gln  Arg Leu Leu Asp Gln  Gly Ile Leu 
    1475                 1480                 1485             


Val Pro  Cys Gln Ser Pro Trp  Asn Thr Pro Leu Leu  Pro Val Lys 
    1490                 1495                 1500             


Lys Pro  Gly Thr Asn Asp Tyr  Arg Pro Val Gln Asp  Leu Arg Glu 
    1505                 1510                 1515             


Val Asn  Lys Arg Val Glu Asp  Ile His Pro Thr Val  Pro Asn Pro 
    1520                 1525                 1530             


Tyr Asn  Leu Leu Ser Gly Leu  Pro Pro Ser His Gln  Trp Tyr Thr 
    1535                 1540                 1545             


Val Leu  Asp Leu Lys Asp Ala  Phe Phe Cys Leu Arg  Leu His Pro 
    1550                 1555                 1560             


Thr Ser  Gln Pro Leu Phe Ala  Phe Glu Trp Arg Asp  Pro Glu Met 
    1565                 1570                 1575             


Gly Ile  Ser Gly Gln Leu Thr  Trp Thr Arg Leu Pro  Gln Gly Phe 
    1580                 1585                 1590             


Lys Asn  Ser Pro Thr Leu Phe  Asn Glu Ala Leu His  Arg Asp Leu 
    1595                 1600                 1605             


Ala Asp  Phe Arg Ile Gln His  Pro Asp Leu Ile Leu  Leu Gln Tyr 
    1610                 1615                 1620             


Val Asp  Asp Leu Leu Leu Ala  Ala Thr Ser Glu Leu  Asp Cys Gln 
    1625                 1630                 1635             


Gln Gly  Thr Arg Ala Leu Leu  Gln Thr Leu Gly Asn  Leu Gly Tyr 
    1640                 1645                 1650             


Arg Ala  Ser Ala Lys Lys Ala  Gln Ile Cys Gln Lys  Gln Val Lys 
    1655                 1660                 1665             


Tyr Leu  Gly Tyr Leu Leu Lys  Glu Gly Gln Arg Trp  Leu Thr Glu 
    1670                 1675                 1680             


Ala Arg  Lys Glu Thr Val Met  Gly Gln Pro Thr Pro  Lys Thr Pro 
    1685                 1690                 1695             


Arg Gln  Leu Arg Glu Phe Leu  Gly Lys Ala Gly Phe  Cys Arg Leu 
    1700                 1705                 1710             


Phe Ile  Pro Gly Phe Ala Glu  Met Ala Ala Pro Leu  Tyr Pro Leu 
    1715                 1720                 1725             


Thr Lys  Pro Gly Thr Leu Phe  Asn Trp Gly Pro Asp  Gln Gln Lys 
    1730                 1735                 1740             


Ala Tyr  Gln Glu Ile Lys Gln  Ala Leu Leu Thr Ala  Pro Ala Leu 
    1745                 1750                 1755             


Gly Leu  Pro Asp Leu Thr Lys  Pro Phe Glu Leu Phe  Val Asp Glu 
    1760                 1765                 1770             


Lys Gln  Gly Tyr Ala Lys Gly  Val Leu Thr Gln Lys  Leu Gly Pro 
    1775                 1780                 1785             


Trp Arg  Arg Pro Val Ala Tyr  Leu Ser Lys Lys Leu  Asp Pro Val 
    1790                 1795                 1800             


Ala Ala  Gly Trp Pro Pro Cys  Leu Arg Met Val Ala  Ala Ile Ala 
    1805                 1810                 1815             


Val Leu  Thr Lys Asp Ala Gly  Lys Leu Thr Met Gly  Gln Pro Leu 
    1820                 1825                 1830             


Val Ile  Leu Ala Pro His Ala  Val Glu Ala Leu Val  Lys Gln Pro 
    1835                 1840                 1845             


Pro Asp  Arg Trp Leu Ser Asn  Ala Arg Met Thr His  Tyr Gln Ala 
    1850                 1855                 1860             


Leu Leu  Leu Asp Thr Asp Arg  Val Gln Phe Gly Pro  Val Val Ala 
    1865                 1870                 1875             


Leu Asn  Pro Ala Thr Leu Leu  Pro Leu Pro Glu Glu  Gly Leu Gln 
    1880                 1885                 1890             


His Asn  Cys Leu Asp Ile Leu  Ala Glu Ala His Gly  Thr Arg Pro 
    1895                 1900                 1905             


Asp Leu  Thr Asp Gln Pro Leu  Pro Asp Ala Asp His  Thr Trp Tyr 
    1910                 1915                 1920             


Thr Asp  Gly Ser Ser Leu Leu  Gln Glu Gly Gln Arg  Lys Ala Gly 
    1925                 1930                 1935             


Ala Ala  Val Thr Thr Glu Thr  Glu Val Ile Trp Ala  Lys Ala Leu 
    1940                 1945                 1950             


Pro Ala  Gly Thr Ser Ala Gln  Arg Ala Glu Leu Ile  Ala Leu Thr 
    1955                 1960                 1965             


Gln Ala  Leu Lys Met Ala Glu  Gly Lys Lys Leu Asn  Val Tyr Thr 
    1970                 1975                 1980             


Asp Ser  Arg Tyr Ala Phe Ala  Thr Ala His Ile His  Gly Glu Ile 
    1985                 1990                 1995             


Tyr Arg  Arg Arg Gly Trp Leu  Thr Ser Glu Gly Lys  Glu Ile Lys 
    2000                 2005                 2010             


Asn Lys  Asp Glu Ile Leu Ala  Leu Leu Lys Ala Leu  Phe Leu Pro 
    2015                 2020                 2025             


Lys Arg  Leu Ser Ile Ile His  Cys Pro Gly His Gln  Lys Gly His 
    2030                 2035                 2040             


Ser Ala  Glu Ala Arg Gly Asn  Arg Met Ala Asp Gln  Ala Ala Arg 
    2045                 2050                 2055             


Lys Ala  Ala Ile Thr Glu Thr  Pro Asp Thr Ser Thr  Leu Leu Ile 
    2060                 2065                 2070             


Glu Asn  Ser Ser Pro Ser Gly  Gly Ser Lys Arg Thr  Ala Asp Gly 
    2075                 2080                 2085             


Ser Glu  Phe Glu 
    2090         


<210>  10
<211>  6279
<212>  DNA
<213>  Artificial

<220>
<223>  PE2 (primer editor) DNA


<220>
<221>  misc_feature
<222>  (1)..(6279)
<223>  PE2 (DNA)

<400>  10
atggacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg      120

cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag      180

gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc      240

tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga      300

ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc      360

aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag      420

aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac      540

gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc      600

atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga      660

cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac      720

ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag      780

gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc      840

cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc      900

ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct      960

atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg     1020

cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc     1080

ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg     1140

gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg     1200

aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac     1260

gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc     1320

gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc     1380

agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa     1440

gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag     1500

aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg     1560

tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg     1620

agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc     1680

gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc     1740

tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt     1800

atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg     1860

ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc     1920

cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc     1980

aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg     2040

gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac     2100

agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg     2160

cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca     2220

gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg     2280

atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga     2340

atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc     2400

gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg     2460

gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct     2520

atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc     2580

gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag     2640

aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg     2700

accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag     2760

ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac     2820

actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc     2880

aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac     2940

taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag     3000

taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag     3060

atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc     3120

aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg     3180

cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt     3240

gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg     3300

cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc     3360

gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc     3420

tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg     3480

aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac     3540

tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag     3600

tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg     3660

cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc     3720

cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa     3780

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag     3900

cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc     3960

cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa     4020

gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg tgactctgga ggatctagcg gaggatcctc tggcagcgag     4140

acaccaggaa caagcgagtc agcaacacca gagagcagtg gcggcagcag cggcggcagc     4200

agcaccctaa atatagaaga tgagtatcgg ctacatgaga cctcaaaaga gccagatgtt     4260

tctctagggt ccacatggct gtctgatttt cctcaggcct gggcggaaac cgggggcatg     4320

ggactggcag ttcgccaagc tcctctgatc atacctctga aagcaacctc tacccccgtg     4380

tccataaaac aataccccat gtcacaagaa gccagactgg ggatcaagcc ccacatacag     4440

agactgttgg accagggaat actggtaccc tgccagtccc cctggaacac gcccctgcta     4500

cccgttaaga aaccagggac taatgattat aggcctgtcc aggatctgag agaagtcaac     4560

aagcgggtgg aagacatcca ccccaccgtg cccaaccctt acaacctctt gagcgggctc     4620

ccaccgtccc accagtggta cactgtgctt gatttaaagg atgccttttt ctgcctgaga     4680

ctccacccca ccagtcagcc tctcttcgcc tttgagtgga gagatccaga gatgggaatc     4740

tcaggacaat tgacctggac cagactccca cagggtttca aaaacagtcc caccctgttt     4800

aatgaggcac tgcacagaga cctagcagac ttccggatcc agcacccaga cttgatcctg     4860

ctacagtacg tggatgactt actgctggcc gccacttctg agctagactg ccaacaaggt     4920

actcgggccc tgttacaaac cctagggaac ctcgggtatc gggcctcggc caagaaagcc     4980

caaatttgcc agaaacaggt caagtatctg gggtatcttc taaaagaggg tcagagatgg     5040

ctgactgagg ccagaaaaga gactgtgatg gggcagccta ctccgaagac ccctcgacaa     5100

ctaagggagt tcctagggaa ggcaggcttc tgtcgcctct tcatccctgg gtttgcagaa     5160

atggcagccc ccctgtaccc tctcaccaaa ccggggactc tgtttaattg gggcccagac     5220

caacaaaagg cctatcaaga aatcaagcaa gctcttctaa ctgccccagc cctggggttg     5280

ccagatttga ctaagccctt tgaactcttt gtcgacgaga agcagggcta cgccaaaggt     5340

gtcctaacgc aaaaactggg accttggcgt cggccggtgg cctacctgtc caaaaagcta     5400

gacccagtag cagctgggtg gcccccttgc ctacggatgg tagcagccat tgccgtactg     5460

acaaaggatg caggcaagct aaccatggga cagccactag tcattctggc cccccatgca     5520

gtagaggcac tagtcaaaca accccccgac cgctggcttt ccaacgcccg gatgactcac     5580

tatcaggcct tgcttttgga cacggaccgg gtccagttcg gaccggtggt agccctgaac     5640

ccggctacgc tgctcccact gcctgaggaa gggctgcaac acaactgcct tgatatcctg     5700

gccgaagccc acggaacccg acccgaccta acggaccagc cgctcccaga cgccgaccac     5760

acctggtaca cggatggaag cagtctctta caagagggac agcgtaaggc gggagctgcg     5820

gtgaccaccg agaccgaggt aatctgggct aaagccctgc cagccgggac atccgctcag     5880

cgggctgaac tgatagcact cacccaggcc ctaaagatgg cagaaggtaa gaagctaaat     5940

gtttatactg atagccgtta tgcttttgct actgcccata tccatggaga aatatacaga     6000

aggcgtgggt ggctcacatc agaaggcaaa gagatcaaaa ataaagacga gatcttggcc     6060

ctactaaaag ccctctttct gcccaaaaga cttagcataa tccattgtcc aggacatcaa     6120

aagggacaca gcgccgaggc tagaggcaac cggatggctg accaagcggc ccgaaaggca     6180

gccatcacag agactccaga cacctctacc ctcctcatag aaaattcatc accctctggc     6240

ggctcaaaaa gaaccgccga cggcagcgaa ttcgagtaa                            6279


