                         SEQUENCE LISTING

<110>  EmendoBio Inc.
       IZHAR, Lior
       DIAMANT, Noam
       ZILBERMAN, Yuliya
 
<120>  NOVEL GENOME EDITING TOOL

<130>  91004-A-PCT/GJG/AWG

<150>  62/860,629
<151>  2019-06-12

<150>  63/029,679
<151>  2020-05-25

<160>  72    

<170>  PatentIn version 3.5

<210>  1
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 1 - BE4 short linker

<400>  1

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


<210>  2
<211>  32
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 2 - BE4 long linker

<400>  2

Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly Ser Glu Thr Pro Gly Thr 
1               5                   10                  15      


Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly Gly Ser Ser Gly Gly Ser 
            20                  25                  30          


<210>  3
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 3 - Dead SpCas9

<400>  3

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  4
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 4 - D10A Nickase SpCas9

<400>  4

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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

<220>
<223>  SEQ ID NO: 5 - H840A SpCas9 nickase

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  6
<211>  1114
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 6 - Bm R2 CDS

<400>  6

Met Met Ala Ser Thr Ala Leu Ser Leu Met Gly Arg Cys Asn Pro Asp 
1               5                   10                  15      


Gly Cys Thr Arg Gly Lys His Val Thr Ala Ala Pro Met Asp Gly Pro 
            20                  25                  30          


Arg Gly Pro Ser Ser Leu Ala Gly Thr Phe Gly Trp Gly Leu Ala Ile 
        35                  40                  45              


Pro Ala Gly Glu Pro Cys Gly Arg Val Cys Ser Pro Ala Thr Val Gly 
    50                  55                  60                  


Phe Phe Pro Val Ala Lys Lys Ser Asn Lys Glu Asn Arg Pro Glu Ala 
65                  70                  75                  80  


Ser Gly Leu Pro Leu Glu Ser Glu Arg Thr Gly Asp Asn Pro Thr Val 
                85                  90                  95      


Arg Gly Ser Ala Gly Ala Asp Pro Val Gly Gln Asp Ala Pro Gly Trp 
            100                 105                 110         


Thr Cys Gln Phe Cys Glu Arg Thr Phe Ser Thr Asn Arg Gly Leu Gly 
        115                 120                 125             


Val His Lys Arg Arg Ala His Pro Val Glu Thr Asn Thr Asp Ala Ala 
    130                 135                 140                 


Pro Met Met Val Lys Arg Arg Trp His Gly Glu Glu Ile Asp Leu Leu 
145                 150                 155                 160 


Ala Arg Thr Glu Ala Arg Leu Leu Ala Glu Arg Gly Gln Cys Ser Gly 
                165                 170                 175     


Gly Asp Leu Phe Gly Ala Leu Pro Gly Phe Gly Arg Thr Leu Glu Ala 
            180                 185                 190         


Ile Lys Gly Gln Arg Arg Arg Glu Pro Tyr Arg Ala Leu Val Gln Ala 
        195                 200                 205             


His Leu Ala Arg Phe Gly Ser Gln Pro Gly Pro Ser Ser Gly Gly Cys 
    210                 215                 220                 


Ser Ala Glu Pro Asp Phe Arg Arg Ala Ser Gly Ala Glu Glu Ala Gly 
225                 230                 235                 240 


Glu Glu Arg Cys Ala Glu Asp Ala Ala Ala Tyr Asp Pro Ser Ala Val 
                245                 250                 255     


Gly Gln Met Ser Pro Asp Ala Ala Arg Val Leu Ser Glu Leu Leu Glu 
            260                 265                 270         


Gly Ala Gly Arg Arg Arg Ala Cys Arg Ala Met Arg Pro Lys Thr Ala 
        275                 280                 285             


Gly Arg Arg Asn Asp Leu His Asp Asp Arg Thr Ala Ser Ala His Lys 
    290                 295                 300                 


Thr Ser Arg Gln Lys Arg Arg Ala Glu Tyr Ala Arg Val Gln Glu Leu 
305                 310                 315                 320 


Tyr Lys Lys Cys Arg Ser Arg Ala Ala Ala Glu Val Ile Asp Gly Ala 
                325                 330                 335     


Cys Gly Gly Val Gly His Ser Leu Glu Glu Met Glu Thr Tyr Trp Arg 
            340                 345                 350         


Pro Ile Leu Glu Arg Val Ser Asp Ala Pro Gly Pro Thr Pro Glu Ala 
        355                 360                 365             


Leu His Ala Leu Gly Arg Ala Glu Trp His Gly Gly Asn Arg Asp Tyr 
    370                 375                 380                 


Thr Gln Leu Trp Lys Pro Ile Ser Val Glu Glu Ile Lys Ala Ser Arg 
385                 390                 395                 400 


Phe Asp Trp Arg Thr Ser Pro Gly Pro Asp Gly Ile Arg Ser Gly Gln 
                405                 410                 415     


Trp Arg Ala Val Pro Val His Leu Lys Ala Glu Met Phe Asn Ala Trp 
            420                 425                 430         


Met Ala Arg Gly Glu Ile Pro Glu Ile Leu Arg Gln Cys Arg Thr Val 
        435                 440                 445             


Phe Val Pro Lys Val Glu Arg Pro Gly Gly Pro Gly Glu Tyr Arg Pro 
    450                 455                 460                 


Ile Ser Ile Ala Ser Ile Pro Leu Arg His Phe His Ser Ile Leu Ala 
465                 470                 475                 480 


Arg Arg Leu Leu Ala Cys Cys Pro Pro Asp Ala Arg Gln Arg Gly Phe 
                485                 490                 495     


Ile Cys Ala Asp Gly Thr Leu Glu Asn Ser Ala Val Leu Asp Ala Val 
            500                 505                 510         


Leu Gly Asp Ser Arg Lys Lys Leu Arg Glu Cys His Val Ala Val Leu 
        515                 520                 525             


Asp Phe Ala Lys Ala Phe Asp Thr Val Ser His Glu Ala Leu Val Glu 
    530                 535                 540                 


Leu Leu Arg Leu Arg Gly Met Pro Glu Gln Phe Cys Gly Tyr Ile Ala 
545                 550                 555                 560 


His Leu Tyr Asp Thr Ala Ser Thr Thr Leu Ala Val Asn Asn Glu Met 
                565                 570                 575     


Ser Ser Pro Val Lys Val Gly Arg Gly Val Arg Gln Gly Asp Pro Leu 
            580                 585                 590         


Ser Pro Ile Leu Phe Asn Val Val Met Asp Leu Ile Leu Ala Ser Leu 
        595                 600                 605             


Pro Glu Arg Val Gly Tyr Arg Leu Glu Met Glu Leu Val Ser Ala Leu 
    610                 615                 620                 


Ala Tyr Ala Asp Asp Leu Val Leu Leu Ala Gly Ser Lys Val Gly Met 
625                 630                 635                 640 


Gln Glu Ser Ile Ser Ala Val Asp Cys Val Gly Arg Gln Met Gly Leu 
                645                 650                 655     


Arg Leu Asn Cys Arg Lys Ser Ala Val Leu Ser Met Ile Pro Asp Gly 
            660                 665                 670         


His Arg Lys Lys His His Tyr Leu Thr Glu Arg Thr Phe Asn Ile Gly 
        675                 680                 685             


Gly Lys Pro Leu Arg Gln Val Ser Cys Val Glu Arg Trp Arg Tyr Leu 
    690                 695                 700                 


Gly Val Asp Phe Glu Ala Ser Gly Cys Val Thr Leu Glu His Ser Ile 
705                 710                 715                 720 


Ser Ser Ala Leu Asn Asn Ile Ser Arg Ala Pro Leu Lys Pro Gln Gln 
                725                 730                 735     


Arg Leu Glu Ile Leu Arg Ala His Leu Ile Pro Arg Phe Gln His Gly 
            740                 745                 750         


Phe Val Leu Gly Asn Ile Ser Asp Asp Arg Leu Arg Met Leu Asp Val 
        755                 760                 765             


Gln Ile Arg Lys Ala Val Gly Gln Trp Leu Arg Leu Pro Ala Asp Val 
    770                 775                 780                 


Pro Lys Ala Tyr Tyr His Ala Ala Val Gln Asp Gly Gly Leu Ala Ile 
785                 790                 795                 800 


Pro Ser Val Arg Ala Thr Ile Pro Asp Leu Ile Val Arg Arg Phe Gly 
                805                 810                 815     


Gly Leu Asp Ser Ser Pro Trp Ser Val Ala Arg Ala Ala Ala Lys Ser 
            820                 825                 830         


Asp Lys Ile Arg Lys Lys Leu Arg Trp Ala Trp Lys Gln Leu Arg Arg 
        835                 840                 845             


Phe Ser Arg Val Asp Ser Thr Thr Gln Arg Pro Ser Val Arg Leu Phe 
    850                 855                 860                 


Trp Arg Glu His Leu His Ala Ser Val Asp Gly Arg Glu Leu Arg Glu 
865                 870                 875                 880 


Ser Thr Arg Thr Pro Thr Ser Thr Lys Trp Ile Arg Glu Arg Cys Ala 
                885                 890                 895     


Gln Ile Thr Gly Arg Asp Phe Val Gln Phe Val His Thr His Ile Asn 
            900                 905                 910         


Ala Leu Pro Ser Arg Ile Arg Gly Ser Arg Gly Arg Arg Gly Gly Gly 
        915                 920                 925             


Glu Ser Ser Leu Thr Cys Arg Ala Gly Cys Lys Val Arg Glu Thr Thr 
    930                 935                 940                 


Ala His Ile Leu Gln Gln Cys His Arg Thr His Gly Gly Arg Ile Leu 
945                 950                 955                 960 


Arg His Asn Lys Ile Val Ser Phe Val Ala Lys Ala Met Glu Glu Asn 
                965                 970                 975     


Lys Trp Thr Val Glu Leu Glu Pro Arg Leu Arg Thr Ser Val Gly Leu 
            980                 985                 990         


Arg Lys Pro Asp Ile Ile Ala Ser  Arg Asp Gly Val Gly  Val Ile Val 
        995                 1000                 1005             


Asp Val  Gln Val Val Ser Gly  Gln Arg Ser Leu Asp  Glu Leu His 
    1010                 1015                 1020             


Arg Glu  Lys Arg Asn Lys Tyr  Gly Asn His Gly Glu  Leu Val Glu 
    1025                 1030                 1035             


Leu Val  Ala Gly Arg Leu Gly  Leu Pro Lys Ala Glu  Cys Val Arg 
    1040                 1045                 1050             


Ala Thr  Ser Cys Thr Ile Ser  Trp Arg Gly Val Trp  Ser Leu Thr 
    1055                 1060                 1065             


Ser Tyr  Lys Glu Leu Arg Ser  Ile Ile Gly Leu Arg  Glu Pro Thr 
    1070                 1075                 1080             


Leu Gln  Ile Val Pro Ile Leu  Ala Leu Arg Gly Ser  His Met Asn 
    1085                 1090                 1095             


Trp Thr  Arg Phe Asn Gln Met  Thr Ser Val Met Gly  Gly Gly Val 
    1100                 1105                 1110             


Gly 
    


<210>  7
<211>  75
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 7 - 5' R2 RNA pseudoknot

<400>  7
gccccgatgg acggaccgcg aggaccgtca agcctagcag gtaccttcgg gtggggcctt       60

gcgatacctg cgggc                                                        75


<210>  8
<211>  248
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 8 - 3' R2 RNA structured region

<400>  8
gccttgcaca gtagtccagc ggtaagggtg tagatcaggc ccgtctgttt ctcccccgga       60

gctcgctccc ttggcttccc ttatatattt taacatcaga aacagacatt aaacatctac      120

tgatccaatt tcgccggcgt acggccacga tcgggagggt gggaatctcg ggggtcttcc      180

gatcctaatc catgatgatt acgacctgag tcactaaaga cgatggcatg atgatccggc      240

gatgaaaa                                                               248


<210>  9
<211>  398
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 9 - R2 RNA BD + RT domain

<400>  9

Arg Ala Glu Tyr Ala Arg Val Gln Glu Leu Tyr Lys Lys Cys Arg Ser 
1               5                   10                  15      


Arg Ala Ala Ala Glu Val Ile Asp Gly Ala Cys Gly Gly Val Gly His 
            20                  25                  30          


Ser Leu Glu Glu Met Glu Thr Tyr Trp Arg Pro Ile Leu Glu Arg Val 
        35                  40                  45              


Ser Asp Ala Pro Gly Pro Thr Pro Glu Ala Leu His Ala Leu Gly Arg 
    50                  55                  60                  


Ala Glu Trp His Gly Gly Asn Arg Asp Tyr Thr Gln Leu Trp Lys Pro 
65                  70                  75                  80  


Ile Ser Val Glu Glu Ile Lys Ala Ser Arg Phe Asp Trp Arg Thr Ser 
                85                  90                  95      


Pro Gly Pro Asp Gly Ile Arg Ser Gly Gln Trp Arg Ala Val Pro Val 
            100                 105                 110         


His Leu Lys Ala Glu Met Phe Asn Ala Trp Met Ala Arg Gly Glu Ile 
        115                 120                 125             


Pro Glu Ile Leu Arg Gln Cys Arg Thr Val Phe Val Pro Lys Val Glu 
    130                 135                 140                 


Arg Pro Gly Gly Pro Gly Glu Tyr Arg Pro Ile Ser Ile Ala Ser Ile 
145                 150                 155                 160 


Pro Leu Arg His Phe His Ser Ile Leu Ala Arg Arg Leu Leu Ala Cys 
                165                 170                 175     


Cys Pro Pro Asp Ala Arg Gln Arg Gly Phe Ile Cys Ala Asp Gly Thr 
            180                 185                 190         


Leu Glu Asn Ser Ala Val Leu Asp Ala Val Leu Gly Asp Ser Arg Lys 
        195                 200                 205             


Lys Leu Arg Glu Cys His Val Ala Val Leu Asp Phe Ala Lys Ala Phe 
    210                 215                 220                 


Asp Thr Val Ser His Glu Ala Leu Val Glu Leu Leu Arg Leu Arg Gly 
225                 230                 235                 240 


Met Pro Glu Gln Phe Cys Gly Tyr Ile Ala His Leu Tyr Asp Thr Ala 
                245                 250                 255     


Ser Thr Thr Leu Ala Val Asn Asn Glu Met Ser Ser Pro Val Lys Val 
            260                 265                 270         


Gly Arg Gly Val Arg Gln Gly Asp Pro Leu Ser Pro Ile Leu Phe Asn 
        275                 280                 285             


Val Val Met Asp Leu Ile Leu Ala Ser Leu Pro Glu Arg Val Gly Tyr 
    290                 295                 300                 


Arg Leu Glu Met Glu Leu Val Ser Ala Leu Ala Tyr Ala Asp Asp Leu 
305                 310                 315                 320 


Val Leu Leu Ala Gly Ser Lys Val Gly Met Gln Glu Ser Ile Ser Ala 
                325                 330                 335     


Val Asp Cys Val Gly Arg Gln Met Gly Leu Arg Leu Asn Cys Arg Lys 
            340                 345                 350         


Ser Ala Val Leu Ser Met Ile Pro Asp Gly His Arg Lys Lys His His 
        355                 360                 365             


Tyr Leu Thr Glu Arg Thr Phe Asn Ile Gly Gly Lys Pro Leu Arg Gln 
    370                 375                 380                 


Val Ser Cys Val Glu Arg Trp Arg Tyr Leu Gly Val Asp Phe 
385                 390                 395             


<210>  10
<211>  803
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 10 - R2 RNA BD + RT domain + Endonuclease domain

<400>  10

Arg Ala Glu Tyr Ala Arg Val Gln Glu Leu Tyr Lys Lys Cys Arg Ser 
1               5                   10                  15      


Arg Ala Ala Ala Glu Val Ile Asp Gly Ala Cys Gly Gly Val Gly His 
            20                  25                  30          


Ser Leu Glu Glu Met Glu Thr Tyr Trp Arg Pro Ile Leu Glu Arg Val 
        35                  40                  45              


Ser Asp Ala Pro Gly Pro Thr Pro Glu Ala Leu His Ala Leu Gly Arg 
    50                  55                  60                  


Ala Glu Trp His Gly Gly Asn Arg Asp Tyr Thr Gln Leu Trp Lys Pro 
65                  70                  75                  80  


Ile Ser Val Glu Glu Ile Lys Ala Ser Arg Phe Asp Trp Arg Thr Ser 
                85                  90                  95      


Pro Gly Pro Asp Gly Ile Arg Ser Gly Gln Trp Arg Ala Val Pro Val 
            100                 105                 110         


His Leu Lys Ala Glu Met Phe Asn Ala Trp Met Ala Arg Gly Glu Ile 
        115                 120                 125             


Pro Glu Ile Leu Arg Gln Cys Arg Thr Val Phe Val Pro Lys Val Glu 
    130                 135                 140                 


Arg Pro Gly Gly Pro Gly Glu Tyr Arg Pro Ile Ser Ile Ala Ser Ile 
145                 150                 155                 160 


Pro Leu Arg His Phe His Ser Ile Leu Ala Arg Arg Leu Leu Ala Cys 
                165                 170                 175     


Cys Pro Pro Asp Ala Arg Gln Arg Gly Phe Ile Cys Ala Asp Gly Thr 
            180                 185                 190         


Leu Glu Asn Ser Ala Val Leu Asp Ala Val Leu Gly Asp Ser Arg Lys 
        195                 200                 205             


Lys Leu Arg Glu Cys His Val Ala Val Leu Asp Phe Ala Lys Ala Phe 
    210                 215                 220                 


Asp Thr Val Ser His Glu Ala Leu Val Glu Leu Leu Arg Leu Arg Gly 
225                 230                 235                 240 


Met Pro Glu Gln Phe Cys Gly Tyr Ile Ala His Leu Tyr Asp Thr Ala 
                245                 250                 255     


Ser Thr Thr Leu Ala Val Asn Asn Glu Met Ser Ser Pro Val Lys Val 
            260                 265                 270         


Gly Arg Gly Val Arg Gln Gly Asp Pro Leu Ser Pro Ile Leu Phe Asn 
        275                 280                 285             


Val Val Met Asp Leu Ile Leu Ala Ser Leu Pro Glu Arg Val Gly Tyr 
    290                 295                 300                 


Arg Leu Glu Met Glu Leu Val Ser Ala Leu Ala Tyr Ala Asp Asp Leu 
305                 310                 315                 320 


Val Leu Leu Ala Gly Ser Lys Val Gly Met Gln Glu Ser Ile Ser Ala 
                325                 330                 335     


Val Asp Cys Val Gly Arg Gln Met Gly Leu Arg Leu Asn Cys Arg Lys 
            340                 345                 350         


Ser Ala Val Leu Ser Met Ile Pro Asp Gly His Arg Lys Lys His His 
        355                 360                 365             


Tyr Leu Thr Glu Arg Thr Phe Asn Ile Gly Gly Lys Pro Leu Arg Gln 
    370                 375                 380                 


Val Ser Cys Val Glu Arg Trp Arg Tyr Leu Gly Val Asp Phe Glu Ala 
385                 390                 395                 400 


Ser Gly Cys Val Thr Leu Glu His Ser Ile Ser Ser Ala Leu Asn Asn 
                405                 410                 415     


Ile Ser Arg Ala Pro Leu Lys Pro Gln Gln Arg Leu Glu Ile Leu Arg 
            420                 425                 430         


Ala His Leu Ile Pro Arg Phe Gln His Gly Phe Val Leu Gly Asn Ile 
        435                 440                 445             


Ser Asp Asp Arg Leu Arg Met Leu Asp Val Gln Ile Arg Lys Ala Val 
    450                 455                 460                 


Gly Gln Trp Leu Arg Leu Pro Ala Asp Val Pro Lys Ala Tyr Tyr His 
465                 470                 475                 480 


Ala Ala Val Gln Asp Gly Gly Leu Ala Ile Pro Ser Val Arg Ala Thr 
                485                 490                 495     


Ile Pro Asp Leu Ile Val Arg Arg Phe Gly Gly Leu Asp Ser Ser Pro 
            500                 505                 510         


Trp Ser Val Ala Arg Ala Ala Ala Lys Ser Asp Lys Ile Arg Lys Lys 
        515                 520                 525             


Leu Arg Trp Ala Trp Lys Gln Leu Arg Arg Phe Ser Arg Val Asp Ser 
    530                 535                 540                 


Thr Thr Gln Arg Pro Ser Val Arg Leu Phe Trp Arg Glu His Leu His 
545                 550                 555                 560 


Ala Ser Val Asp Gly Arg Glu Leu Arg Glu Ser Thr Arg Thr Pro Thr 
                565                 570                 575     


Ser Thr Lys Trp Ile Arg Glu Arg Cys Ala Gln Ile Thr Gly Arg Asp 
            580                 585                 590         


Phe Val Gln Phe Val His Thr His Ile Asn Ala Leu Pro Ser Arg Ile 
        595                 600                 605             


Arg Gly Ser Arg Gly Arg Arg Gly Gly Gly Glu Ser Ser Leu Thr Cys 
    610                 615                 620                 


Arg Ala Gly Cys Lys Val Arg Glu Thr Thr Ala His Ile Leu Gln Gln 
625                 630                 635                 640 


Cys His Arg Thr His Gly Gly Arg Ile Leu Arg His Asn Lys Ile Val 
                645                 650                 655     


Ser Phe Val Ala Lys Ala Met Glu Glu Asn Lys Trp Thr Val Glu Leu 
            660                 665                 670         


Glu Pro Arg Leu Arg Thr Ser Val Gly Leu Arg Lys Pro Asp Ile Ile 
        675                 680                 685             


Ala Ser Arg Asp Gly Val Gly Val Ile Val Asp Val Gln Val Val Ser 
    690                 695                 700                 


Gly Gln Arg Ser Leu Asp Glu Leu His Arg Glu Lys Arg Asn Lys Tyr 
705                 710                 715                 720 


Gly Asn His Gly Glu Leu Val Glu Leu Val Ala Gly Arg Leu Gly Leu 
                725                 730                 735     


Pro Lys Ala Glu Cys Val Arg Ala Thr Ser Cys Thr Ile Ser Trp Arg 
            740                 745                 750         


Gly Val Trp Ser Leu Thr Ser Tyr Lys Glu Leu Arg Ser Ile Ile Gly 
        755                 760                 765             


Leu Arg Glu Pro Thr Leu Gln Ile Val Pro Ile Leu Ala Leu Arg Gly 
    770                 775                 780                 


Ser His Met Asn Trp Thr Arg Phe Asn Gln Met Thr Ser Val Met Gly 
785                 790                 795                 800 


Gly Gly Val 
            


<210>  11
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 11 - Forward Primer 5431

<400>  11
tcgggttgct ctcatccctg                                                   20


<210>  12
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 12 - Reverse Primer 5222

<400>  12
cctctcatgt ctcttcaccg tgc                                               23


<210>  13
<211>  5363
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 13 - pcDNA3 backbone sequence

<400>  13
gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg       60

ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg      120

cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc      180

ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt      240

gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata      300

tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc      360

cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc      420

attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt      480

atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt      540

atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca      600

tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg      660

actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc      720

aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg      780

gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca      840

ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc      900

gtttaaactt aagcttctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac      960

tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct     1020

ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct     1080

gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg     1140

ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag     1200

aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc     1260

gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc     1320

tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa     1380

tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact     1440

tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt     1500

gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa     1560

ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt     1620

aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag     1680

ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc     1740

aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa     1800

agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc     1860

ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat     1920

gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt     1980

ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat     2040

caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct     2100

ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc     2160

tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc     2220

gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc     2280

acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg     2340

ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag     2400

aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc     2460

ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt     2520

cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc     2580

gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc     2640

tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg     2700

ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag     2760

cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg     2820

cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg     2880

aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct     2940

tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc     3000

gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg     3060

gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt     3120

ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct     3180

ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc     3240

tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat     3300

gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc     3360

tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg     3420

ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag     3480

cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag     3540

gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc     3600

tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc     3660

agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc     3720

tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt     3780

cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg     3840

ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat     3900

ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag     3960

ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt     4020

ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc     4080

cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta     4140

gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag     4200

atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga     4260

ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa     4320

gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa     4380

tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc     4440

ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga     4500

taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa     4560

gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt     4620

gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg     4680

ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc     4740

aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg     4800

gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag     4860

cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt     4920

actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt     4980

caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac     5040

gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac     5100

ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag     5160

caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa     5220

tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga     5280

gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc     5340

cccgaaaagt gccacctgac gtc                                             5363


<210>  14
<211>  36
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 14 - Kozak-NLS sequence

<400>  14
gccaccatgc ctaagaagaa gagaaaggtg ggtacc                                 36


<210>  15
<211>  3828
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 15 - R2OI protein sequence ORF, codon optimized

<400>  15
atgggcaccg acacagtgta cgtcggccag gattatccta gcggcctgag caaaagagtg       60

cccgctagac tggttgctgg ccccatgctg agagagagat cttgtcacgc ccacgtgttc      120

agagccggac acatgtggaa ttggagaacc agcctgccta gcggcagatg ggatcagcct      180

gctctggaaa agtcccgggt gctgaccaga tctgtggcca ccgctacaga ccccgagatc      240

acatcttacc ctggcaagag cgtgtccacc agcacacagg tgcaagaaga ggactggtgt      300

agcagagaga gcggctggat ttctcctgga ctggcccctg aggaacctag cgtggtgtct      360

gagatcacag cctccatggt ggccactatg agagtggcta cagaggaagt ggtgctggaa      420

cctcagcctg agcaggtcgt gacaattctg cccgagcacg gcagaaatgt gccaccagga      480

ctggccgagc aggataccgc ctctcctatt gaagtgtccg tgctgctgcc cgacctggcc      540

gaaaattgtc ctctgtgtgg tgttcccagc ggcggactga gactgctggg aaagcacttt      600

gccgttagac atgccggcgt gcccgtgacc tacgagtgta gaaagtgtgc ctggcggagc      660

cccaatagcc acagcatctc ttgccacgtg ccaaagtgca gaggcagagc cagaatgcca      720

agcggagatc ctggaatcgc ctgcgatctg tgcgaggcca gatttgccac agaagtggga      780

gtcgcccagc acaagagaca cgtgcacccc gtggaatgga acaaagtgcg gctggaaaga      840

agaggcgcca gaggcggagg aatcaaggcc acaaaacttt ggagcgtggc cgaggtggaa      900

accctgatca gactgattag agagcacggc gatagcggcg ccacatacca gctgattgcc      960

gatgaactcg gcagaggcaa gacagccgag caagtgcgga gcaagaagcg gctgctgaga     1020

atcgataccg ccagcaactc tcccgacgac gccgaagtgg aagaggaaag actggaatct     1080

ctggccgtgc ggtccagcag cagatctcct cctagtctgg tggctaccag agtgcgggaa     1140

gctgtggcaa ggggagaatc tgaaggcggc gaggaaatca gagccattgc cgcactgatc     1200

agagatgtgg atcagaaccc ctgcctgatc gagacaagcg ccagcgacat catcagcaag     1260

ctgggcagaa gagtggacgg ccctaaaaga cccagacctg tcgtgcggga acagacccaa     1320

gaaaaaggct gggtccgacg gctggccaga cggaagagag agtatagaga ggcccagtac     1380

ctgtacagca gagatcaggc aagactggcc gctcagattc tggatggcgc tgcctctcaa     1440

gaatgcgccc tgcctgtgga tcaagtgtac ggcgccttcc gggaaaagtg ggagacagtg     1500

ggacagtttc acggcctggg cgagtttaga acaggcgcta gagccgacaa ctgggagttc     1560

tactctccca tcctggctgc cgaagtcaaa gaaaacctga tgcggatggc caacggcaca     1620

gcccctggac ctgatagaat cagcaagaag gccctgctgg actgggaccc tagaggcgaa     1680

cagctggcta gactgtacac cacatggctg atcggcggcg tgatccccag agtgttcaaa     1740

gagtgtcgga ccaagctgct gcctaagagc agcgatcctg tggaactgca ggatatcgga     1800

ggatggcggc ctgtgacaat cggcagcatg gtcaccagac tgttcagcag aatcctgacc     1860

atgcggctga cccgggcctg tcctatcaat cctagacaga gaggcttcct ggccagcagc     1920

tctggatgtg ccgagaacct gctgatcttc gacgagatcg tgcggcggtc tagaagagat     1980

ggtggaccac tggccgtggt gttcgtggat ttcgccagag ccttcgacag catcagccac     2040

gagcacatcc tgtgtgttct ggaagaaggc ggcctggata gacacgtgat cggcctgatt     2100

cggaacagct acgtggactg tgtgaccaga gtgggctgcg tggaaggcat gacacctcca     2160

atccagatga aggtcggagt gaagcagggc gaccctatga gccctctgct gttcaatctg     2220

gctatggacc ctctgattca caagctggaa acagccggca caggcctgaa gtggggagat     2280

ctgtctatcg ccacactggc cttcgccgat gatctggtgc tggtgtcaga cagcgaagaa     2340

ggcatgggca gatccctggg catcctggaa aaattctgcc agctgaccgg cctgagagtg     2400

cagcctagaa agtgccacgg cttcttcatg gacaagggcg tcgtgaatgg ctgcggcaca     2460

tgggagattt gtggcagccc tatccacatg atcccaccag gcgaatctgt gcgctatctg     2520

ggcgttcaag ttggccctgg aagaggcgtg atggaacccg atctgatccc taccgtgcac     2580

acctggatcg agagaatctc tgaggcccct ctgaagccca gccagagaat gagagtgctg     2640

aatagcttcg ccctgccacg gatcatctat caggctgacc tgggcaaagt gaccgtgaca     2700

aagctggccc agatcgatgg aattgtgcgg aaagccgtga agaagtggct gcatctgagc     2760

cccagcacct gtaatggcct gctgtactcc agaaacagag atggcggact ggggctcctg     2820

aagctggaac gactgattcc tagcgtgcgg accaagagaa tctaccggat gagcagaagc     2880

cccgacatct ggaccagaag aatgaccagc cactccgtgt ccaagagcga ctgggaaatg     2940

ctgtgggtgc aagctggcgg agaaagaggc tctgctcctg ttatgggagc cgtggaagcc     3000

gctcctaccg atgtggaaag atcccctgac taccccgatt ggcggagaga ggaaaatctt     3060

gcttggagcg ccctgagagt tcaaggcgtg ggagctgatc agttcagagg cgatagaacc     3120

tccagcagct ggatcgccga acctgcctct gtgggatttg cccagagaca ttggctggct     3180

gctctggcac ttagagccgg cgtgtaccct accagagagt ttctggccag gggcaaagaa     3240

aagagcggag ccgcctgtag aagatgccct gccagactgg aaagctgcag ccacatcctg     3300

ggccagtgtc ctttcgtgca ggccaacaga atcgcccggc acaacaaagt gtgcgtgctc     3360

ctggcaaccg aggccgagag atttggctgg accgtgatcc gggaattccg gcttgaagat     3420

gctgctggcg ggctgaagat tcccgacctc gtgtgtaaaa aggccgacac cgtgctgatc     3480

gtggacgtga ccgtcagata cgagatggac ggcgagacac tgaagagagc cgccagcgag     3540

aaagtgaagc actatctgcc agtgggccag cagatcaccg acaaagtcgg cggacggtgc     3600

ttcaaagtga tgggctttcc tgtgggcgca agaggcaaat ggccagcctc taacaatacc     3660

gtgctggccg aacttggagt gccagccggc agaatgagga cctttgctag gctggtgtcc     3720

cggcggacac tgctgtatag cctggacatc ctgcgggact tcatgagaga gcctgccgga     3780

agaggtacaa gagtggcact gattccagct gccacaggcg ctgctaac                  3828


<210>  16
<211>  135
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 16 - HA-NLS-P2A sequence

<400>  16
ggatcctacc catacgatgt tccagattac gcggccgctc caaaaaagaa aagaaaagtt       60

gaattcggcg gcagcggcgc caccaacttc agcctgctga agcaggccgg cgacgtggag      120

gagaaccccg gcccc                                                       135


<210>  17
<211>  711
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 17 - mCherry sequence

<400>  17
atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag       60

gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc      120

cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc      180

ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac      240

cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc      300

gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac      360

ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta      420

atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc      480

gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct      540

gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc      600

aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa      660

cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta g               711


<210>  18
<211>  3342
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 18 - R2 protein sequence ORF, codon optimized

<400>  18
atgatggcca gcacagccct gtctctgatg ggcagatgca atcccgatgg ctgcacaaga       60

ggcaagcacg tgacagccgc tcctatggat ggacctagag gaccttctag cctggccggc      120

acatttggat ggggacttgc tattcctgcc ggcgagcctt gtggcagagt gtgttctcct      180

gccaccgtgg gattcttccc agtggccaag aagtccaaca aagagaacag acccgaggcc      240

agcggcctgc ctctggaatc tgaaagaacc ggcgataatc ctaccgtgcg gggatctgct      300

ggtgccgatc ctgttggaca agatgcccct ggctggacct gccagttctg cgagagaacc      360

ttcagcacca atagaggcct gggcgtgcac aaaagacggg ctcaccctgt ggaaacaaac      420

accgacgctg cccctatgat ggtcaagaga agatggcacg gcgaggaaat cgacctgctg      480

gccagaacag aagccagact gctggctgag aggggacagt gttctggcgg agatctgttt      540

ggcgccctgc ctggctttgg aagaaccctg gaagccatca agggccagcg cagaagagag      600

ccttatagag ccctggtgca ggcccacctg gccagatttg gatctcagcc tggacctagc      660

tctggcggat gtagcgccga acctgatttt cggagagcct ctggcgctga agaggccggc      720

gaagaaagat gtgctgagga tgccgccgct tacgatcctt ctgctgtggg ccaaatgagc      780

cctgatgccg ccagagtgct gtctgaactt cttgaaggcg ctggcagacg cagagcctgt      840

agagccatga ggcctaagac cgccggaaga agaaacgacc tgcacgacga tagaaccgcc      900

agcgctcaca agaccagcag acagaagaga agggccgagt acgccagggt gcaagagctg      960

tacaagaagt gcagatccag agccgccgct gaagtgattg atggtgcttg tggtggcgtg     1020

ggccacagcc tggaagagat ggaaacctat tggcggccca tcctggaaag agtgtctgac     1080

gctcctggac caacacctga agctctgcat gctctgggca gagctgagtg gcatggcggc     1140

aatagagatt acacccagct gtggaagccc atcagcgtgg aagaaatcaa ggccagcaga     1200

ttcgactggc ggacaagccc tggacctgat ggcattagat ctggacagtg gcgggctgtg     1260

cctgtgcacc tgaaggccga aatgttcaac gcctggatgg ccagaggcga gatccctgag     1320

atcctgagac agtgcagaac cgtgttcgtg cccaaggtgg aaagacctgg cggaccaggc     1380

gagtacagac ccatctctat cgccagcatt cctctgcggc acttccactc tatcctggct     1440

cggagacttc tggcctgctg tcctcctgat gccagacaga gaggctttat ctgcgccgac     1500

ggcaccctgg aaaattctgc agtgctggat gccgtgctgg gcgactctcg gaagaaactg     1560

agagaatgtc acgtggccgt cctggacttc gccaaggcct ttgatacagt gtctcacgag     1620

gccctggtgg aactgctgag actgagggga atgcctgagc agttctgtgg ctatatcgcc     1680

cacctgtacg acaccgcctc taccacactg gccgtgaaca atgagatgag cagccccgtg     1740

aaagttggca gaggcgttag acagggcgac cctctgagcc ccatcctgtt caatgtggtc     1800

atggatctga tcctggccag cctgcctgag agagtgggct atagactgga aatggaactg     1860

gtgtctgccc tggcctacgc cgatgatctg gttctgcttg ccggcagcaa agtgggcatg     1920

caagagtcta tcagcgccgt ggattgcgtg ggcagacaga tgggcctgcg cctgaattgc     1980

agaaaaagcg ccgtgctgag catgatcccc gatggccaca gaaagaagca ccactacctg     2040

accgagcgga ccttcaatat cggcggcaag cctctgagac aggtgtcctg tgttgagaga     2100

tggcggtatc tgggcgtcga ctttgaggcc tctggctgtg tgacactgga acactctatc     2160

agcagcgccc tgaacaacat cagcagagcc cctctgaagc ctcagcagcg gctggaaatt     2220

ctgagagccc atctgatccc tcggttccag cacggatttg tgctgggcaa catctccgac     2280

gaccggctga gaatgctgga cgtgcagatc agaaaagccg tcggccagtg gctgagactt     2340

cctgcagatg tgcctaaggc ctactatcac gctgctgtgc aagatggcgg cctggctatt     2400

ccttctgtgc gcgccacaat tcccgacctg atcgtgcgaa gattcggcgg acttgatagc     2460

tctccttgga gcgtggccag agctgccgcc aagagcgata agatccggaa aaagctgcgc     2520

tgggcctgga agcagctgcg gagattttct agagtggaca gcaccacaca gcggcctagt     2580

gtgcggctgt tttggagaga acatctgcac gcctccgtgg acggcagaga gctgagagaa     2640

agcaccagaa cacccaccag caccaagtgg atcagagaga gatgcgccca gatcacaggc     2700

cgggatttcg tgcagttcgt gcacacccat atcaacgccc tgccatccag aatcaggggc     2760

agcagaggta gaagaggcgg aggcgaaagc agcctgacat gtagagccgg ctgtaaagtg     2820

cgcgagacaa cagcccacat cctgcagcag tgtcatagaa cacacggcgg cagaatcctg     2880

cggcacaaca agattgtgtc cttcgtggcc aaggccatgg aagagaacaa gtggaccgtg     2940

gaactggaac ccagactgag aacaagcgtg ggcctgagaa agcccgacat cattgcctct     3000

cgagatggcg tgggagtgat cgtggatgtg caggttgtgt caggccagag aagcctggac     3060

gagctgcaca gagagaagcg gaacaaatac ggcaaccacg gcgagctggt tgaactggtt     3120

gcaggcagac tgggactgcc aaaagccgag tgtgtgcggg ccacctcttg taccatttct     3180

tggagaggcg tgtggtccct gaccagctac aaagagctgc ggtccatcat cggactgaga     3240

gagcctacac tgcagatcgt ccccattctg gccctgagag gcagccacat gaattggacc     3300

cgcttcaacc agatgaccag cgtgatggga ggcggcgttg ga                        3342


<210>  19
<211>  2221
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 19 - pTwist vector backbone

<400>  19
aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca       60

tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat      120

ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa      180

agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg      240

cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga      300

ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg      360

tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg      420

gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc      480

gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg      540

gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca      600

ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt      660

ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag      720

ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg      780

gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc      840

ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc      900

agcgtaaatg ggtagggggc ttcaaatcgt cctcgtgata ccaattcgga gcctgctttt      960

ttgtacaaac ttgttgataa tggcaattca aggatcttca cctagatcct tttaaattaa     1020

aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa     1080

tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc     1140

tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct     1200

gcaatgatac cgcgagagcc acgctcaccg gctccagatt tatcagcaat aaaccagcca     1260

gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt     1320

aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt     1380

gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc     1440

ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc     1500

tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt     1560

atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact     1620

ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc     1680

ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt     1740

ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg     1800

atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct     1860

gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa     1920

tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt     1980

ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc     2040

acatttcccc gaaaagtgcc agatacctga aacaaaaccc atcgtacggc caaggaagtc     2100

tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg ttgtaaaacg     2160

acggccagtc atgcataatc cgcacgcatc tggaataagg aagtgccatt ccgcctgacc     2220

t                                                                     2221


<210>  20
<211>  222
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 20 - polI - RNA polymerase I promoter

<400>  20
gccgggaggg cgtccccggc ccggcgctgc tcccgcgtgt gtcctggggt tgaccagagg       60

gccccgggcg ctccgtgtgt ggctgcgatg gtggcgtttt tggggacagg tgtccgtgtc      120

gcgcgtcgcc tgggccggcg gcgtggtcgg tgacgcgacc tcccggcccc gggggaggta      180

tatctttcgc tccgagtcgg cattttgggc cgccgggtta tt                         222


<210>  21
<211>  265
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 21 - R2OI 5'UTR

<400>  21
cgcacagggg acacagagcc tgcccaagta ccgctcccga gggagcggga aacggggggg       60

tgactatccc ctggggtccg gcgagagcgc tggtctacgg accaggggtg gctgtgggca      120

ggctgctcct caggccagtt gattagttac gcatgggctg tacctccacg tggtcccgct      180

ggtaacgact tgtcggctaa atcagcccgc ccaccatctg ggatatggtt gaccgtctaa      240

ccccagtact caggtcacaa acaaa                                            265


<210>  22
<211>  3831
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 22 - R2OI for RNA R2OI ORF

<400>  22
atgggaacag atacagtgta tgtcggccag gactaccctt ctggcttatc aaaacgggta       60

ccagcacggt tagtggcggg accgatgctg cgagagcgaa gctgtcacgc ccatgtgttt      120

agggctggac acatgtggaa ctggcgaacc agccttccga gcgggcgctg ggaccagccc      180

gctttggaga agtctcgggt cctaacccgg tcggtggcga cggccaccga ccccgaaatt      240

acctcttacc caggaaagtc cgtatcgaca agtacgcagg ttcaggagga ggactggtgt      300

agccgggaga gcgggtggat ctcgccagga cttgctcctg aagaaccctc ggtggtgtcc      360

gaaattacag cctccatggt agcgacaatg agggtagcaa ccgaggaggt cgtgctggaa      420

ccacagcctg aacaggtcgt cacaatactg ccggagcatg gtcgaaacgt tcctccgggg      480

ctggcagaac aggacaccgc cagccccata gaagtctcgg tgctcctccc agacctcgct      540

gagaactgcc cattgtgtgg cgtgccgagc gggggcctac gcttgctcgg gaagcatttt      600

gctgtccgac atgcgggggt gcctgtaacg tatgagtgcc gtaagtgtgc gtggcggagc      660

cccaacagcc actcaatctc gtgtcacgtc cccaaatgcc gggggcgtgc gcggatgccc      720

agtggcgatc cagggatcgc ctgcgatctc tgtgaagccc ggtttgccac ggaggttggg      780

gtcgcccaac acaagcggca cgttcatccg gtggagtgga acaaggtgag gctggaaagg      840

agaggtgcgc gcggaggggg aattaaggcg acgaagctct ggagtgtagc ggaggtagag      900

acgctaatcc ggctcatccg tgagcacgga gattcaggtg ccacttacca gctcattgcc      960

gatgagctgg gaaggggcaa gacggccgaa caggtgagga gtaaaaagag gctcctgcgc     1020

atagatacgg caagcaatag cccagatgat gcagaggttg aggaggagag gttggaatct     1080

ctggcggttc ggtcctcgtc acggtcaccc ccgagcctgg tggcgaccag ggtcagggag     1140

gcagttgcca ggggtgaatc agaaggtggc gaggagatca gggctattgc tgctctcatt     1200

agggacgtag atcagaatcc ttgtctgatt gaaacctcgg cgtcggacat catctcgaag     1260

ctgggaagga gggtggatgg gcccaagaga cccaggcccg ttgtcagaga acagacccaa     1320

gagaagggat gggtaaggcg gcttgcccgg cggaaaaggg agtacagaga agcgcagtac     1380

ctgtactcaa gggatcaagc aaggctggcg gcccagatcc tcgatggtgc cgccagccag     1440

gaatgcgccc tcccggtgga ccaggtctac ggagcgttcc gtgagaaatg ggaaaccgta     1500

gggcagttcc acggacttgg tgagttccgg acgggtgcac gcgcagacaa ctgggagttc     1560

tactctccaa ttctggcggc tgaggtgaaa gaaaacctaa tgagaatggc taacggcacg     1620

gccccgggac cagacaggat aagcaaaaag gctctgcttg actgggaccc ccggggtgag     1680

caactggcac ggctgtacac gacgtggctg atcggtgggg tcataccaag ggtcttcaag     1740

gagtgcagga ctaagctgct accgaaatcc agcgacccgg tcgagttgca ggacatcggt     1800

ggatggaggc cggtgacgat tgggtcgatg gtgactaggc tgttcagtcg gattctaacg     1860

atgaggctaa cccgagcctg tccgatcaat ccgaggcagc gcggtttctt ggcctcctcg     1920

agtggatgcg cggaaaacct gttgatcttt gacgagatcg tcaggcgctc gaggcgggac     1980

ggggggccgc tggcagtggt gtttgtggac tttgcgaggg cctttgactc catctcacat     2040

gaacatatcc tgtgtgttct cgaagaaggc gggcttgaca ggcacgttat cgggttgatc     2100

cgaaactcgt acgtggattg cgtgaccagg gtgggttgtg tcgagggcat gacaccacca     2160

atacaaatga aggttggagt gaagcaggga gaccccatgt cccccttgct cttcaacctg     2220

gctatggatc ccctcatcca taaactcgag acggccggaa ctggactgaa atggggcgat     2280

ctttcaatcg ccacgctggc ctttgccgac gatctggtgc tggtgagtga ctctgaggaa     2340

ggcatgggga ggagtctcgg gattttggag aagttttgcc aactgactgg gctgagggtt     2400

cagcccagga agtgtcacgg tttctttatg gacaagggcg tggtgaacgg ctgtggaacc     2460

tgggaaatct gtgggtcacc gatccacatg attcccccgg gggaatcagt tcgttatttg     2520

ggagtccagg taggcccggg gcgcggcgtg atggaaccgg atcttatccc tacggtccac     2580

acgtggatcg aaaggatctc ggaggctcct ctaaagccct cacaacgcat gagggttttg     2640

aactcattcg ctctcccccg gataatttac caggccgatc tagggaaggt tacggtaacc     2700

aaattggccc agatagatgg gattgtccgg aaggctgtga agaagtggct ccatttgtca     2760

ccatccacgt gcaatggact gctgtattca cggaaccgcg acggtggttt gggcctccta     2820

aagctggaaa gactaatccc atccgtgcgc acgaagcgta tctatcggat gtccaggtct     2880

ccggatatct ggacacggcg aatgaccagc cattctgtgt caaaatctga ctgggagatg     2940

ttgtgggtcc aagcgggagg tgagaggggc agtgcacctg taatgggtgc cgtggaggct     3000

gccccgaccg atgtggagag atcgccagac tacccagact ggcggcgtga ggaaaacctg     3060

gcatggtcgg ccctgcgggt gcagggtgtg ggtgcagacc agtttcgagg cgacaggacc     3120

agcagctctt ggatcgccga gcccgcttcg gttgggttcg cgcagcgcca ctggttggct     3180

gccctggcgc tgagggctgg ggtgtatccg actcgggagt ttctggctcg gggtaaggaa     3240

aagtcaggag cagcttgcag acgctgcccg gccaggttgg aatcatgttc acacatactt     3300

gggcaatgtc cgttcgttca ggcgaacaga attgcgaggc acaacaaggt gtgtgtgctc     3360

ttggccacgg aggcggagag gttcggctgg acggtaataa gggagttccg tcttgaggac     3420

gccgctggcg gtctcaagat acccgacctg gtttgcaaga aggccgacac agttctcatt     3480

gtcgacgtga ccgtccggta cgagatggat ggagagacgc taaaaagggc cgcatcggag     3540

aaggtgaaac actatctccc agtagggcaa cagataacgg acaaggtcgg agggcgttgc     3600

tttaaagtca tggggttccc tgtaggtgct aggggaaagt ggccggcgag caacaacaca     3660

gttttggctg agttaggcgt ccctgcaggt cggatgagga cctttgccag gctggtgagc     3720

cggaggactc ttctttattc tttggatata ttgagggact tcatgcgtga gccggccggc     3780

aggggaactc gggttgctct catccctgcg gcaacgggtg ccgcgaattg a              3831


<210>  23
<211>  108
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 23 - R2OI 3'UTR

<400>  23
gggggacagc tgggagtctc ggcatgatta caaatcttgc gctgcactcg gatgtcgtcc       60

ccgtgacgga cacattaatc cggaaagcga gtggtgactc gcctcaag                   108


<210>  24
<211>  3345
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 24 - R2 for RNA ORF

<400>  24
atgatggcga gcaccgcact gtcccttatg ggacggtgta acccggatgg ctgtacacgt       60

ggtaaacacg tgacagcagc cccgatggac ggaccgcgag gaccgtcaag cctagcaggt      120

accttcgggt ggggccttgc gatacctgcg ggcgaaccct gtggtcgggt ttgcagcccg      180

gccacagtgg gtttttttcc tgttgcaaaa aagtcaaata aagaaaatag acctgaagcc      240

tctggcctcc cgctggagtc agagaggaca ggcgataacc cgactgtgcg gggttccgcc      300

ggcgcagatc ctgtgggtca ggatgcgcct ggttggacct gccagttctg cgaacgaacc      360

ttttcgacca acaggggttt gggtgtccac aagcgtagag cccaccctgt tgagaccaat      420

acggatgccg ctccgatgat ggtgaagcgg cggtggcatg gcgaggaaat cgacctcctc      480

gctcgcaccg aggccaggtt gctcgctgag cggggtcagt gctcgggtgg agacctcttt      540

ggcgcgcttc cagggtttgg aagaactctg gaagcgatta agggacaacg gcggagggag      600

ccttatcggg cattggtgca agcgcacctt gcccgatttg gttcccagcc gggtccctcg      660

tcgggggggt gctcggccga gcctgacttc cggcgggctt ctggagctga ggaagcgggc      720

gaggaacgat gcgccgaaga cgccgctgcc tatgatccat ccgcagtcgg tcagatgtcg      780

cccgatgccg ctcgggttct ctccgaactc cttgagggtg cggggagaag acgagcgtgc      840

agggctatga gacccaagac tgcagggcgg cgaaacgatt tgcacgatga tcggacagct      900

agtgcccaca aaaccagtag acaaaagcgc agggcagagt acgcgcgtgt gcaggaactg      960

tacaagaagt gtcgcagcag agcagcagct gaggtgatcg atggcgcgtg tgggggtgtc     1020

ggacactcgc tcgaggagat ggagacctat tggcgaccta tcctcgagag agtgtccgat     1080

gcacctgggc ctacaccgga agctcttcac gccctagggc gtgcggagtg gcacgggggc     1140

aatcgcgact acacccagct gtggaagccg atctcggtgg aagagatcaa ggcctcccgc     1200

tttgactggc gaacttcgcc gggcccggac ggtatacgtt cgggtcagtg gcgtgcggtt     1260

cctgtgcact tgaaggcgga aatgttcaat gcatggatgg cacgaggcga aatacccgaa     1320

attctacggc agtgccgaac cgtctttgta cctaaggtgg agagaccagg tggaccgggg     1380

gaatatcgac cgatctcgat cgcgtcgatt cccctgagac actttcactc catcttggcc     1440

cggaggctgt tggcttgctg cccccctgat gcacgacagc gcggatttat ctgcgccgac     1500

ggtacgctgg agaattccgc agtactggac gcggtgcttg gggatagcag gaagaagctg     1560

cgggaatgtc acgtggcggt gctagacttc gccaaggcat ttgacacagt gtctcacgag     1620

gcacttgtcg aattgctgag gttgaggggc atgcccgaac agttctgcgg ctacattgct     1680

cacctatacg atacggcgtc caccacctta gccgtgaaca atgaaatgag cagccctgta     1740

aaagtgggac gaggggttcg tcaaggggac cctctgtcgc cgatactctt caacgtggtg     1800

atggacctca tcctggcttc cctgccggag agggtcgggt ataggttgga gatggaactc     1860

gtgtccgctc tggcctatgc tgacgaccta gtcctgcttg cggggtcgaa ggtagggatg     1920

caggagtcca tctctgctgt ggactgtgtc ggtaggcaga tgggcctacg cctgaattgc     1980

aggaaaagcg cggttctgtc tatgataccg gatggccacc gcaagaagca tcactacctg     2040

actgagcgaa ccttcaatat tggaggtaag ccgctcaggc aggtgagttg tgttgagcgg     2100

tggcgatatc ttggtgtcga ttttgaggcc tctggatgcg tgacattaga gcatagtatc     2160

agtagtgctc tgaataacat ctcaagggca cctctcaaac cccaacagag gttggagatt     2220

ttgagagctc atctgattcc gagattccag cacggttttg tgcttggaaa catctcggat     2280

gaccgattga gaatgctcga tgtccaaatc cggaaagcag tcggacagtg gctaaggcta     2340

ccggcggatg tgcccaaggc atattatcac gccgcagttc aggacggcgg cttagcgatc     2400

ccatcggtgc gagcgaccat cccggacctc attgtgaggc gtttcggggg gctcgactcg     2460

tcaccatggt cagtggcaag agccgccgcc aaatctgata agattcgtaa gaaactgcgg     2520

tgggcctgga aacagctccg caggttcagc cgtgttgact ccacaacgca acgaccatct     2580

gtgcgcttgt tttggcgaga acatctgcat gcatctgttg atggacgcga acttcgcgaa     2640

tccacacgca ccccgacatc cacaaagtgg attagggagc gatgcgcgca gataaccgga     2700

cgggacttcg tgcagttcgt gcacactcat atcaacgccc tcccatcccg cattcgcgga     2760

tcgagagggc gtagaggtgg gggtgagtct tcgttgacct gccgtgctgg ttgcaaggtt     2820

agggagacga cggctcacat cctacaacag tgtcacagaa cacacggcgg ccggattcta     2880

cgacacaaca agattgtatc tttcgtggcg aaagccatgg aagagaacaa gtggacggtt     2940

gagctggagc cgaggctacg aacatcggtt ggtctccgta agccggatat tatcgcctcc     3000

agggatggtg tcggagtgat cgtggacgtg caggtggtct cgggccagcg atcgcttgac     3060

gagctccacc gtgagaaacg taataaatac gggaatcacg gggagctggt tgagttggtc     3120

gcaggtagac taggacttcc gaaagctgag tgcgtgcgag ccacttcgtg cacgatatct     3180

tggaggggag tatggagcct gacttcttat aaggagttaa ggtccataat cgggcttcgg     3240

gaaccgacac tacaaatcgt tccgatactg gcgttgagag gttcacacat gaactggacc     3300

aggttcaatc agatgacgtc cgtcatgggg ggcggcgttg gttga                     3345


<210>  25
<211>  620
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 25 - R2 5'UTR

<400>  25
gcgggagtaa ctatgactct cttaggggcg atacgcataa ttttaatttt tcgattcaaa       60

tccagtcgtc ttaatctggt gaccagtggc gcggtcacca gtatagtgca caggacgtga      120

atggctccga ggctggcgga gtcactcact ataagtgtga gagacgatgt cctgtgccaa      180

gtatacgtcc aaccctaacg ggttaagtga aattagttgc tcataacagg gacggtgtac      240

ctgtttgctc gtggctggct atcgaatgga cgggaccaat acacccccct gttagtaatg      300

gggtaagaga gagcggtctg aaactatggc cgagatcacg acgccccact cctacccata      360

acctgcacgt ggtaccgccg cacattgacc gatacgggag gaggggcagc acttgaatca      420

cgtagtcttg gtgtagccat tgcgggacta cagccctcgt aagtgccgcc ttagaacgca      480

acggggcaat aggtgggccg gggcgctagc gggggggagt aatctcccct gttggcgtgc      540

accgcactgc tccctctggg ggcagtgtca tccggaaaca ggtgggccgg ggcgccacca      600

ggggggagca atccctcctg                                                  620


<210>  26
<211>  248
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 26 - R2 3'UTR

<400>  26
gccttgcaca gtagtccagc ggtaagggtg tagatcaggc ccgtctgttt ctcccccgga       60

gctcgctccc ttggcttccc ttatatattt taacatcaga aacagacatt aaacatctac      120

tgatccaatt tcgccggcgt acggccacga tcgggagggt gggaatctcg ggggtcttcc      180

gatcctaatc catgatgatt acgacctgag tcactaaaga cgatggcatg atgatccggc      240

gatgaaaa                                                               248


<210>  27
<211>  106
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 27 - r106 homology arm - 106 bp sequence 28S human 
       ribosomal gene upstream from the insertion site

<400>  27
gcgggtgttg acgcgatgtg atttctgccc agtgctctga atgtcaaagt gaagaaattc       60

aatgaagcgc gggtaaacgg cgggagtaac tatgactctc ttaagg                     106


<210>  28
<211>  100
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 28 - r100 homology arm - 100bp sequence of 28S human 
       ribosomal gene downstream from the insertion site

<400>  28
tagccaaatg cctcgtcatc taattagtga cgcgcatgaa tggatgaacg agattcccac       60

tgtccctacc tactatccag cgaaaccaca gccaagggaa                            100


<210>  29
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 29 - r30 homology arm - 30bp sequence of 28S human 
       ribosomal gene downstream from the insertion site

<400>  29
tagccaaatg cctcgtcatc taattagtga                                        30


<210>  30
<211>  15
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 30 - r15 homology arm - 15bp sequence of 28S human 
       ribosomal gene downstream from the insertion site

<400>  30
tagccaaatg cctcg                                                        15


<210>  31
<211>  10
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 31 - r10 homology arm - 10bp sequence of 28S human 
       ribosomal gene downstream from the insertion site

<400>  31
tagccaaatg                                                              10


<210>  32
<211>  35
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 32 - polI terminator - 35bp, RNA polymerase I 
       terminator

<400>  32
tcccccccaa cttcggaggt cgaccagtac tccgg                                  35


<210>  33
<211>  58
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 33 - NGS Forward Primer


<220>
<221>  misc_feature
<222>  (34)..(36)
<223>  n is a, c, g, or t

<400>  33
tcgtcggcag cgtcagatgt gtataagaga cagnnngtag cctcagtctt cccatcag         58


<210>  34
<211>  55
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 34 - NGS Reverse Primer


<220>
<221>  misc_feature
<222>  (35)..(37)
<223>  n is a, c, g, or t

<400>  34
gtctcgtggg ctcggagatg tgtataagag acagnnncag cagcaagcag cactc            55


<210>  35
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 35 - EMX guide RNA sequence

<400>  35
gagtccgagc agaagaagaa                                                   20


<210>  36
<211>  123
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 36 - HA-NLS-XTEN sequence

<400>  36
ggatcctacc catacgatgt tccagattac gcggccgctc caaaaaagaa aagaaaagtt       60

gaattcggcg gcagcagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa      120

agt                                                                    123


<210>  37
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 37 - XTEN linker sequence

<400>  37
agcggcagcg agactcccgg gacctcagag tccgccacac ccgaaagt                    48


<210>  38
<211>  171
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 38 - HA-NLS-32aa sequence

<400>  38
ggatcctacc catacgatgt tccagattac gcggccgctc caaaaaagaa aagaaaagtt       60

gaattcggcg gcagctctgg tggttcttct ggtggttcta gcggcagcga gactcccggg      120

acctcagagt ccgccacacc cgaaagttct ggtggttctt ctggtggttc t               171


<210>  39
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 39 - 32aa linker sequence

<400>  39
tctggtggtt cttctggtgg ttctagcggc agcgagactc ccgggacctc agagtccgcc       60

acacccgaaa gttctggtgg ttcttctggt ggttct                                 96


<210>  40
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 40 - SpCas9 Human codon optimized

<400>  40
atggacaaga agtatagcat cggcctggat atcggcacaa actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccaagcaag aagtttaagg tgctgggcaa caccgataga      120

cactccatca agaagaatct gatcggcgcc ctgctgttcg actctggcga gacagccgag      180

gccacacggc tgaagagaac cgcccggaga aggtatacac gccggaagaa taggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgattcttt ctttcaccgc      300

ctggaggaga gcttcctggt ggaggaggat aagaagcacg agcggcaccc tatctttggc      360

aacatcgtgg acgaggtggc ctatcacgag aagtacccaa caatctatca cctgaggaag      420

aagctggtgg actccaccga taaggccgac ctgcgcctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca ctttctgatc gagggcgatc tgaacccaga caatagcgat      540

gtggacaagc tgttcatcca gctggtgcag acctacaatc agctgtttga ggagaacccc      600

atcaatgcct ctggagtgga cgcaaaggca atcctgagcg ccagactgtc caagtctaga      660

aggctggaga acctgatcgc ccagctgcca ggcgagaaga agaacggcct gtttggcaat      720

ctgatcgccc tgtccctggg cctgacaccc aacttcaagt ctaattttga tctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tatgacgatg acctggataa cctgctggcc      840

cagatcggcg atcagtacgc cgacctgttc ctggccgcca agaatctgtc tgacgccatc      900

ctgctgagcg atatcctgcg cgtgaacacc gagatcacaa aggcccccct gagcgcctcc      960

atgatcaaga gatatgacga gcaccaccag gatctgaccc tgctgaaggc cctggtgagg     1020

cagcagctgc ctgagaagta caaggagatc ttctttgatc agagcaagaa tggatacgca     1080

ggatatatcg acggaggagc atcccaggag gagttctaca agtttatcaa gcctatcctg     1140

gagaagatgg acggcacaga ggagctgctg gtgaagctga atcgggagga cctgctgagg     1200

aagcagcgca cctttgataa cggcagcatc cctcaccaga tccacctggg agagctgcac     1260

gcaatcctgc gccggcagga ggacttctac ccatttctga aggataaccg ggagaagatc     1320

gagaagatcc tgacattcag aatcccctac tatgtgggac ctctggcccg gggcaatagc     1380

agatttgcct ggatgacccg caagtccgag gagacaatca caccctggaa cttcgaggag     1440

gtggtggata agggcgcctc tgcccagagc ttcatcgagc ggatgaccaa ttttgacaag     1500

aacctgccta atgagaaggt gctgccaaag cactctctgc tgtacgagta tttcaccgtg     1560

tataacgagc tgacaaaggt gaagtacgtg accgagggca tgagaaagcc tgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttta agaccaatag gaaggtgaca     1680

gtgaagcagc tgaaggagga ctatttcaag aagatcgagt gttttgattc tgtggagatc     1740

agcggcgtgg aggacaggtt taacgcctcc ctgggcacct accacgatct gctgaagatc     1800

atcaaggata aggacttcct ggacaacgag gagaatgagg atatcctgga ggacatcgtg     1860

ctgaccctga cactgtttga ggatagggag atgatcgagg agcgcctgaa gacatatgcc     1920

cacctgttcg atgacaaagt gatgaagcag ctgaagagaa ggcgctacac cggatggggc     1980

cggctgagca gaaagctgat caatggcatc cgcgacaagc agtctggcaa gacaatcctg     2040

gactttctga agagcgatgg cttcgccaac cggaacttca tgcagctgat ccacgatgac     2100

tccctgacct tcaaggagga tatccagaag gcacaggtgt ctggacaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggctct cctgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaagtg atgggcaggc acaagccaga gaacatcgtg     2280

atcgagatgg cccgcgagaa tcagaccaca cagaagggcc agaagaactc ccgggagaga     2340

atgaagagaa tcgaggaggg catcaaggag ctgggctctc agatcctgaa ggagcacccc     2400

gtggagaaca cacagctgca gaatgagaag ctgtatctgt actatctgca gaatggccgg     2460

gatatgtacg tggaccagga gctggatatc aacagactgt ctgattatga cgtggatcac     2520

atcgtgccac agtccttcct gaaggatgac tctatcgaca ataaggtgct gaccaggagc     2580

gacaagaacc gcggcaagtc cgataatgtg ccctctgagg aggtggtgaa gaagatgaag     2640

aactactgga ggcagctgct gaatgccaag ctgatcacac agaggaagtt tgataacctg     2700

accaaggcag agaggggagg actgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga caagacagat cacaaagcac gtggcccaga tcctggattc tagaatgaac     2820

acaaagtacg atgagaatga caagctgatc agggaggtga aagtgatcac cctgaagtcc     2880

aagctggtgt ctgactttag gaaggatttc cagttttata aggtgcgcga gatcaacaat     2940

tatcaccacg cccacgacgc ctacctgaac gccgtggtgg gcacagccct gatcaagaag     3000

taccctaagc tggagtccga gttcgtgtac ggcgactata aggtgtacga tgtgcgcaag     3060

atgatcgcca agtctgagca ggagatcggc aaggccaccg ccaagtattt cttttacagc     3120

aacatcatga atttctttaa gaccgagatc acactggcca atggcgagat caggaagcgc     3180

ccactgatcg agacaaacgg cgagacaggc gagatcgtgt gggacaaggg cagggatttt     3240

gccaccgtgc gcaaggtgct gagcatgccc caagtgaata tcgtgaagaa gaccgaggtg     3300

cagacaggcg gcttctccaa ggagtctatc ctgcctaagc ggaactccga taagctgatc     3360

gccagaaaga aggactggga ccccaagaag tatggcggct tcgacagccc tacagtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cgcagctcct tcgagaagaa tcctatcgac     3540

tttctggagg ccaagggcta taaggaggtg aagaaggacc tgatcatcaa gctgccaaag     3600

tactctctgt ttgagctgga gaacggaagg aagagaatgc tggcaagcgc cggagagctg     3660

cagaagggca atgagctggc cctgccctcc aagtacgtga acttcctgta tctggcctcc     3720

cactacgaga agctgaaggg ctctcctgag gataacgagc agaagcagct gtttgtggag     3780

cagcacaagc actatctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgccaatct ggataaggtg ctgtccgcct acaacaagca ccgggataag     3900

ccaatcagag agcaggccga gaatatcatc cacctgttta ccctgacaaa cctgggagca     3960

ccagcagcct tcaagtattt tgacaccaca atcgacagga agcggtacac cagcacaaag     4020

gaggtgctgg acgccacact gatccaccag tccatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgattga                                         4107


<210>  41
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SEQ ID NO: 41 - Dead_SpCas9 D10A and N863A mutations human codon 
       optimized

<400>  41
atggacaaga agtatagcat cggcctggcc atcggcacaa actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccaagcaag aagtttaagg tgctgggcaa caccgataga      120

cactccatca agaagaatct gatcggcgcc ctgctgttcg actctggcga gacagccgag      180

gccacacggc tgaagagaac cgcccggaga aggtatacac gccggaagaa taggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgattcttt ctttcaccgc      300

ctggaggaga gcttcctggt ggaggaggat aagaagcacg agcggcaccc tatctttggc      360

aacatcgtgg acgaggtggc ctatcacgag aagtacccaa caatctatca cctgaggaag      420

aagctggtgg actccaccga taaggccgac ctgcgcctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca ctttctgatc gagggcgatc tgaacccaga caatagcgat      540

gtggacaagc tgttcatcca gctggtgcag acctacaatc agctgtttga ggagaacccc      600

atcaatgcct ctggagtgga cgcaaaggca atcctgagcg ccagactgtc caagtctaga      660

aggctggaga acctgatcgc ccagctgcca ggcgagaaga agaacggcct gtttggcaat      720

ctgatcgccc tgtccctggg cctgacaccc aacttcaagt ctaattttga tctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tatgacgatg acctggataa cctgctggcc      840

cagatcggcg atcagtacgc cgacctgttc ctggccgcca agaatctgtc tgacgccatc      900

ctgctgagcg atatcctgcg cgtgaacacc gagatcacaa aggcccccct gagcgcctcc      960

atgatcaaga gatatgacga gcaccaccag gatctgaccc tgctgaaggc cctggtgagg     1020

cagcagctgc ctgagaagta caaggagatc ttctttgatc agagcaagaa tggatacgca     1080

ggatatatcg acggaggagc atcccaggag gagttctaca agtttatcaa gcctatcctg     1140

gagaagatgg acggcacaga ggagctgctg gtgaagctga atcgggagga cctgctgagg     1200

aagcagcgca cctttgataa cggcagcatc cctcaccaga tccacctggg agagctgcac     1260

gcaatcctgc gccggcagga ggacttctac ccatttctga aggataaccg ggagaagatc     1320

gagaagatcc tgacattcag aatcccctac tatgtgggac ctctggcccg gggcaatagc     1380

agatttgcct ggatgacccg caagtccgag gagacaatca caccctggaa cttcgaggag     1440

gtggtggata agggcgcctc tgcccagagc ttcatcgagc ggatgaccaa ttttgacaag     1500

aacctgccta atgagaaggt gctgccaaag cactctctgc tgtacgagta tttcaccgtg     1560

tataacgagc tgacaaaggt gaagtacgtg accgagggca tgagaaagcc tgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttta agaccaatag gaaggtgaca     1680

gtgaagcagc tgaaggagga ctatttcaag aagatcgagt gttttgattc tgtggagatc     1740

agcggcgtgg aggacaggtt taacgcctcc ctgggcacct accacgatct gctgaagatc     1800

atcaaggata aggacttcct ggacaacgag gagaatgagg atatcctgga ggacatcgtg     1860

ctgaccctga cactgtttga ggatagggag atgatcgagg agcgcctgaa gacatatgcc     1920

cacctgttcg atgacaaagt gatgaagcag ctgaagagaa ggcgctacac cggatggggc     1980

cggctgagca gaaagctgat caatggcatc cgcgacaagc agtctggcaa gacaatcctg     2040

gactttctga agagcgatgg cttcgccaac cggaacttca tgcagctgat ccacgatgac     2100

tccctgacct tcaaggagga tatccagaag gcacaggtgt ctggacaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggctct cctgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaagtg atgggcaggc acaagccaga gaacatcgtg     2280

atcgagatgg cccgcgagaa tcagaccaca cagaagggcc agaagaactc ccgggagaga     2340

atgaagagaa tcgaggaggg catcaaggag ctgggctctc agatcctgaa ggagcacccc     2400

gtggagaaca cacagctgca gaatgagaag ctgtatctgt actatctgca gaatggccgg     2460

gatatgtacg tggaccagga gctggatatc aacagactgt ctgattatga cgtggatcac     2520

atcgtgccac agtccttcct gaaggatgac tctatcgaca ataaggtgct gaccaggagc     2580

gacaaggccc gcggcaagtc cgataatgtg ccctctgagg aggtggtgaa gaagatgaag     2640

aactactgga ggcagctgct gaatgccaag ctgatcacac agaggaagtt tgataacctg     2700

accaaggcag agaggggagg actgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga caagacagat cacaaagcac gtggcccaga tcctggattc tagaatgaac     2820

acaaagtacg atgagaatga caagctgatc agggaggtga aagtgatcac cctgaagtcc     2880

aagctggtgt ctgactttag gaaggatttc cagttttata aggtgcgcga gatcaacaat     2940

tatcaccacg cccacgacgc ctacctgaac gccgtggtgg gcacagccct gatcaagaag     3000

taccctaagc tggagtccga gttcgtgtac ggcgactata aggtgtacga tgtgcgcaag     3060

atgatcgcca agtctgagca ggagatcggc aaggccaccg ccaagtattt cttttacagc     3120

aacatcatga atttctttaa gaccgagatc acactggcca atggcgagat caggaagcgc     3180

ccactgatcg agacaaacgg cgagacaggc gagatcgtgt gggacaaggg cagggatttt     3240

gccaccgtgc gcaaggtgct gagcatgccc caagtgaata tcgtgaagaa gaccgaggtg     3300

cagacaggcg gcttctccaa ggagtctatc ctgcctaagc ggaactccga taagctgatc     3360

gccagaaaga aggactggga ccccaagaag tatggcggct tcgacagccc tacagtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cgcagctcct tcgagaagaa tcctatcgac     3540

tttctggagg ccaagggcta taaggaggtg aagaaggacc tgatcatcaa gctgccaaag     3600

tactctctgt ttgagctgga gaacggaagg aagagaatgc tggcaagcgc cggagagctg     3660

cagaagggca atgagctggc cctgccctcc aagtacgtga acttcctgta tctggcctcc     3720

cactacgaga agctgaaggg ctctcctgag gataacgagc agaagcagct gtttgtggag     3780

cagcacaagc actatctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgccaatct ggataaggtg ctgtccgcct acaacaagca ccgggataag     3900

ccaatcagag agcaggccga gaatatcatc cacctgttta ccctgacaaa cctgggagca     3960

ccagcagcct tcaagtattt tgacaccaca atcgacagga agcggtacac cagcacaaag     4020

gaggtgctgg acgccacact gatccaccag tccatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgattga                                         4107


<210>  42
<211>  401
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  R2_5'RNA - 50bp 5'UTR and 117aa of R2 ORF

<400>  42
tccggaaaca ggtgggccgg ggcgccacca ggggggagca atccctcctg atgatggcga       60

gcaccgcact gtcccttatg ggacggtgta acccggatgg ctgtacacgt ggtaaacacg      120

tgacagcagc cccgatggac ggaccgcgag gaccgtcaag cctagcaggt accttcgggt      180

ggggccttgc gatacctgcg ggcgaaccct gtggtcgggt ttgcagcccg gccacagtgg      240

gtttttttcc tgttgcaaaa aagtcaaata aagaaaatag acctgaagcc tctggcctcc      300

cgctggagtc agagaggaca ggcgataacc cgactgtgcg gggttccgcc ggcgcagatc      360

ctgtgggtca ggatgcgcct ggttggacct gccagttctg c                          401


<210>  43
<211>  679
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  R2OI_5'R2OI RNA - sequence required for protein binding composed 
       of 5'UTR and first 138aa of the R2OI ORF

<400>  43
cgcacagggg acacagagcc tgcccaagta ccgctcccga gggagcggga aacggggggg       60

tgactatccc ctggggtccg gcgagagcgc tggtctacgg accaggggtg gctgtgggca      120

ggctgctcct caggccagtt gattagttac gcatgggctg tacctccacg tggtcccgct      180

ggtaacgact tgtcggctaa atcagcccgc ccaccatctg ggatatggtt gaccgtctaa      240

ccccagtact caggtcacaa acaaaatggg aacagataca gtgtatgtcg gccaggacta      300

cccttctggc ttatcaaaac gggtaccagc acggttagtg gcgggaccga tgctgcgaga      360

gcgaagctgt cacgcccatg tgtttagggc tggacacatg tggaactggc gaaccagcct      420

tccgagcggg cgctgggacc agcccgcttt ggagaagtct cgggtcctaa cccggtcggt      480

ggcgacggcc accgaccccg aaattacctc ttacccagga aagtccgtat cgacaagtac      540

gcaggttcag gaggaggact ggtgtagccg ggagagcggg tggatctcgc caggacttgc      600

tcctgaagaa ccctcggtgg tgtccgaaat tacagcctcc atggtagcga caatgagggt      660

agcaaccgag gaggtcgtg                                                   679


<210>  44
<211>  1381
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Insert sequence - hPGK-eGFP-SV40 poly(A) signal

<400>  44
gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt       60

ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc      120

agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg      180

cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc      240

gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga      300

ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg      360

gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg      420

cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac      480

cgaatcaccg acctctctcc ccaggcaagt ttgtacaaaa aagcaggctg ccaccatggt      540

gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga      600

cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa      660

gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt      720

gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca      780

cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa      840

ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa      900

ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct      960

ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat     1020

caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca     1080

ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct     1140

gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct     1200

ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaaaaca     1260

acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa     1320

ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt     1380

a                                                                     1381


<210>  45
<211>  2258
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of spScaffold, 106bp homology arm, R2_5'RNA, hPGK-GFP-SV40
       polyA, R2_3'RNA, 30bp - 3' homology arm

<400>  45
tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact       60

tgaaaaagtg gcaccgagtc ggtgcttttt ttgcgggtgt tgacgcgatg tgatttctgc      120

ccagtgctct gaatgtcaaa gtgaagaaat tcaatgaagc gcgggtaaac ggcgggagta      180

actatgactc tcttaaggtc cggaaacagg tgggccgggg cgccaccagg ggggagcaat      240

ccctcctgat gatggcgagc accgcactgt cccttatggg acggtgtaac ccggatggct      300

gtacacgtgg taaacacgtg acagcagccc cgatggacgg accgcgagga ccgtcaagcc      360

tagcaggtac cttcgggtgg ggccttgcga tacctgcggg cgaaccctgt ggtcgggttt      420

gcagcccggc cacagtgggt ttttttcctg ttgcaaaaaa gtcaaataaa gaaaatagac      480

ctgaagcctc tggcctcccg ctggagtcag agaggacagg cgataacccg actgtgcggg      540

gttccgccgg cgcagatcct gtgggtcagg atgcgcctgg ttggacctgc cagttctgcg      600

ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg      660

gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca      720

gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc      780

ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg      840

cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac      900

cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg      960

aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc     1020

ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc     1080

gaatcaccga cctctctccc caggcaagtt tgtacaaaaa agcaggctgc caccatggtg     1140

agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac     1200

gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag     1260

ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg     1320

accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac     1380

gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag     1440

gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac     1500

cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg     1560

gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc     1620

aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac     1680

taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg     1740

agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg     1800

gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaaacaa     1860

cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa     1920

taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta     1980

gccttgcaca gtagtccagc ggtaagggtg tagatcaggc ccgtctgttt ctcccccgga     2040

gctcgctccc ttggcttccc ttatatattt taacatcaga aacagacatt aaacatctac     2100

tgatccaatt tcgccggcgt acggccacga tcgggagggt gggaatctcg ggggtcttcc     2160

gatcctaatc catgatgatt acgacctgag tcactaaaga cgatggcatg atgatccggc     2220

gatgaaaata gccaaatgcc tcgtcatcta attagtga                             2258


<210>  46
<211>  2396
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of spScaffold, 106bp homology arm, R2OI_5'RNA, 
       hPGK-GFP-SV40 polyA, R2OI_3'RNA, 30bp - 3' homology arm

<400>  46
tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact       60

tgaaaaagtg gcaccgagtc ggtgcttttt ttgcgggtgt tgacgcgatg tgatttctgc      120

ccagtgctct gaatgtcaaa gtgaagaaat tcaatgaagc gcgggtaaac ggcgggagta      180

actatgactc tcttaaggcg cacaggggac acagagcctg cccaagtacc gctcccgagg      240

gagcgggaaa cgggggggtg actatcccct ggggtccggc gagagcgctg gtctacggac      300

caggggtggc tgtgggcagg ctgctcctca ggccagttga ttagttacgc atgggctgta      360

cctccacgtg gtcccgctgg taacgacttg tcggctaaat cagcccgccc accatctggg      420

atatggttga ccgtctaacc ccagtactca ggtcacaaac aaaatgggaa cagatacagt      480

gtatgtcggc caggactacc cttctggctt atcaaaacgg gtaccagcac ggttagtggc      540

gggaccgatg ctgcgagagc gaagctgtca cgcccatgtg tttagggctg gacacatgtg      600

gaactggcga accagccttc cgagcgggcg ctgggaccag cccgctttgg agaagtctcg      660

ggtcctaacc cggtcggtgg cgacggccac cgaccccgaa attacctctt acccaggaaa      720

gtccgtatcg acaagtacgc aggttcagga ggaggactgg tgtagccggg agagcgggtg      780

gatctcgcca ggacttgctc ctgaagaacc ctcggtggtg tccgaaatta cagcctccat      840

ggtagcgaca atgagggtag caaccgagga ggtcgtgggg ttgcgccttt tccaaggcag      900

ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc      960

cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc     1020

tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt     1080

gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc     1140

agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag     1200

cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg     1260

gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc     1320

tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca     1380

ggcaagtttg tacaaaaaag caggctgcca ccatggtgag caagggcgag gagctgttca     1440

ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg     1500

tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca     1560

ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc     1620

agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc     1680

ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc     1740

gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg     1800

acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca     1860

acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc     1920

acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg     1980

gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca     2040

aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga     2100

tcactctcgg catggacgag ctgtacaagt aaaaacaact tgtttattgc agcttataat     2160

ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat     2220

tctagttgtg gtttgtccaa actcatcaat gtatcttagg gggacagctg ggagtctcgg     2280

catgattaca aatcttgcgc tgcactcgga tgtcgtcccc gtgacggaca cattaatccg     2340

gaaagcgagt ggtgactcgc ctcaagtagc caaatgcctc gtcatctaat tagtga         2396


<210>  47
<211>  2279
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of SPACER, spScaffold, 106bp homology arm, R2_5'RNA, 
       hPGK-GFP-SV40 polyA, R2_3'RNA, 30bp - 3' homology arm

<400>  47
taattagtga cgcgcatgaa gtttaagagc tatgctggaa acagcatagc aagtttaaat       60

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttgcgggtg      120

ttgacgcgat gtgatttctg cccagtgctc tgaatgtcaa agtgaagaaa ttcaatgaag      180

cgcgggtaaa cggcgggagt aactatgact ctcttaaggt ccggaaacag gtgggccggg      240

gcgccaccag gggggagcaa tccctcctga tgatggcgag caccgcactg tcccttatgg      300

gacggtgtaa cccggatggc tgtacacgtg gtaaacacgt gacagcagcc ccgatggacg      360

gaccgcgagg accgtcaagc ctagcaggta ccttcgggtg gggccttgcg atacctgcgg      420

gcgaaccctg tggtcgggtt tgcagcccgg ccacagtggg tttttttcct gttgcaaaaa      480

agtcaaataa agaaaataga cctgaagcct ctggcctccc gctggagtca gagaggacag      540

gcgataaccc gactgtgcgg ggttccgccg gcgcagatcc tgtgggtcag gatgcgcctg      600

gttggacctg ccagttctgc gggttgcgcc ttttccaagg cagccctggg tttgcgcagg      660

gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca      720

cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc      780

cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc      840

ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg      900

gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc      960

gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt     1020

gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg     1080

cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccaggcaagt ttgtacaaaa     1140

aagcaggctg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc     1200

ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag     1260

ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc     1320

gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac     1380

cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag     1440

gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc     1500

gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc     1560

aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc     1620

gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc     1680

agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg     1740

ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag     1800

cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac     1860

gagctgtaca agtaaaaaca acttgtttat tgcagcttat aatggttaca aataaagcaa     1920

tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc     1980

caaactcatc aatgtatctt agccttgcac agtagtccag cggtaagggt gtagatcagg     2040

cccgtctgtt tctcccccgg agctcgctcc cttggcttcc cttatatatt ttaacatcag     2100

aaacagacat taaacatcta ctgatccaat ttcgccggcg tacggccacg atcgggaggg     2160

tgggaatctc gggggtcttc cgatcctaat ccatgatgat tacgacctga gtcactaaag     2220

acgatggcat gatgatccgg cgatgaaaat agccaaatgc ctcgtcatct aattagtga      2279


<210>  48
<211>  2417
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of SPACER, spScaffold, 106bp homology arm, R2OI_5'RNA, 
       hPGK-GFP-SV40 polyA, R2OI_3'RNA, 30bp - 3' homology arm

<400>  48
taattagtga cgcgcatgaa gtttaagagc tatgctggaa acagcatagc aagtttaaat       60

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttgcgggtg      120

ttgacgcgat gtgatttctg cccagtgctc tgaatgtcaa agtgaagaaa ttcaatgaag      180

cgcgggtaaa cggcgggagt aactatgact ctcttaaggc gcacagggga cacagagcct      240

gcccaagtac cgctcccgag ggagcgggaa acgggggggt gactatcccc tggggtccgg      300

cgagagcgct ggtctacgga ccaggggtgg ctgtgggcag gctgctcctc aggccagttg      360

attagttacg catgggctgt acctccacgt ggtcccgctg gtaacgactt gtcggctaaa      420

tcagcccgcc caccatctgg gatatggttg accgtctaac cccagtactc aggtcacaaa      480

caaaatggga acagatacag tgtatgtcgg ccaggactac ccttctggct tatcaaaacg      540

ggtaccagca cggttagtgg cgggaccgat gctgcgagag cgaagctgtc acgcccatgt      600

gtttagggct ggacacatgt ggaactggcg aaccagcctt ccgagcgggc gctgggacca      660

gcccgctttg gagaagtctc gggtcctaac ccggtcggtg gcgacggcca ccgaccccga      720

aattacctct tacccaggaa agtccgtatc gacaagtacg caggttcagg aggaggactg      780

gtgtagccgg gagagcgggt ggatctcgcc aggacttgct cctgaagaac cctcggtggt      840

gtccgaaatt acagcctcca tggtagcgac aatgagggta gcaaccgagg aggtcgtggg      900

gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg      960

ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag     1020

cgtcacccgg atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc     1080

cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc     1140

acgtctcact agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc     1200

gcgatgggct gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga     1260

aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg     1320

gtgttccgca ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg     1380

aatcaccgac ctctctcccc aggcaagttt gtacaaaaaa gcaggctgcc accatggtga     1440

gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg     1500

taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc     1560

tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga     1620

ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg     1680

acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg     1740

acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc     1800

gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg     1860

agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca     1920

aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact     1980

accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga     2040

gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg     2100

agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaaaacaac     2160

ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat     2220

aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttag     2280

ggggacagct gggagtctcg gcatgattac aaatcttgcg ctgcactcgg atgtcgtccc     2340

cgtgacggac acattaatcc ggaaagcgag tggtgactcg cctcaagtag ccaaatgcct     2400

cgtcatctaa ttagtga                                                    2417


<210>  49
<211>  2278
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of OMNI50_ TracrRNA, 106bp homology arm, R2_5'RNA, 
       hPGK-GFP-SV40 polyA, R2_3'RNA, 30bp - 3' homology arm

<400>  49
gtttgagagt tatgtaagaa attacatgac gagttcaaat aaaaatttat tcaaaccgcc       60

tatttatagg ccgcagatgt tctgcattat gcttgctatt gcaagctttt ttgcgggtgt      120

tgacgcgatg tgatttctgc ccagtgctct gaatgtcaaa gtgaagaaat tcaatgaagc      180

gcgggtaaac ggcgggagta actatgactc tcttaaggtc cggaaacagg tgggccgggg      240

cgccaccagg ggggagcaat ccctcctgat gatggcgagc accgcactgt cccttatggg      300

acggtgtaac ccggatggct gtacacgtgg taaacacgtg acagcagccc cgatggacgg      360

accgcgagga ccgtcaagcc tagcaggtac cttcgggtgg ggccttgcga tacctgcggg      420

cgaaccctgt ggtcgggttt gcagcccggc cacagtgggt ttttttcctg ttgcaaaaaa      480

gtcaaataaa gaaaatagac ctgaagcctc tggcctcccg ctggagtcag agaggacagg      540

cgataacccg actgtgcggg gttccgccgg cgcagatcct gtgggtcagg atgcgcctgg      600

ttggacctgc cagttctgcg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg      660

acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac      720

attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc      780

ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg      840

gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg      900

agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg      960

cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg     1020

ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc     1080

agtcggctcc ctcgttgacc gaatcaccga cctctctccc caggcaagtt tgtacaaaaa     1140

agcaggctgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc     1200

tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg     1260

gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg     1320

tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc     1380

ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg     1440

agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg     1500

agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca     1560

acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg     1620

acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca     1680

gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc     1740

tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc     1800

gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg     1860

agctgtacaa gtaaaaacaa cttgtttatt gcagcttata atggttacaa ataaagcaat     1920

agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc     1980

aaactcatca atgtatctta gccttgcaca gtagtccagc ggtaagggtg tagatcaggc     2040

ccgtctgttt ctcccccgga gctcgctccc ttggcttccc ttatatattt taacatcaga     2100

aacagacatt aaacatctac tgatccaatt tcgccggcgt acggccacga tcgggagggt     2160

gggaatctcg ggggtcttcc gatcctaatc catgatgatt acgacctgag tcactaaaga     2220

cgatggcatg atgatccggc gatgaaaata gccaaatgcc tcgtcatcta attagtga       2278


<210>  50
<211>  2416
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of OMNI50_Scaffold, 106bp homology arm, R2OI_5'RNA, 
       hPGK-GFP-SV40 polyA, R2OI_3'RNA, 30bp - 3' homology arm

<400>  50
gtttgagagt tatgtaagaa attacatgac gagttcaaat aaaaatttat tcaaaccgcc       60

tatttatagg ccgcagatgt tctgcattat gcttgctatt gcaagctttt ttgcgggtgt      120

tgacgcgatg tgatttctgc ccagtgctct gaatgtcaaa gtgaagaaat tcaatgaagc      180

gcgggtaaac ggcgggagta actatgactc tcttaaggcg cacaggggac acagagcctg      240

cccaagtacc gctcccgagg gagcgggaaa cgggggggtg actatcccct ggggtccggc      300

gagagcgctg gtctacggac caggggtggc tgtgggcagg ctgctcctca ggccagttga      360

ttagttacgc atgggctgta cctccacgtg gtcccgctgg taacgacttg tcggctaaat      420

cagcccgccc accatctggg atatggttga ccgtctaacc ccagtactca ggtcacaaac      480

aaaatgggaa cagatacagt gtatgtcggc caggactacc cttctggctt atcaaaacgg      540

gtaccagcac ggttagtggc gggaccgatg ctgcgagagc gaagctgtca cgcccatgtg      600

tttagggctg gacacatgtg gaactggcga accagccttc cgagcgggcg ctgggaccag      660

cccgctttgg agaagtctcg ggtcctaacc cggtcggtgg cgacggccac cgaccccgaa      720

attacctctt acccaggaaa gtccgtatcg acaagtacgc aggttcagga ggaggactgg      780

tgtagccggg agagcgggtg gatctcgcca ggacttgctc ctgaagaacc ctcggtggtg      840

tccgaaatta cagcctccat ggtagcgaca atgagggtag caaccgagga ggtcgtgggg      900

ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt      960

tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc     1020

gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc     1080

ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca     1140

cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg     1200

cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa     1260

ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg     1320

tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga     1380

atcaccgacc tctctcccca ggcaagtttg tacaaaaaag caggctgcca ccatggtgag     1440

caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt     1500

aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct     1560

gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac     1620

caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga     1680

cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga     1740

cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg     1800

catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga     1860

gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa     1920

ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta     1980

ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag     2040

cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga     2100

gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaaaacaact     2160

tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata     2220

aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttagg     2280

gggacagctg ggagtctcgg catgattaca aatcttgcgc tgcactcgga tgtcgtcccc     2340

gtgacggaca cattaatccg gaaagcgagt ggtgactcgc ctcaagtagc caaatgcctc     2400

gtcatctaat tagtga                                                     2416


<210>  51
<211>  2300
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of SPACER, OMNI50_Scaffold, 106bp homology arm, R2_5'RNA, 
       hPGK-GFP-SV40 polyA, R2_3'RNA, 30bp - 3' homology arm

<400>  51
agaagcctga tgttagaatc aagtttgaga gttatgtaag aaattacatg acgagttcaa       60

ataaaaattt attcaaaccg cctatttata ggccgcagat gttctgcatt atgcttgcta      120

ttgcaagctt ttttgcgggt gttgacgcga tgtgatttct gcccagtgct ctgaatgtca      180

aagtgaagaa attcaatgaa gcgcgggtaa acggcgggag taactatgac tctcttaagg      240

tccggaaaca ggtgggccgg ggcgccacca ggggggagca atccctcctg atgatggcga      300

gcaccgcact gtcccttatg ggacggtgta acccggatgg ctgtacacgt ggtaaacacg      360

tgacagcagc cccgatggac ggaccgcgag gaccgtcaag cctagcaggt accttcgggt      420

ggggccttgc gatacctgcg ggcgaaccct gtggtcgggt ttgcagcccg gccacagtgg      480

gtttttttcc tgttgcaaaa aagtcaaata aagaaaatag acctgaagcc tctggcctcc      540

cgctggagtc agagaggaca ggcgataacc cgactgtgcg gggttccgcc ggcgcagatc      600

ctgtgggtca ggatgcgcct ggttggacct gccagttctg cgggttgcgc cttttccaag      660

gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg      720

gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg      780

ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt      840

ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc      900

tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca      960

atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg     1020

cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca     1080

agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc     1140

cccaggcaag tttgtacaaa aaagcaggct gccaccatgg tgagcaaggg cgaggagctg     1200

ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc     1260

agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc     1320

tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc     1380

gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc     1440

atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag     1500

acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc     1560

atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc     1620

cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc     1680

cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc     1740

atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg     1800

agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc     1860

gggatcactc tcggcatgga cgagctgtac aagtaaaaac aacttgttta ttgcagctta     1920

taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact     1980

gcattctagt tgtggtttgt ccaaactcat caatgtatct tagccttgca cagtagtcca     2040

gcggtaaggg tgtagatcag gcccgtctgt ttctcccccg gagctcgctc ccttggcttc     2100

ccttatatat tttaacatca gaaacagaca ttaaacatct actgatccaa tttcgccggc     2160

gtacggccac gatcgggagg gtgggaatct cgggggtctt ccgatcctaa tccatgatga     2220

ttacgacctg agtcactaaa gacgatggca tgatgatccg gcgatgaaaa tagccaaatg     2280

cctcgtcatc taattagtga                                                 2300


<210>  52
<211>  2438
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Fusion of SPACER OMNI50_Scaffold, 106bp homology arm, R2OI_5'RNA,
       hPGK-GFP-SV40 polyA, R2OI_3'RNA, 30bp - 3' homology arm

<400>  52
agaagcctga tgttagaatc aagtttgaga gttatgtaag aaattacatg acgagttcaa       60

ataaaaattt attcaaaccg cctatttata ggccgcagat gttctgcatt atgcttgcta      120

ttgcaagctt ttttgcgggt gttgacgcga tgtgatttct gcccagtgct ctgaatgtca      180

aagtgaagaa attcaatgaa gcgcgggtaa acggcgggag taactatgac tctcttaagg      240

cgcacagggg acacagagcc tgcccaagta ccgctcccga gggagcggga aacggggggg      300

tgactatccc ctggggtccg gcgagagcgc tggtctacgg accaggggtg gctgtgggca      360

ggctgctcct caggccagtt gattagttac gcatgggctg tacctccacg tggtcccgct      420

ggtaacgact tgtcggctaa atcagcccgc ccaccatctg ggatatggtt gaccgtctaa      480

ccccagtact caggtcacaa acaaaatggg aacagataca gtgtatgtcg gccaggacta      540

cccttctggc ttatcaaaac gggtaccagc acggttagtg gcgggaccga tgctgcgaga      600

gcgaagctgt cacgcccatg tgtttagggc tggacacatg tggaactggc gaaccagcct      660

tccgagcggg cgctgggacc agcccgcttt ggagaagtct cgggtcctaa cccggtcggt      720

ggcgacggcc accgaccccg aaattacctc ttacccagga aagtccgtat cgacaagtac      780

gcaggttcag gaggaggact ggtgtagccg ggagagcggg tggatctcgc caggacttgc      840

tcctgaagaa ccctcggtgg tgtccgaaat tacagcctcc atggtagcga caatgagggt      900

agcaaccgag gaggtcgtgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg      960

acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac     1020

attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc     1080

ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg     1140

gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg     1200

agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg     1260

cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg     1320

ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc     1380

agtcggctcc ctcgttgacc gaatcaccga cctctctccc caggcaagtt tgtacaaaaa     1440

agcaggctgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc     1500

tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg     1560

gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg     1620

tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc     1680

ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg     1740

agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg     1800

agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca     1860

acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg     1920

acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca     1980

gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc     2040

tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc     2100

gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg     2160

agctgtacaa gtaaaaacaa cttgtttatt gcagcttata atggttacaa ataaagcaat     2220

agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc     2280

aaactcatca atgtatctta gggggacagc tgggagtctc ggcatgatta caaatcttgc     2340

gctgcactcg gatgtcgtcc ccgtgacgga cacattaatc cggaaagcga gtggtgactc     2400

gcctcaagta gccaaatgcc tcgtcatcta attagtga                             2438


<210>  53
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  nCas9 (D10A) codon optimized for human

<400>  53
atggacaaga agtatagcat cggcctggcc atcggcacaa actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccaagcaag aagtttaagg tgctgggcaa caccgataga      120

cactccatca agaagaatct gatcggcgcc ctgctgttcg actctggcga gacagccgag      180

gccacacggc tgaagagaac cgcccggaga aggtatacac gccggaagaa taggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgattcttt ctttcaccgc      300

ctggaggaga gcttcctggt ggaggaggat aagaagcacg agcggcaccc tatctttggc      360

aacatcgtgg acgaggtggc ctatcacgag aagtacccaa caatctatca cctgaggaag      420

aagctggtgg actccaccga taaggccgac ctgcgcctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca ctttctgatc gagggcgatc tgaacccaga caatagcgat      540

gtggacaagc tgttcatcca gctggtgcag acctacaatc agctgtttga ggagaacccc      600

atcaatgcct ctggagtgga cgcaaaggca atcctgagcg ccagactgtc caagtctaga      660

aggctggaga acctgatcgc ccagctgcca ggcgagaaga agaacggcct gtttggcaat      720

ctgatcgccc tgtccctggg cctgacaccc aacttcaagt ctaattttga tctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tatgacgatg acctggataa cctgctggcc      840

cagatcggcg atcagtacgc cgacctgttc ctggccgcca agaatctgtc tgacgccatc      900

ctgctgagcg atatcctgcg cgtgaacacc gagatcacaa aggcccccct gagcgcctcc      960

atgatcaaga gatatgacga gcaccaccag gatctgaccc tgctgaaggc cctggtgagg     1020

cagcagctgc ctgagaagta caaggagatc ttctttgatc agagcaagaa tggatacgca     1080

ggatatatcg acggaggagc atcccaggag gagttctaca agtttatcaa gcctatcctg     1140

gagaagatgg acggcacaga ggagctgctg gtgaagctga atcgggagga cctgctgagg     1200

aagcagcgca cctttgataa cggcagcatc cctcaccaga tccacctggg agagctgcac     1260

gcaatcctgc gccggcagga ggacttctac ccatttctga aggataaccg ggagaagatc     1320

gagaagatcc tgacattcag aatcccctac tatgtgggac ctctggcccg gggcaatagc     1380

agatttgcct ggatgacccg caagtccgag gagacaatca caccctggaa cttcgaggag     1440

gtggtggata agggcgcctc tgcccagagc ttcatcgagc ggatgaccaa ttttgacaag     1500

aacctgccta atgagaaggt gctgccaaag cactctctgc tgtacgagta tttcaccgtg     1560

tataacgagc tgacaaaggt gaagtacgtg accgagggca tgagaaagcc tgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttta agaccaatag gaaggtgaca     1680

gtgaagcagc tgaaggagga ctatttcaag aagatcgagt gttttgattc tgtggagatc     1740

agcggcgtgg aggacaggtt taacgcctcc ctgggcacct accacgatct gctgaagatc     1800

atcaaggata aggacttcct ggacaacgag gagaatgagg atatcctgga ggacatcgtg     1860

ctgaccctga cactgtttga ggatagggag atgatcgagg agcgcctgaa gacatatgcc     1920

cacctgttcg atgacaaagt gatgaagcag ctgaagagaa ggcgctacac cggatggggc     1980

cggctgagca gaaagctgat caatggcatc cgcgacaagc agtctggcaa gacaatcctg     2040

gactttctga agagcgatgg cttcgccaac cggaacttca tgcagctgat ccacgatgac     2100

tccctgacct tcaaggagga tatccagaag gcacaggtgt ctggacaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggctct cctgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaagtg atgggcaggc acaagccaga gaacatcgtg     2280

atcgagatgg cccgcgagaa tcagaccaca cagaagggcc agaagaactc ccgggagaga     2340

atgaagagaa tcgaggaggg catcaaggag ctgggctctc agatcctgaa ggagcacccc     2400

gtggagaaca cacagctgca gaatgagaag ctgtatctgt actatctgca gaatggccgg     2460

gatatgtacg tggaccagga gctggatatc aacagactgt ctgattatga cgtggatcac     2520

atcgtgccac agtccttcct gaaggatgac tctatcgaca ataaggtgct gaccaggagc     2580

gacaagaacc gcggcaagtc cgataatgtg ccctctgagg aggtggtgaa gaagatgaag     2640

aactactgga ggcagctgct gaatgccaag ctgatcacac agaggaagtt tgataacctg     2700

accaaggcag agaggggagg actgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga caagacagat cacaaagcac gtggcccaga tcctggattc tagaatgaac     2820

acaaagtacg atgagaatga caagctgatc agggaggtga aagtgatcac cctgaagtcc     2880

aagctggtgt ctgactttag gaaggatttc cagttttata aggtgcgcga gatcaacaat     2940

tatcaccacg cccacgacgc ctacctgaac gccgtggtgg gcacagccct gatcaagaag     3000

taccctaagc tggagtccga gttcgtgtac ggcgactata aggtgtacga tgtgcgcaag     3060

atgatcgcca agtctgagca ggagatcggc aaggccaccg ccaagtattt cttttacagc     3120

aacatcatga atttctttaa gaccgagatc acactggcca atggcgagat caggaagcgc     3180

ccactgatcg agacaaacgg cgagacaggc gagatcgtgt gggacaaggg cagggatttt     3240

gccaccgtgc gcaaggtgct gagcatgccc caagtgaata tcgtgaagaa gaccgaggtg     3300

cagacaggcg gcttctccaa ggagtctatc ctgcctaagc ggaactccga taagctgatc     3360

gccagaaaga aggactggga ccccaagaag tatggcggct tcgacagccc tacagtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cgcagctcct tcgagaagaa tcctatcgac     3540

tttctggagg ccaagggcta taaggaggtg aagaaggacc tgatcatcaa gctgccaaag     3600

tactctctgt ttgagctgga gaacggaagg aagagaatgc tggcaagcgc cggagagctg     3660

cagaagggca atgagctggc cctgccctcc aagtacgtga acttcctgta tctggcctcc     3720

cactacgaga agctgaaggg ctctcctgag gataacgagc agaagcagct gtttgtggag     3780

cagcacaagc actatctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgccaatct ggataaggtg ctgtccgcct acaacaagca ccgggataag     3900

ccaatcagag agcaggccga gaatatcatc cacctgttta ccctgacaaa cctgggagca     3960

ccagcagcct tcaagtattt tgacaccaca atcgacagga agcggtacac cagcacaaag     4020

gaggtgctgg acgccacact gatccaccag tccatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgattga                                         4107


<210>  54
<211>  4107
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  nCas9 (N863A) codon optimized for human

<400>  54
atggacaaga agtatagcat cggcctggat atcggcacaa actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccaagcaag aagtttaagg tgctgggcaa caccgataga      120

cactccatca agaagaatct gatcggcgcc ctgctgttcg actctggcga gacagccgag      180

gccacacggc tgaagagaac cgcccggaga aggtatacac gccggaagaa taggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgattcttt ctttcaccgc      300

ctggaggaga gcttcctggt ggaggaggat aagaagcacg agcggcaccc tatctttggc      360

aacatcgtgg acgaggtggc ctatcacgag aagtacccaa caatctatca cctgaggaag      420

aagctggtgg actccaccga taaggccgac ctgcgcctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca ctttctgatc gagggcgatc tgaacccaga caatagcgat      540

gtggacaagc tgttcatcca gctggtgcag acctacaatc agctgtttga ggagaacccc      600

atcaatgcct ctggagtgga cgcaaaggca atcctgagcg ccagactgtc caagtctaga      660

aggctggaga acctgatcgc ccagctgcca ggcgagaaga agaacggcct gtttggcaat      720

ctgatcgccc tgtccctggg cctgacaccc aacttcaagt ctaattttga tctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tatgacgatg acctggataa cctgctggcc      840

cagatcggcg atcagtacgc cgacctgttc ctggccgcca agaatctgtc tgacgccatc      900

ctgctgagcg atatcctgcg cgtgaacacc gagatcacaa aggcccccct gagcgcctcc      960

atgatcaaga gatatgacga gcaccaccag gatctgaccc tgctgaaggc cctggtgagg     1020

cagcagctgc ctgagaagta caaggagatc ttctttgatc agagcaagaa tggatacgca     1080

ggatatatcg acggaggagc atcccaggag gagttctaca agtttatcaa gcctatcctg     1140

gagaagatgg acggcacaga ggagctgctg gtgaagctga atcgggagga cctgctgagg     1200

aagcagcgca cctttgataa cggcagcatc cctcaccaga tccacctggg agagctgcac     1260

gcaatcctgc gccggcagga ggacttctac ccatttctga aggataaccg ggagaagatc     1320

gagaagatcc tgacattcag aatcccctac tatgtgggac ctctggcccg gggcaatagc     1380

agatttgcct ggatgacccg caagtccgag gagacaatca caccctggaa cttcgaggag     1440

gtggtggata agggcgcctc tgcccagagc ttcatcgagc ggatgaccaa ttttgacaag     1500

aacctgccta atgagaaggt gctgccaaag cactctctgc tgtacgagta tttcaccgtg     1560

tataacgagc tgacaaaggt gaagtacgtg accgagggca tgagaaagcc tgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttta agaccaatag gaaggtgaca     1680

gtgaagcagc tgaaggagga ctatttcaag aagatcgagt gttttgattc tgtggagatc     1740

agcggcgtgg aggacaggtt taacgcctcc ctgggcacct accacgatct gctgaagatc     1800

atcaaggata aggacttcct ggacaacgag gagaatgagg atatcctgga ggacatcgtg     1860

ctgaccctga cactgtttga ggatagggag atgatcgagg agcgcctgaa gacatatgcc     1920

cacctgttcg atgacaaagt gatgaagcag ctgaagagaa ggcgctacac cggatggggc     1980

cggctgagca gaaagctgat caatggcatc cgcgacaagc agtctggcaa gacaatcctg     2040

gactttctga agagcgatgg cttcgccaac cggaacttca tgcagctgat ccacgatgac     2100

tccctgacct tcaaggagga tatccagaag gcacaggtgt ctggacaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggctct cctgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaagtg atgggcaggc acaagccaga gaacatcgtg     2280

atcgagatgg cccgcgagaa tcagaccaca cagaagggcc agaagaactc ccgggagaga     2340

atgaagagaa tcgaggaggg catcaaggag ctgggctctc agatcctgaa ggagcacccc     2400

gtggagaaca cacagctgca gaatgagaag ctgtatctgt actatctgca gaatggccgg     2460

gatatgtacg tggaccagga gctggatatc aacagactgt ctgattatga cgtggatcac     2520

atcgtgccac agtccttcct gaaggatgac tctatcgaca ataaggtgct gaccaggagc     2580

gacaaggccc gcggcaagtc cgataatgtg ccctctgagg aggtggtgaa gaagatgaag     2640

aactactgga ggcagctgct gaatgccaag ctgatcacac agaggaagtt tgataacctg     2700

accaaggcag agaggggagg actgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga caagacagat cacaaagcac gtggcccaga tcctggattc tagaatgaac     2820

acaaagtacg atgagaatga caagctgatc agggaggtga aagtgatcac cctgaagtcc     2880

aagctggtgt ctgactttag gaaggatttc cagttttata aggtgcgcga gatcaacaat     2940

tatcaccacg cccacgacgc ctacctgaac gccgtggtgg gcacagccct gatcaagaag     3000

taccctaagc tggagtccga gttcgtgtac ggcgactata aggtgtacga tgtgcgcaag     3060

atgatcgcca agtctgagca ggagatcggc aaggccaccg ccaagtattt cttttacagc     3120

aacatcatga atttctttaa gaccgagatc acactggcca atggcgagat caggaagcgc     3180

ccactgatcg agacaaacgg cgagacaggc gagatcgtgt gggacaaggg cagggatttt     3240

gccaccgtgc gcaaggtgct gagcatgccc caagtgaata tcgtgaagaa gaccgaggtg     3300

cagacaggcg gcttctccaa ggagtctatc ctgcctaagc ggaactccga taagctgatc     3360

gccagaaaga aggactggga ccccaagaag tatggcggct tcgacagccc tacagtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cgcagctcct tcgagaagaa tcctatcgac     3540

tttctggagg ccaagggcta taaggaggtg aagaaggacc tgatcatcaa gctgccaaag     3600

tactctctgt ttgagctgga gaacggaagg aagagaatgc tggcaagcgc cggagagctg     3660

cagaagggca atgagctggc cctgccctcc aagtacgtga acttcctgta tctggcctcc     3720

cactacgaga agctgaaggg ctctcctgag gataacgagc agaagcagct gtttgtggag     3780

cagcacaagc actatctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgccaatct ggataaggtg ctgtccgcct acaacaagca ccgggataag     3900

ccaatcagag agcaggccga gaatatcatc cacctgttta ccctgacaaa cctgggagca     3960

ccagcagcct tcaagtattt tgacaccaca atcgacagga agcggtacac cagcacaaag     4020

gaggtgctgg acgccacact gatccaccag tccatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgattga                                         4107


<210>  55
<211>  4200
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  OMNI50 - nuclease sequence with HA-tag and NLS

<400>  55
atgcctaaga agaagagaaa ggtgggtacc accaaggtga aggactacta cataggcttg       60

gacatcggca cctctagcgt cgggtgggcc gtcaccgatg aagcctataa cgtgcttaag      120

tttaatagca agaaaatgtg gggcgtgcgg ctgttcgacg acgctaagac ggcagaggag      180

cgtaggggcc agcgaggagc aagacgacgt ctggatcgga agaaggagag actcagcctg      240

ctgcaggact tcttcgccga agaggtagca aaggtcgacc ccaacttctt cctcaggctg      300

gacaattccg atctgtacat ggaagataag gaccagaaac tgaaaagcaa atatacactg      360

ttcaacgaca aggacttcaa ggataagaat tttcataaga agtaccccac aatacatcac      420

ctgctgatgg atctgatcga ggacgacagt aagaaggaca tccggctcgt ctacctggcc      480

tgtcactatt tgctcaagaa caggggtcat ttcatcttcg agggccagaa gttcgacact      540

aaatcaagct tcgagaacag tttgaacgag ctcaaagttc atttgaacga cgagtatgga      600

ctggacctcg aatttgacaa cgagaacctg attaacatct tgactgaccc aaaactcaat      660

aaaacggcca agaagaagga gctgaagtcc gtaatcggcg acaccaagtt cctcaaagcc      720

gtttccgcga taatgatcgg ctctagccag aaactcgtcg acttgttcga gaaccccgag      780

gatttcgacg actctgcgat aaagtccgtt gacttctcaa ctacctcttt cgacgacaag      840

tactctgact atgaactcgc tctgggtgac aagatcgctc tggtcaacat ccttaaggaa      900

atttacgata gctccatcct cgagaacctg ctcaaagagg cagacaagtc taaggacggt      960

aacaaatata tcagtaatgc attcgtgaag aagtacaata aacacggaca agatctgaaa     1020

gagttcaaac gtctggtacg acaatatcac aagagtgcgt attttgatat tttcagatcc     1080

gagaaggtga atgacaatta cgtcagctac actaaaagct caattagcaa caataaacgc     1140

gtcaaagcaa acaagttcac tgatcaagag gccttctaca aattcgccaa gaaacatctg     1200

gagacaatca agtataagat caacaaggta aacggctcca aggcagatct ggagctgatt     1260

gacgggatgc tgcgggacat ggagttcaag aactttatgc ccaaaattaa gtccagtgac     1320

aacggggtga ttccatacca gctcaagctg atggaattga acaaaatact cgagaatcag     1380

tcaaagcatc acgagttcct caatgtcagc gacgagtacg gctccgtgtg tgataaaatc     1440

gcatctatca tggagttccg tatcccctac tacgtgggac ccctgaaccc caatagcaag     1500

tacgcctgga tcaagaagca gaaagatagt gagattactc cctggaactt caaggacgtc     1560

gtggacctcg actccagcag agaggagttc attgactcac tgatcggacg ctgtacttac     1620

cttaaggacg agaaggtcct tcccaaagct tctttgctgt ataacgaata catggtgctg     1680

aacgagctga ataacctgaa gttgaacgac cttcccatca ccgaggagat gaagaagaag     1740

atatttgacc agttgttcaa aacaagaaag aaggtcaccc ttaaagcggt ggcaaacctg     1800

ctgaagaagg agttcaacat caacggcgag attctgctct ctgggaccga cggtgacttc     1860

aagcagggct tgaactcata caatgacttc aaagctatcg tgggcgataa agtcgattcc     1920

gatgattacc gggacaagat tgaggagatc attaaactga tagttcttta cggtgacgat     1980

aagagttacc ttcagaagaa gattaaagct gggtatggaa aatacttcac cgacagtgag     2040

attaagaaaa tggcggggct gaactacaag gattggggaa ggctctcaaa gaagctgctg     2100

acgggactcg agggtgcaaa caagatcact ggagagcggg gctccattat tcacttcatg     2160

agggaatata accttaatct gatggagctt atgtcagctt catttacgtt caccgaagag     2220

atacagaaac ttaaccccgt ggatgaccgc aagctgtcat acgaaatggt ggacgaactg     2280

tacctttctc ccagtgtgaa acggatgctc tggcagtccc tgcgcatcgt cgacgagata     2340

aagaacatca tgggaaccga cagtaagaag attttcatcg agatggctcg gggtaaggaa     2400

gaggtgaaag cccgcaagga gtcaaggaag aaccaactgc tgaagttcta taaagacgga     2460

aagaaggcat tcatcagcga gattggcgag gagaggtact cttacttgct ttctgagata     2520

gagggtgagg aagagaataa gtttcgatgg gataacctgt acctttatta tactcaactg     2580

ggtcgctgca tgtactcttt ggaacctatc gacatatctg agctgtcttc aaagaatatt     2640

tacgatcagg atcatatcta ccccaaaagc aagatttacg acgacagtat cgagaatagg     2700

gtgctggtga agaaggacct taactccaag aagggtaaca gctatcctat cccagacgaa     2760

atcctgaaca agaactgtta cgcctactgg aagatcctgt acgataaagg tcttatcggg     2820

cagaagaagt acactcggct gacccggaga actggcttca cggacgacga gctcgttcag     2880

ttcatctcaa gacagatcgt ggaaactaga caagcaacaa aggagactgc taacctgctc     2940

aagacaatat gtaagaactc cgagatcgtg tattccaaag ccgagaacgc aagtcggttt     3000

aggcaagagt tcgacatcgt gaagtgtagg gcggtgaacg atcttcatca tatgcacgat     3060

gcctacatca acatcatagt ggggaacgtg tataacacca agttcacgaa ggaccctatg     3120

aatttcgtaa agaagcagga aaaggcgcgg agctacaatc tcgagaatat gttcaagtac     3180

gatgtgaaac gtggcggata caccgcttgg atcgccgatg acgagaaggg caccgtgaag     3240

aacgcgagta ttaaacgtat ccggaaggag ctggaaggca caaattatag gttcacaaga     3300

atgaactaca ttgagtctgg agcgcttttc aacgccactc tccagcggaa gaataagggc     3360

tccagacccc tgaaggacaa aggcccgaaa tcttccatcg agaagtacgg cggctacaca     3420

aacatcaata aagcctgttt cgctgttctt gacatcaagt ctaagaacaa gattgagagg     3480

aagctgatgc ccgtcgagcg tgagatctat gccaaacaga agaacgacaa gaagctgtcc     3540

gacgagattt tctcaaagta cctcaaggac cgatttggca tcgaggacta cagggttgtc     3600

tacccagtgg tgaaaatgcg cacactgctc aagatcgacg gcagctacta cttcatcaca     3660

ggcggttctg ataagaccct ggagttgcga tctgctctgc agctgattct ccctaagaag     3720

aacgagtggg cgatcaaaca gatcgacaag tcttccgaaa acgactatct gacgatcgag     3780

cgtatccagg acctgaccga ggagctggtg tataacactt tcgacatcat cgtcaacaag     3840

ttcaagacca gtgtcttcaa gaagtctttc cttaacttgt ttcaggacga caagattgag     3900

aacattgact tcaagtttaa gtccatggac ttcaaggaga aatgcaagac acttctcatg     3960

ctggtcaagg cgattcgggc atccggcgtg aggcaggatc tcaagtccat cgacctcaag     4020

tctgattacg gacggctcag ttcaaagacc aacaacatcg gcaattacca ggagttcaag     4080

attattaatc agtccatcac tggactgttc gagaatgagg tcgatctcct gaagctggga     4140

tcctacccat acgatgttcc agattacgcg gccgctccaa aaaagaaaag aaaagtttaa     4200


<210>  56
<211>  3906
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  R2OI (C248S, C251S, W294A) with HA-tag and NLS - R2OI mutant, 
       point mutations in catalytic residues of DNA-binding domain

<400>  56
atgggcaccg acacagtgta cgtcggccag gattatccta gcggcctgag caaaagagtg       60

cccgctagac tggttgctgg ccccatgctg agagagagat cttgtcacgc ccacgtgttc      120

agagccggac acatgtggaa ttggagaacc agcctgccta gcggcagatg ggatcagcct      180

gctctggaaa agtcccgggt gctgaccaga tctgtggcca ccgctacaga ccccgagatc      240

acatcttacc ctggcaagag cgtgtccacc agcacacagg tgcaagaaga ggactggtgt      300

agcagagaga gcggctggat ttctcctgga ctggcccctg aggaacctag cgtggtgtct      360

gagatcacag cctccatggt ggccactatg agagtggcta cagaggaagt ggtgctggaa      420

cctcagcctg agcaggtcgt gacaattctg cccgagcacg gcagaaatgt gccaccagga      480

ctggccgagc aggataccgc ctctcctatt gaagtgtccg tgctgctgcc cgacctggcc      540

gaaaattgtc ctctgtgtgg tgttcccagc ggcggactga gactgctggg aaagcacttt      600

gccgttagac atgccggcgt gcccgtgacc tacgagtgta gaaagtgtgc ctggcggagc      660

cccaatagcc acagcatctc ttgccacgtg ccaaagtgca gaggcagagc cagaatgcca      720

agcggagatc ctggaatcgc cagcgatctg agcgaggcca gatttgccac agaagtggga      780

gtcgcccagc acaagagaca cgtgcacccc gtggaatgga acaaagtgcg gctggaaaga      840

agaggcgcca gaggcggagg aatcaaggcc acaaaacttg ccagcgtggc cgaggtggaa      900

accctgatca gactgattag agagcacggc gatagcggcg ccacatacca gctgattgcc      960

gatgaactcg gcagaggcaa gacagccgag caagtgcgga gcaagaagcg gctgctgaga     1020

atcgataccg ccagcaactc tcccgacgac gccgaagtgg aagaggaaag actggaatct     1080

ctggccgtgc ggtccagcag cagatctcct cctagtctgg tggctaccag agtgcgggaa     1140

gctgtggcaa ggggagaatc tgaaggcggc gaggaaatca gagccattgc cgcactgatc     1200

agagatgtgg atcagaaccc ctgcctgatc gagacaagcg ccagcgacat catcagcaag     1260

ctgggcagaa gagtggacgg ccctaaaaga cccagacctg tcgtgcggga acagacccaa     1320

gaaaaaggct gggtccgacg gctggccaga cggaagagag agtatagaga ggcccagtac     1380

ctgtacagca gagatcaggc aagactggcc gctcagattc tggatggcgc tgcctctcaa     1440

gaatgcgccc tgcctgtgga tcaagtgtac ggcgccttcc gggaaaagtg ggagacagtg     1500

ggacagtttc acggcctggg cgagtttaga acaggcgcta gagccgacaa ctgggagttc     1560

tactctccca tcctggctgc cgaagtcaaa gaaaacctga tgcggatggc caacggcaca     1620

gcccctggac ctgatagaat cagcaagaag gccctgctgg actgggaccc tagaggcgaa     1680

cagctggcta gactgtacac cacatggctg atcggcggcg tgatccccag agtgttcaaa     1740

gagtgtcgga ccaagctgct gcctaagagc agcgatcctg tggaactgca ggatatcgga     1800

ggatggcggc ctgtgacaat cggcagcatg gtcaccagac tgttcagcag aatcctgacc     1860

atgcggctga cccgggcctg tcctatcaat cctagacaga gaggcttcct ggccagcagc     1920

tctggatgtg ccgagaacct gctgatcttc gacgagatcg tgcggcggtc tagaagagat     1980

ggtggaccac tggccgtggt gttcgtggat ttcgccagag ccttcgacag catcagccac     2040

gagcacatcc tgtgtgttct ggaagaaggc ggcctggata gacacgtgat cggcctgatt     2100

cggaacagct acgtggactg tgtgaccaga gtgggctgcg tggaaggcat gacacctcca     2160

atccagatga aggtcggagt gaagcagggc gaccctatga gccctctgct gttcaatctg     2220

gctatggacc ctctgattca caagctggaa acagccggca caggcctgaa gtggggagat     2280

ctgtctatcg ccacactggc cttcgccgat gatctggtgc tggtgtcaga cagcgaagaa     2340

ggcatgggca gatccctggg catcctggaa aaattctgcc agctgaccgg cctgagagtg     2400

cagcctagaa agtgccacgg cttcttcatg gacaagggcg tcgtgaatgg ctgcggcaca     2460

tgggagattt gtggcagccc tatccacatg atcccaccag gcgaatctgt gcgctatctg     2520

ggcgttcaag ttggccctgg aagaggcgtg atggaacccg atctgatccc taccgtgcac     2580

acctggatcg agagaatctc tgaggcccct ctgaagccca gccagagaat gagagtgctg     2640

aatagcttcg ccctgccacg gatcatctat caggctgacc tgggcaaagt gaccgtgaca     2700

aagctggccc agatcgatgg aattgtgcgg aaagccgtga agaagtggct gcatctgagc     2760

cccagcacct gtaatggcct gctgtactcc agaaacagag atggcggact ggggctcctg     2820

aagctggaac gactgattcc tagcgtgcgg accaagagaa tctaccggat gagcagaagc     2880

cccgacatct ggaccagaag aatgaccagc cactccgtgt ccaagagcga ctgggaaatg     2940

ctgtgggtgc aagctggcgg agaaagaggc tctgctcctg ttatgggagc cgtggaagcc     3000

gctcctaccg atgtggaaag atcccctgac taccccgatt ggcggagaga ggaaaatctt     3060

gcttggagcg ccctgagagt tcaaggcgtg ggagctgatc agttcagagg cgatagaacc     3120

tccagcagct ggatcgccga acctgcctct gtgggatttg cccagagaca ttggctggct     3180

gctctggcac ttagagccgg cgtgtaccct accagagagt ttctggccag gggcaaagaa     3240

aagagcggag ccgcctgtag aagatgccct gccagactgg aaagctgcag ccacatcctg     3300

ggccagtgtc ctttcgtgca ggccaacaga atcgcccggc acaacaaagt gtgcgtgctc     3360

ctggcaaccg aggccgagag atttggctgg accgtgatcc gggaattccg gcttgaagat     3420

gctgctggcg ggctgaagat tcccgacctc gtgtgtaaaa aggccgacac cgtgctgatc     3480

gtggacgtga ccgtcagata cgagatggac ggcgagacac tgaagagagc cgccagcgag     3540

aaagtgaagc actatctgcc agtgggccag cagatcaccg acaaagtcgg cggacggtgc     3600

ttcaaagtga tgggctttcc tgtgggcgca agaggcaaat ggccagcctc taacaatacc     3660

gtgctggccg aacttggagt gccagccggc agaatgagga cctttgctag gctggtgtcc     3720

cggcggacac tgctgtatag cctggacatc ctgcgggact tcatgagaga gcctgccgga     3780

agaggtacaa gagtggcact gattccagct gccacaggcg ctgctaacgg atcctaccca     3840

tacgatgttc cagattacgc ggccgctcca aaaaagaaaa gaaaagttga attcggcggc     3900

agctag                                                                3906


<210>  57
<211>  3624
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  R2OI(ZF-Myb) with HA-tag and NLS- R2OI mutant, ZF1 and Myb 
       N-terminal domains deleted

<400>  57
atgggcaccg acacagtgta cgtcggccag gattatccta gcggcctgag caaaagagtg       60

cccgctagac tggttgctgg ccccatgctg agagagagat cttgtcacgc ccacgtgttc      120

agagccggac acatgtggaa ttggagaacc agcctgccta gcggcagatg ggatcagcct      180

gctctggaaa agtcccgggt gctgaccaga tctgtggcca ccgctacaga ccccgagatc      240

acatcttacc ctggcaagag cgtgtccacc agcacacagg tgcaagaaga ggactggtgt      300

agcagagaga gcggctggat ttctcctgga ctggcccctg aggaacctag cgtggtgtct      360

gagatcacag cctccatggt ggccactatg agagtggcta cagaggaagt ggtgctggaa      420

cctcagcctg agcaggtcgt gacaattctg cccgagcacg gcagaaatgt gccaccagga      480

ctggccgagc aggataccgc ctctcctatt gaagtgtccg tgctgctgcc cgacctggcc      540

gaaaattgtc ctctgtgtgg tgttcccagc ggcggactga gactgctggg aaagcacttt      600

gccgttagac atgccggcgt gcccgtgacc tacgagtgta gaaagtgtgc ctggcggagc      660

cccaatagcc acagcatctc ttgccacgtg ccaaagtgca gaggcagagc cagaatgcca      720

agcggagatc tgctgagaat cgataccgcc agcaactctc ccgacgacgc cgaagtggaa      780

gaggaaagac tggaatctct ggccgtgcgg tccagcagca gatctcctcc tagtctggtg      840

gctaccagag tgcgggaagc tgtggcaagg ggagaatctg aaggcggcga ggaaatcaga      900

gccattgccg cactgatcag agatgtggat cagaacccct gcctgatcga gacaagcgcc      960

agcgacatca tcagcaagct gggcagaaga gtggacggcc ctaaaagacc cagacctgtc     1020

gtgcgggaac agacccaaga aaaaggctgg gtccgacggc tggccagacg gaagagagag     1080

tatagagagg cccagtacct gtacagcaga gatcaggcaa gactggccgc tcagattctg     1140

gatggcgctg cctctcaaga atgcgccctg cctgtggatc aagtgtacgg cgccttccgg     1200

gaaaagtggg agacagtggg acagtttcac ggcctgggcg agtttagaac aggcgctaga     1260

gccgacaact gggagttcta ctctcccatc ctggctgccg aagtcaaaga aaacctgatg     1320

cggatggcca acggcacagc ccctggacct gatagaatca gcaagaaggc cctgctggac     1380

tgggacccta gaggcgaaca gctggctaga ctgtacacca catggctgat cggcggcgtg     1440

atccccagag tgttcaaaga gtgtcggacc aagctgctgc ctaagagcag cgatcctgtg     1500

gaactgcagg atatcggagg atggcggcct gtgacaatcg gcagcatggt caccagactg     1560

ttcagcagaa tcctgaccat gcggctgacc cgggcctgtc ctatcaatcc tagacagaga     1620

ggcttcctgg ccagcagctc tggatgtgcc gagaacctgc tgatcttcga cgagatcgtg     1680

cggcggtcta gaagagatgg tggaccactg gccgtggtgt tcgtggattt cgccagagcc     1740

ttcgacagca tcagccacga gcacatcctg tgtgttctgg aagaaggcgg cctggataga     1800

cacgtgatcg gcctgattcg gaacagctac gtggactgtg tgaccagagt gggctgcgtg     1860

gaaggcatga cacctccaat ccagatgaag gtcggagtga agcagggcga ccctatgagc     1920

cctctgctgt tcaatctggc tatggaccct ctgattcaca agctggaaac agccggcaca     1980

ggcctgaagt ggggagatct gtctatcgcc acactggcct tcgccgatga tctggtgctg     2040

gtgtcagaca gcgaagaagg catgggcaga tccctgggca tcctggaaaa attctgccag     2100

ctgaccggcc tgagagtgca gcctagaaag tgccacggct tcttcatgga caagggcgtc     2160

gtgaatggct gcggcacatg ggagatttgt ggcagcccta tccacatgat cccaccaggc     2220

gaatctgtgc gctatctggg cgttcaagtt ggccctggaa gaggcgtgat ggaacccgat     2280

ctgatcccta ccgtgcacac ctggatcgag agaatctctg aggcccctct gaagcccagc     2340

cagagaatga gagtgctgaa tagcttcgcc ctgccacgga tcatctatca ggctgacctg     2400

ggcaaagtga ccgtgacaaa gctggcccag atcgatggaa ttgtgcggaa agccgtgaag     2460

aagtggctgc atctgagccc cagcacctgt aatggcctgc tgtactccag aaacagagat     2520

ggcggactgg ggctcctgaa gctggaacga ctgattccta gcgtgcggac caagagaatc     2580

taccggatga gcagaagccc cgacatctgg accagaagaa tgaccagcca ctccgtgtcc     2640

aagagcgact gggaaatgct gtgggtgcaa gctggcggag aaagaggctc tgctcctgtt     2700

atgggagccg tggaagccgc tcctaccgat gtggaaagat cccctgacta ccccgattgg     2760

cggagagagg aaaatcttgc ttggagcgcc ctgagagttc aaggcgtggg agctgatcag     2820

ttcagaggcg atagaacctc cagcagctgg atcgccgaac ctgcctctgt gggatttgcc     2880

cagagacatt ggctggctgc tctggcactt agagccggcg tgtaccctac cagagagttt     2940

ctggccaggg gcaaagaaaa gagcggagcc gcctgtagaa gatgccctgc cagactggaa     3000

agctgcagcc acatcctggg ccagtgtcct ttcgtgcagg ccaacagaat cgcccggcac     3060

aacaaagtgt gcgtgctcct ggcaaccgag gccgagagat ttggctggac cgtgatccgg     3120

gaattccggc ttgaagatgc tgctggcggg ctgaagattc ccgacctcgt gtgtaaaaag     3180

gccgacaccg tgctgatcgt ggacgtgacc gtcagatacg agatggacgg cgagacactg     3240

aagagagccg ccagcgagaa agtgaagcac tatctgccag tgggccagca gatcaccgac     3300

aaagtcggcg gacggtgctt caaagtgatg ggctttcctg tgggcgcaag aggcaaatgg     3360

ccagcctcta acaataccgt gctggccgaa cttggagtgc cagccggcag aatgaggacc     3420

tttgctaggc tggtgtcccg gcggacactg ctgtatagcc tggacatcct gcgggacttc     3480

atgagagagc ctgccggaag aggtacaaga gtggcactga ttccagctgc cacaggcgct     3540

gctaacggat cctacccata cgatgttcca gattacgcgg ccgctccaaa aaagaaaaga     3600

aaagttgaat tcggcggcag ctag                                            3624


<210>  58
<211>  3405
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  R2(C114S, C117S, R151A, W152A) with HA-tag and NLS - R2 mutant, 
       point mutations in catalytic residues of DNA-binding domain

<400>  58
atgatggcca gcacagccct gtctctgatg ggcagatgca atcccgatgg ctgcacaaga       60

ggcaagcacg tgacagccgc tcctatggat ggacctagag gaccttctag cctggccggc      120

acatttggat ggggacttgc tattcctgcc ggcgagcctt gtggcagagt gtgttctcct      180

gccaccgtgg gattcttccc agtggccaag aagtccaaca aagagaacag acccgaggcc      240

agcggcctgc ctctggaatc tgaaagaacc ggcgataatc ctaccgtgcg gggatctgct      300

ggtgccgatc ctgttggaca agatgcccct ggctggacca gccagttcag cgagagaacc      360

ttcagcacca atagaggcct gggcgtgcac aaaagacggg ctcaccctgt ggaaacaaac      420

accgacgctg cccctatgat ggtcaagaga gccgcccacg gcgaggaaat cgacctgctg      480

gccagaacag aagccagact gctggctgag aggggacagt gttctggcgg agatctgttt      540

ggcgccctgc ctggctttgg aagaaccctg gaagccatca agggccagcg cagaagagag      600

ccttatagag ccctggtgca ggcccacctg gccagatttg gatctcagcc tggacctagc      660

tctggcggat gtagcgccga acctgatttt cggagagcct ctggcgctga agaggccggc      720

gaagaaagat gtgctgagga tgccgccgct tacgatcctt ctgctgtggg ccaaatgagc      780

cctgatgccg ccagagtgct gtctgaactt cttgaaggcg ctggcagacg cagagcctgt      840

agagccatga ggcctaagac cgccggaaga agaaacgacc tgcacgacga tagaaccgcc      900

agcgctcaca agaccagcag acagaagaga agggccgagt acgccagggt gcaagagctg      960

tacaagaagt gcagatccag agccgccgct gaagtgattg atggtgcttg tggtggcgtg     1020

ggccacagcc tggaagagat ggaaacctat tggcggccca tcctggaaag agtgtctgac     1080

gctcctggac caacacctga agctctgcat gctctgggca gagctgagtg gcatggcggc     1140

aatagagatt acacccagct gtggaagccc atcagcgtgg aagaaatcaa ggccagcaga     1200

ttcgactggc ggacaagccc tggacctgat ggcattagat ctggacagtg gcgggctgtg     1260

cctgtgcacc tgaaggccga aatgttcaac gcctggatgg ccagaggcga gatccctgag     1320

atcctgagac agtgcagaac cgtgttcgtg cccaaggtgg aaagacctgg cggaccaggc     1380

gagtacagac ccatctctat cgccagcatt cctctgcggc acttccactc tatcctggct     1440

cggagacttc tggcctgctg tcctcctgat gccagacaga gaggctttat ctgcgccgac     1500

ggcaccctgg aaaattctgc agtgctggat gccgtgctgg gcgactctcg gaagaaactg     1560

agagaatgtc acgtggccgt cctggacttc gccaaggcct ttgatacagt gtctcacgag     1620

gccctggtgg aactgctgag actgagggga atgcctgagc agttctgtgg ctatatcgcc     1680

cacctgtacg acaccgcctc taccacactg gccgtgaaca atgagatgag cagccccgtg     1740

aaagttggca gaggcgttag acagggcgac cctctgagcc ccatcctgtt caatgtggtc     1800

atggatctga tcctggccag cctgcctgag agagtgggct atagactgga aatggaactg     1860

gtgtctgccc tggcctacgc cgatgatctg gttctgcttg ccggcagcaa agtgggcatg     1920

caagagtcta tcagcgccgt ggattgcgtg ggcagacaga tgggcctgcg cctgaattgc     1980

agaaaaagcg ccgtgctgag catgatcccc gatggccaca gaaagaagca ccactacctg     2040

accgagcgga ccttcaatat cggcggcaag cctctgagac aggtgtcctg tgttgagaga     2100

tggcggtatc tgggcgtcga ctttgaggcc tctggctgtg tgacactgga acactctatc     2160

agcagcgccc tgaacaacat cagcagagcc cctctgaagc ctcagcagcg gctggaaatt     2220

ctgagagccc atctgatccc tcggttccag cacggatttg tgctgggcaa catctccgac     2280

gaccggctga gaatgctgga cgtgcagatc agaaaagccg tcggccagtg gctgagactt     2340

cctgcagatg tgcctaaggc ctactatcac gctgctgtgc aagatggcgg cctggctatt     2400

ccttctgtgc gcgccacaat tcccgacctg atcgtgcgaa gattcggcgg acttgatagc     2460

tctccttgga gcgtggccag agctgccgcc aagagcgata agatccggaa aaagctgcgc     2520

tgggcctgga agcagctgcg gagattttct agagtggaca gcaccacaca gcggcctagt     2580

gtgcggctgt tttggagaga acatctgcac gcctccgtgg acggcagaga gctgagagaa     2640

agcaccagaa cacccaccag caccaagtgg atcagagaga gatgcgccca gatcacaggc     2700

cgggatttcg tgcagttcgt gcacacccat atcaacgccc tgccatccag aatcaggggc     2760

agcagaggta gaagaggcgg aggcgaaagc agcctgacat gtagagccgg ctgtaaagtg     2820

cgcgagacaa cagcccacat cctgcagcag tgtcatagaa cacacggcgg cagaatcctg     2880

cggcacaaca agattgtgtc cttcgtggcc aaggccatgg aagagaacaa gtggaccgtg     2940

gaactggaac ccagactgag aacaagcgtg ggcctgagaa agcccgacat cattgcctct     3000

cgagatggcg tgggagtgat cgtggatgtg caggttgtgt caggccagag aagcctggac     3060

gagctgcaca gagagaagcg gaacaaatac ggcaaccacg gcgagctggt tgaactggtt     3120

gcaggcagac tgggactgcc aaaagccgag tgtgtgcggg ccacctcttg taccatttct     3180

tggagaggcg tgtggtccct gaccagctac aaagagctgc ggtccatcat cggactgaga     3240

gagcctacac tgcagatcgt ccccattctg gccctgagag gcagccacat gaattggacc     3300

cgcttcaacc agatgaccag cgtgatggga ggcggcgttg gaggatccta cccatacgat     3360

gttccagatt acgcggccgc tccaaaaaag aaaagaaaag tttag                     3405


<210>  59
<211>  3138
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  R2(ZF-Myb) with HA-tag and NLS - R2 mutant, ZF1 and Myb 
       N-terminal domains deleted

<400>  59
atgatggcca gcacagccct gtctctgatg ggcagatgca atcccgatgg ctgcacaaga       60

ggcaagcacg tgacagccgc tcctatggat ggacctagag gaccttctag cctggccggc      120

acatttggat ggggacttgc tattcctgcc ggcgagcctt gtggcagagt gtgttctcct      180

gccaccgtgg gattcttccc agtggccaag aagtccaaca aagagaacag acccgaggcc      240

agcggcctgc ctctggaatc tgaaagaacc ggcgataatc ctaccgtgcg gggatctgct      300

ggtgccgatc ctgttggaca agatgccaga gagccttata gagccctggt gcaggcccac      360

ctggccagat ttggatctca gcctggacct agctctggcg gatgtagcgc cgaacctgat      420

tttcggagag cctctggcgc tgaagaggcc ggcgaagaaa gatgtgctga ggatgccgcc      480

gcttacgatc cttctgctgt gggccaaatg agccctgatg ccgccagagt gctgtctgaa      540

cttcttgaag gcgctggcag acgcagagcc tgtagagcca tgaggcctaa gaccgccgga      600

agaagaaacg acctgcacga cgatagaacc gccagcgctc acaagaccag cagacagaag      660

agaagggccg agtacgccag ggtgcaagag ctgtacaaga agtgcagatc cagagccgcc      720

gctgaagtga ttgatggtgc ttgtggtggc gtgggccaca gcctggaaga gatggaaacc      780

tattggcggc ccatcctgga aagagtgtct gacgctcctg gaccaacacc tgaagctctg      840

catgctctgg gcagagctga gtggcatggc ggcaatagag attacaccca gctgtggaag      900

cccatcagcg tggaagaaat caaggccagc agattcgact ggcggacaag ccctggacct      960

gatggcatta gatctggaca gtggcgggct gtgcctgtgc acctgaaggc cgaaatgttc     1020

aacgcctgga tggccagagg cgagatccct gagatcctga gacagtgcag aaccgtgttc     1080

gtgcccaagg tggaaagacc tggcggacca ggcgagtaca gacccatctc tatcgccagc     1140

attcctctgc ggcacttcca ctctatcctg gctcggagac ttctggcctg ctgtcctcct     1200

gatgccagac agagaggctt tatctgcgcc gacggcaccc tggaaaattc tgcagtgctg     1260

gatgccgtgc tgggcgactc tcggaagaaa ctgagagaat gtcacgtggc cgtcctggac     1320

ttcgccaagg cctttgatac agtgtctcac gaggccctgg tggaactgct gagactgagg     1380

ggaatgcctg agcagttctg tggctatatc gcccacctgt acgacaccgc ctctaccaca     1440

ctggccgtga acaatgagat gagcagcccc gtgaaagttg gcagaggcgt tagacagggc     1500

gaccctctga gccccatcct gttcaatgtg gtcatggatc tgatcctggc cagcctgcct     1560

gagagagtgg gctatagact ggaaatggaa ctggtgtctg ccctggccta cgccgatgat     1620

ctggttctgc ttgccggcag caaagtgggc atgcaagagt ctatcagcgc cgtggattgc     1680

gtgggcagac agatgggcct gcgcctgaat tgcagaaaaa gcgccgtgct gagcatgatc     1740

cccgatggcc acagaaagaa gcaccactac ctgaccgagc ggaccttcaa tatcggcggc     1800

aagcctctga gacaggtgtc ctgtgttgag agatggcggt atctgggcgt cgactttgag     1860

gcctctggct gtgtgacact ggaacactct atcagcagcg ccctgaacaa catcagcaga     1920

gcccctctga agcctcagca gcggctggaa attctgagag cccatctgat ccctcggttc     1980

cagcacggat ttgtgctggg caacatctcc gacgaccggc tgagaatgct ggacgtgcag     2040

atcagaaaag ccgtcggcca gtggctgaga cttcctgcag atgtgcctaa ggcctactat     2100

cacgctgctg tgcaagatgg cggcctggct attccttctg tgcgcgccac aattcccgac     2160

ctgatcgtgc gaagattcgg cggacttgat agctctcctt ggagcgtggc cagagctgcc     2220

gccaagagcg ataagatccg gaaaaagctg cgctgggcct ggaagcagct gcggagattt     2280

tctagagtgg acagcaccac acagcggcct agtgtgcggc tgttttggag agaacatctg     2340

cacgcctccg tggacggcag agagctgaga gaaagcacca gaacacccac cagcaccaag     2400

tggatcagag agagatgcgc ccagatcaca ggccgggatt tcgtgcagtt cgtgcacacc     2460

catatcaacg ccctgccatc cagaatcagg ggcagcagag gtagaagagg cggaggcgaa     2520

agcagcctga catgtagagc cggctgtaaa gtgcgcgaga caacagccca catcctgcag     2580

cagtgtcata gaacacacgg cggcagaatc ctgcggcaca acaagattgt gtccttcgtg     2640

gccaaggcca tggaagagaa caagtggacc gtggaactgg aacccagact gagaacaagc     2700

gtgggcctga gaaagcccga catcattgcc tctcgagatg gcgtgggagt gatcgtggat     2760

gtgcaggttg tgtcaggcca gagaagcctg gacgagctgc acagagagaa gcggaacaaa     2820

tacggcaacc acggcgagct ggttgaactg gttgcaggca gactgggact gccaaaagcc     2880

gagtgtgtgc gggccacctc ttgtaccatt tcttggagag gcgtgtggtc cctgaccagc     2940

tacaaagagc tgcggtccat catcggactg agagagccta cactgcagat cgtccccatt     3000

ctggccctga gaggcagcca catgaattgg acccgcttca accagatgac cagcgtgatg     3060

ggaggcggcg ttggaggatc ctacccatac gatgttccag attacgcggc cgctccaaaa     3120

aagaaaagaa aagtttag                                                   3138


<210>  60
<211>  7857
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  deadCas9-NLS-32aa-R2OI(ZF-Myb)-HA-NLS

<400>  60
atggacaaga agtatagcat cggcctggcc atcggcacaa actccgtggg ctgggccgtg       60

atcaccgacg agtacaaggt gccaagcaag aagtttaagg tgctgggcaa caccgataga      120

cactccatca agaagaatct gatcggcgcc ctgctgttcg actctggcga gacagccgag      180

gccacacggc tgaagagaac cgcccggaga aggtatacac gccggaagaa taggatctgc      240

tacctgcagg agatcttcag caacgagatg gccaaggtgg acgattcttt ctttcaccgc      300

ctggaggaga gcttcctggt ggaggaggat aagaagcacg agcggcaccc tatctttggc      360

aacatcgtgg acgaggtggc ctatcacgag aagtacccaa caatctatca cctgaggaag      420

aagctggtgg actccaccga taaggccgac ctgcgcctga tctatctggc cctggcccac      480

atgatcaagt tccggggcca ctttctgatc gagggcgatc tgaacccaga caatagcgat      540

gtggacaagc tgttcatcca gctggtgcag acctacaatc agctgtttga ggagaacccc      600

atcaatgcct ctggagtgga cgcaaaggca atcctgagcg ccagactgtc caagtctaga      660

aggctggaga acctgatcgc ccagctgcca ggcgagaaga agaacggcct gtttggcaat      720

ctgatcgccc tgtccctggg cctgacaccc aacttcaagt ctaattttga tctggccgag      780

gacgccaagc tgcagctgtc caaggacacc tatgacgatg acctggataa cctgctggcc      840

cagatcggcg atcagtacgc cgacctgttc ctggccgcca agaatctgtc tgacgccatc      900

ctgctgagcg atatcctgcg cgtgaacacc gagatcacaa aggcccccct gagcgcctcc      960

atgatcaaga gatatgacga gcaccaccag gatctgaccc tgctgaaggc cctggtgagg     1020

cagcagctgc ctgagaagta caaggagatc ttctttgatc agagcaagaa tggatacgca     1080

ggatatatcg acggaggagc atcccaggag gagttctaca agtttatcaa gcctatcctg     1140

gagaagatgg acggcacaga ggagctgctg gtgaagctga atcgggagga cctgctgagg     1200

aagcagcgca cctttgataa cggcagcatc cctcaccaga tccacctggg agagctgcac     1260

gcaatcctgc gccggcagga ggacttctac ccatttctga aggataaccg ggagaagatc     1320

gagaagatcc tgacattcag aatcccctac tatgtgggac ctctggcccg gggcaatagc     1380

agatttgcct ggatgacccg caagtccgag gagacaatca caccctggaa cttcgaggag     1440

gtggtggata agggcgcctc tgcccagagc ttcatcgagc ggatgaccaa ttttgacaag     1500

aacctgccta atgagaaggt gctgccaaag cactctctgc tgtacgagta tttcaccgtg     1560

tataacgagc tgacaaaggt gaagtacgtg accgagggca tgagaaagcc tgccttcctg     1620

agcggcgagc agaagaaggc catcgtggac ctgctgttta agaccaatag gaaggtgaca     1680

gtgaagcagc tgaaggagga ctatttcaag aagatcgagt gttttgattc tgtggagatc     1740

agcggcgtgg aggacaggtt taacgcctcc ctgggcacct accacgatct gctgaagatc     1800

atcaaggata aggacttcct ggacaacgag gagaatgagg atatcctgga ggacatcgtg     1860

ctgaccctga cactgtttga ggatagggag atgatcgagg agcgcctgaa gacatatgcc     1920

cacctgttcg atgacaaagt gatgaagcag ctgaagagaa ggcgctacac cggatggggc     1980

cggctgagca gaaagctgat caatggcatc cgcgacaagc agtctggcaa gacaatcctg     2040

gactttctga agagcgatgg cttcgccaac cggaacttca tgcagctgat ccacgatgac     2100

tccctgacct tcaaggagga tatccagaag gcacaggtgt ctggacaggg cgacagcctg     2160

cacgagcaca tcgccaacct ggccggctct cctgccatca agaagggcat cctgcagacc     2220

gtgaaggtgg tggacgagct ggtgaaagtg atgggcaggc acaagccaga gaacatcgtg     2280

atcgagatgg cccgcgagaa tcagaccaca cagaagggcc agaagaactc ccgggagaga     2340

atgaagagaa tcgaggaggg catcaaggag ctgggctctc agatcctgaa ggagcacccc     2400

gtggagaaca cacagctgca gaatgagaag ctgtatctgt actatctgca gaatggccgg     2460

gatatgtacg tggaccagga gctggatatc aacagactgt ctgattatga cgtggatcac     2520

atcgtgccac agtccttcct gaaggatgac tctatcgaca ataaggtgct gaccaggagc     2580

gacaaggccc gcggcaagtc cgataatgtg ccctctgagg aggtggtgaa gaagatgaag     2640

aactactgga ggcagctgct gaatgccaag ctgatcacac agaggaagtt tgataacctg     2700

accaaggcag agaggggagg actgtccgag ctggacaagg ccggcttcat caagcggcag     2760

ctggtggaga caagacagat cacaaagcac gtggcccaga tcctggattc tagaatgaac     2820

acaaagtacg atgagaatga caagctgatc agggaggtga aagtgatcac cctgaagtcc     2880

aagctggtgt ctgactttag gaaggatttc cagttttata aggtgcgcga gatcaacaat     2940

tatcaccacg cccacgacgc ctacctgaac gccgtggtgg gcacagccct gatcaagaag     3000

taccctaagc tggagtccga gttcgtgtac ggcgactata aggtgtacga tgtgcgcaag     3060

atgatcgcca agtctgagca ggagatcggc aaggccaccg ccaagtattt cttttacagc     3120

aacatcatga atttctttaa gaccgagatc acactggcca atggcgagat caggaagcgc     3180

ccactgatcg agacaaacgg cgagacaggc gagatcgtgt gggacaaggg cagggatttt     3240

gccaccgtgc gcaaggtgct gagcatgccc caagtgaata tcgtgaagaa gaccgaggtg     3300

cagacaggcg gcttctccaa ggagtctatc ctgcctaagc ggaactccga taagctgatc     3360

gccagaaaga aggactggga ccccaagaag tatggcggct tcgacagccc tacagtggcc     3420

tactccgtgc tggtggtggc caaggtggag aagggcaaga gcaagaagct gaagtccgtg     3480

aaggagctgc tgggcatcac catcatggag cgcagctcct tcgagaagaa tcctatcgac     3540

tttctggagg ccaagggcta taaggaggtg aagaaggacc tgatcatcaa gctgccaaag     3600

tactctctgt ttgagctgga gaacggaagg aagagaatgc tggcaagcgc cggagagctg     3660

cagaagggca atgagctggc cctgccctcc aagtacgtga acttcctgta tctggcctcc     3720

cactacgaga agctgaaggg ctctcctgag gataacgagc agaagcagct gtttgtggag     3780

cagcacaagc actatctgga cgagatcatc gagcagatca gcgagttctc caagagagtg     3840

atcctggccg acgccaatct ggataaggtg ctgtccgcct acaacaagca ccgggataag     3900

ccaatcagag agcaggccga gaatatcatc cacctgttta ccctgacaaa cctgggagca     3960

ccagcagcct tcaagtattt tgacaccaca atcgacagga agcggtacac cagcacaaag     4020

gaggtgctgg acgccacact gatccaccag tccatcaccg gcctgtacga gacacggatc     4080

gacctgtctc agctgggagg cgatggctcc ccaaaaaaga aaagaaaagt tgctagctct     4140

ggtggttctt ctggtggttc tagcggcagc gagactcccg ggacctcaga gtccgccaca     4200

cccgaaagtt ctggtggttc ttctggtggt tctatgggca ccgacacagt gtacgtcggc     4260

caggattatc ctagcggcct gagcaaaaga gtgcccgcta gactggttgc tggccccatg     4320

ctgagagaga gatcttgtca cgcccacgtg ttcagagccg gacacatgtg gaattggaga     4380

accagcctgc ctagcggcag atgggatcag cctgctctgg aaaagtcccg ggtgctgacc     4440

agatctgtgg ccaccgctac agaccccgag atcacatctt accctggcaa gagcgtgtcc     4500

accagcacac aggtgcaaga agaggactgg tgtagcagag agagcggctg gatttctcct     4560

ggactggccc ctgaggaacc tagcgtggtg tctgagatca cagcctccat ggtggccact     4620

atgagagtgg ctacagagga agtggtgctg gaacctcagc ctgagcaggt cgtgacaatt     4680

ctgcccgagc acggcagaaa tgtgccacca ggactggccg agcaggatac cgcctctcct     4740

attgaagtgt ccgtgctgct gcccgacctg gccgaaaatt gtcctctgtg tggtgttccc     4800

agcggcggac tgagactgct gggaaagcac tttgccgtta gacatgccgg cgtgcccgtg     4860

acctacgagt gtagaaagtg tgcctggcgg agccccaata gccacagcat ctcttgccac     4920

gtgccaaagt gcagaggcag agccagaatg ccaagcggag atctgctgag aatcgatacc     4980

gccagcaact ctcccgacga cgccgaagtg gaagaggaaa gactggaatc tctggccgtg     5040

cggtccagca gcagatctcc tcctagtctg gtggctacca gagtgcggga agctgtggca     5100

aggggagaat ctgaaggcgg cgaggaaatc agagccattg ccgcactgat cagagatgtg     5160

gatcagaacc cctgcctgat cgagacaagc gccagcgaca tcatcagcaa gctgggcaga     5220

agagtggacg gccctaaaag acccagacct gtcgtgcggg aacagaccca agaaaaaggc     5280

tgggtccgac ggctggccag acggaagaga gagtatagag aggcccagta cctgtacagc     5340

agagatcagg caagactggc cgctcagatt ctggatggcg ctgcctctca agaatgcgcc     5400

ctgcctgtgg atcaagtgta cggcgccttc cgggaaaagt gggagacagt gggacagttt     5460

cacggcctgg gcgagtttag aacaggcgct agagccgaca actgggagtt ctactctccc     5520

atcctggctg ccgaagtcaa agaaaacctg atgcggatgg ccaacggcac agcccctgga     5580

cctgatagaa tcagcaagaa ggccctgctg gactgggacc ctagaggcga acagctggct     5640

agactgtaca ccacatggct gatcggcggc gtgatcccca gagtgttcaa agagtgtcgg     5700

accaagctgc tgcctaagag cagcgatcct gtggaactgc aggatatcgg aggatggcgg     5760

cctgtgacaa tcggcagcat ggtcaccaga ctgttcagca gaatcctgac catgcggctg     5820

acccgggcct gtcctatcaa tcctagacag agaggcttcc tggccagcag ctctggatgt     5880

gccgagaacc tgctgatctt cgacgagatc gtgcggcggt ctagaagaga tggtggacca     5940

ctggccgtgg tgttcgtgga tttcgccaga gccttcgaca gcatcagcca cgagcacatc     6000

ctgtgtgttc tggaagaagg cggcctggat agacacgtga tcggcctgat tcggaacagc     6060

tacgtggact gtgtgaccag agtgggctgc gtggaaggca tgacacctcc aatccagatg     6120

aaggtcggag tgaagcaggg cgaccctatg agccctctgc tgttcaatct ggctatggac     6180

cctctgattc acaagctgga aacagccggc acaggcctga agtggggaga tctgtctatc     6240

gccacactgg ccttcgccga tgatctggtg ctggtgtcag acagcgaaga aggcatgggc     6300

agatccctgg gcatcctgga aaaattctgc cagctgaccg gcctgagagt gcagcctaga     6360

aagtgccacg gcttcttcat ggacaagggc gtcgtgaatg gctgcggcac atgggagatt     6420

tgtggcagcc ctatccacat gatcccacca ggcgaatctg tgcgctatct gggcgttcaa     6480

gttggccctg gaagaggcgt gatggaaccc gatctgatcc ctaccgtgca cacctggatc     6540

gagagaatct ctgaggcccc tctgaagccc agccagagaa tgagagtgct gaatagcttc     6600

gccctgccac ggatcatcta tcaggctgac ctgggcaaag tgaccgtgac aaagctggcc     6660

cagatcgatg gaattgtgcg gaaagccgtg aagaagtggc tgcatctgag ccccagcacc     6720

tgtaatggcc tgctgtactc cagaaacaga gatggcggac tggggctcct gaagctggaa     6780

cgactgattc ctagcgtgcg gaccaagaga atctaccgga tgagcagaag ccccgacatc     6840

tggaccagaa gaatgaccag ccactccgtg tccaagagcg actgggaaat gctgtgggtg     6900

caagctggcg gagaaagagg ctctgctcct gttatgggag ccgtggaagc cgctcctacc     6960

gatgtggaaa gatcccctga ctaccccgat tggcggagag aggaaaatct tgcttggagc     7020

gccctgagag ttcaaggcgt gggagctgat cagttcagag gcgatagaac ctccagcagc     7080

tggatcgccg aacctgcctc tgtgggattt gcccagagac attggctggc tgctctggca     7140

cttagagccg gcgtgtaccc taccagagag tttctggcca ggggcaaaga aaagagcgga     7200

gccgcctgta gaagatgccc tgccagactg gaaagctgca gccacatcct gggccagtgt     7260

cctttcgtgc aggccaacag aatcgcccgg cacaacaaag tgtgcgtgct cctggcaacc     7320

gaggccgaga gatttggctg gaccgtgatc cgggaattcc ggcttgaaga tgctgctggc     7380

gggctgaaga ttcccgacct cgtgtgtaaa aaggccgaca ccgtgctgat cgtggacgtg     7440

accgtcagat acgagatgga cggcgagaca ctgaagagag ccgccagcga gaaagtgaag     7500

cactatctgc cagtgggcca gcagatcacc gacaaagtcg gcggacggtg cttcaaagtg     7560

atgggctttc ctgtgggcgc aagaggcaaa tggccagcct ctaacaatac cgtgctggcc     7620

gaacttggag tgccagccgg cagaatgagg acctttgcta ggctggtgtc ccggcggaca     7680

ctgctgtata gcctggacat cctgcgggac ttcatgagag agcctgccgg aagaggtaca     7740

agagtggcac tgattccagc tgccacaggc gctgctaacg gatcctaccc atacgatgtt     7800

ccagattacg cggccgctcc aaaaaagaaa agaaaagttg aattcggcgg cagctag        7857


<210>  61
<211>  8094
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  NLS-R2OI (C248S, C251S, W294A)-HA-NLS-XTEN-deadCas9

<400>  61
gccaccatgc ctaagaagaa gagaaaggtg ggtaccatgg gcaccgacac agtgtacgtc       60

ggccaggatt atcctagcgg cctgagcaaa agagtgcccg ctagactggt tgctggcccc      120

atgctgagag agagatcttg tcacgcccac gtgttcagag ccggacacat gtggaattgg      180

agaaccagcc tgcctagcgg cagatgggat cagcctgctc tggaaaagtc ccgggtgctg      240

accagatctg tggccaccgc tacagacccc gagatcacat cttaccctgg caagagcgtg      300

tccaccagca cacaggtgca agaagaggac tggtgtagca gagagagcgg ctggatttct      360

cctggactgg cccctgagga acctagcgtg gtgtctgaga tcacagcctc catggtggcc      420

actatgagag tggctacaga ggaagtggtg ctggaacctc agcctgagca ggtcgtgaca      480

attctgcccg agcacggcag aaatgtgcca ccaggactgg ccgagcagga taccgcctct      540

cctattgaag tgtccgtgct gctgcccgac ctggccgaaa attgtcctct gtgtggtgtt      600

cccagcggcg gactgagact gctgggaaag cactttgccg ttagacatgc cggcgtgccc      660

gtgacctacg agtgtagaaa gtgtgcctgg cggagcccca atagccacag catctcttgc      720

cacgtgccaa agtgcagagg cagagccaga atgccaagcg gagatcctgg aatcgccagc      780

gatctgagcg aggccagatt tgccacagaa gtgggagtcg cccagcacaa gagacacgtg      840

caccccgtgg aatggaacaa agtgcggctg gaaagaagag gcgccagagg cggaggaatc      900

aaggccacaa aacttgccag cgtggccgag gtggaaaccc tgatcagact gattagagag      960

cacggcgata gcggcgccac ataccagctg attgccgatg aactcggcag aggcaagaca     1020

gccgagcaag tgcggagcaa gaagcggctg ctgagaatcg ataccgccag caactctccc     1080

gacgacgccg aagtggaaga ggaaagactg gaatctctgg ccgtgcggtc cagcagcaga     1140

tctcctccta gtctggtggc taccagagtg cgggaagctg tggcaagggg agaatctgaa     1200

ggcggcgagg aaatcagagc cattgccgca ctgatcagag atgtggatca gaacccctgc     1260

ctgatcgaga caagcgccag cgacatcatc agcaagctgg gcagaagagt ggacggccct     1320

aaaagaccca gacctgtcgt gcgggaacag acccaagaaa aaggctgggt ccgacggctg     1380

gccagacgga agagagagta tagagaggcc cagtacctgt acagcagaga tcaggcaaga     1440

ctggccgctc agattctgga tggcgctgcc tctcaagaat gcgccctgcc tgtggatcaa     1500

gtgtacggcg ccttccggga aaagtgggag acagtgggac agtttcacgg cctgggcgag     1560

tttagaacag gcgctagagc cgacaactgg gagttctact ctcccatcct ggctgccgaa     1620

gtcaaagaaa acctgatgcg gatggccaac ggcacagccc ctggacctga tagaatcagc     1680

aagaaggccc tgctggactg ggaccctaga ggcgaacagc tggctagact gtacaccaca     1740

tggctgatcg gcggcgtgat ccccagagtg ttcaaagagt gtcggaccaa gctgctgcct     1800

aagagcagcg atcctgtgga actgcaggat atcggaggat ggcggcctgt gacaatcggc     1860

agcatggtca ccagactgtt cagcagaatc ctgaccatgc ggctgacccg ggcctgtcct     1920

atcaatccta gacagagagg cttcctggcc agcagctctg gatgtgccga gaacctgctg     1980

atcttcgacg agatcgtgcg gcggtctaga agagatggtg gaccactggc cgtggtgttc     2040

gtggatttcg ccagagcctt cgacagcatc agccacgagc acatcctgtg tgttctggaa     2100

gaaggcggcc tggatagaca cgtgatcggc ctgattcgga acagctacgt ggactgtgtg     2160

accagagtgg gctgcgtgga aggcatgaca cctccaatcc agatgaaggt cggagtgaag     2220

cagggcgacc ctatgagccc tctgctgttc aatctggcta tggaccctct gattcacaag     2280

ctggaaacag ccggcacagg cctgaagtgg ggagatctgt ctatcgccac actggccttc     2340

gccgatgatc tggtgctggt gtcagacagc gaagaaggca tgggcagatc cctgggcatc     2400

ctggaaaaat tctgccagct gaccggcctg agagtgcagc ctagaaagtg ccacggcttc     2460

ttcatggaca agggcgtcgt gaatggctgc ggcacatggg agatttgtgg cagccctatc     2520

cacatgatcc caccaggcga atctgtgcgc tatctgggcg ttcaagttgg ccctggaaga     2580

ggcgtgatgg aacccgatct gatccctacc gtgcacacct ggatcgagag aatctctgag     2640

gcccctctga agcccagcca gagaatgaga gtgctgaata gcttcgccct gccacggatc     2700

atctatcagg ctgacctggg caaagtgacc gtgacaaagc tggcccagat cgatggaatt     2760

gtgcggaaag ccgtgaagaa gtggctgcat ctgagcccca gcacctgtaa tggcctgctg     2820

tactccagaa acagagatgg cggactgggg ctcctgaagc tggaacgact gattcctagc     2880

gtgcggacca agagaatcta ccggatgagc agaagccccg acatctggac cagaagaatg     2940

accagccact ccgtgtccaa gagcgactgg gaaatgctgt gggtgcaagc tggcggagaa     3000

agaggctctg ctcctgttat gggagccgtg gaagccgctc ctaccgatgt ggaaagatcc     3060

cctgactacc ccgattggcg gagagaggaa aatcttgctt ggagcgccct gagagttcaa     3120

ggcgtgggag ctgatcagtt cagaggcgat agaacctcca gcagctggat cgccgaacct     3180

gcctctgtgg gatttgccca gagacattgg ctggctgctc tggcacttag agccggcgtg     3240

taccctacca gagagtttct ggccaggggc aaagaaaaga gcggagccgc ctgtagaaga     3300

tgccctgcca gactggaaag ctgcagccac atcctgggcc agtgtccttt cgtgcaggcc     3360

aacagaatcg cccggcacaa caaagtgtgc gtgctcctgg caaccgaggc cgagagattt     3420

ggctggaccg tgatccggga attccggctt gaagatgctg ctggcgggct gaagattccc     3480

gacctcgtgt gtaaaaaggc cgacaccgtg ctgatcgtgg acgtgaccgt cagatacgag     3540

atggacggcg agacactgaa gagagccgcc agcgagaaag tgaagcacta tctgccagtg     3600

ggccagcaga tcaccgacaa agtcggcgga cggtgcttca aagtgatggg ctttcctgtg     3660

ggcgcaagag gcaaatggcc agcctctaac aataccgtgc tggccgaact tggagtgcca     3720

gccggcagaa tgaggacctt tgctaggctg gtgtcccggc ggacactgct gtatagcctg     3780

gacatcctgc gggacttcat gagagagcct gccggaagag gtacaagagt ggcactgatt     3840

ccagctgcca caggcgctgc taacggatcc tacccatacg atgttccaga ttacgcggcc     3900

gctccaaaaa agaaaagaaa agttgaattc ggcggcagca gcggcagcga gactcccggg     3960

acctcagagt ccgccacacc cgaaagtatg gacaagaagt atagcatcgg cctggccatc     4020

ggcacaaact ccgtgggctg ggccgtgatc accgacgagt acaaggtgcc aagcaagaag     4080

tttaaggtgc tgggcaacac cgatagacac tccatcaaga agaatctgat cggcgccctg     4140

ctgttcgact ctggcgagac agccgaggcc acacggctga agagaaccgc ccggagaagg     4200

tatacacgcc ggaagaatag gatctgctac ctgcaggaga tcttcagcaa cgagatggcc     4260

aaggtggacg attctttctt tcaccgcctg gaggagagct tcctggtgga ggaggataag     4320

aagcacgagc ggcaccctat ctttggcaac atcgtggacg aggtggccta tcacgagaag     4380

tacccaacaa tctatcacct gaggaagaag ctggtggact ccaccgataa ggccgacctg     4440

cgcctgatct atctggccct ggcccacatg atcaagttcc ggggccactt tctgatcgag     4500

ggcgatctga acccagacaa tagcgatgtg gacaagctgt tcatccagct ggtgcagacc     4560

tacaatcagc tgtttgagga gaaccccatc aatgcctctg gagtggacgc aaaggcaatc     4620

ctgagcgcca gactgtccaa gtctagaagg ctggagaacc tgatcgccca gctgccaggc     4680

gagaagaaga acggcctgtt tggcaatctg atcgccctgt ccctgggcct gacacccaac     4740

ttcaagtcta attttgatct ggccgaggac gccaagctgc agctgtccaa ggacacctat     4800

gacgatgacc tggataacct gctggcccag atcggcgatc agtacgccga cctgttcctg     4860

gccgccaaga atctgtctga cgccatcctg ctgagcgata tcctgcgcgt gaacaccgag     4920

atcacaaagg cccccctgag cgcctccatg atcaagagat atgacgagca ccaccaggat     4980

ctgaccctgc tgaaggccct ggtgaggcag cagctgcctg agaagtacaa ggagatcttc     5040

tttgatcaga gcaagaatgg atacgcagga tatatcgacg gaggagcatc ccaggaggag     5100

ttctacaagt ttatcaagcc tatcctggag aagatggacg gcacagagga gctgctggtg     5160

aagctgaatc gggaggacct gctgaggaag cagcgcacct ttgataacgg cagcatccct     5220

caccagatcc acctgggaga gctgcacgca atcctgcgcc ggcaggagga cttctaccca     5280

tttctgaagg ataaccggga gaagatcgag aagatcctga cattcagaat cccctactat     5340

gtgggacctc tggcccgggg caatagcaga tttgcctgga tgacccgcaa gtccgaggag     5400

acaatcacac cctggaactt cgaggaggtg gtggataagg gcgcctctgc ccagagcttc     5460

atcgagcgga tgaccaattt tgacaagaac ctgcctaatg agaaggtgct gccaaagcac     5520

tctctgctgt acgagtattt caccgtgtat aacgagctga caaaggtgaa gtacgtgacc     5580

gagggcatga gaaagcctgc cttcctgagc ggcgagcaga agaaggccat cgtggacctg     5640

ctgtttaaga ccaataggaa ggtgacagtg aagcagctga aggaggacta tttcaagaag     5700

atcgagtgtt ttgattctgt ggagatcagc ggcgtggagg acaggtttaa cgcctccctg     5760

ggcacctacc acgatctgct gaagatcatc aaggataagg acttcctgga caacgaggag     5820

aatgaggata tcctggagga catcgtgctg accctgacac tgtttgagga tagggagatg     5880

atcgaggagc gcctgaagac atatgcccac ctgttcgatg acaaagtgat gaagcagctg     5940

aagagaaggc gctacaccgg atggggccgg ctgagcagaa agctgatcaa tggcatccgc     6000

gacaagcagt ctggcaagac aatcctggac tttctgaaga gcgatggctt cgccaaccgg     6060

aacttcatgc agctgatcca cgatgactcc ctgaccttca aggaggatat ccagaaggca     6120

caggtgtctg gacagggcga cagcctgcac gagcacatcg ccaacctggc cggctctcct     6180

gccatcaaga agggcatcct gcagaccgtg aaggtggtgg acgagctggt gaaagtgatg     6240

ggcaggcaca agccagagaa catcgtgatc gagatggccc gcgagaatca gaccacacag     6300

aagggccaga agaactcccg ggagagaatg aagagaatcg aggagggcat caaggagctg     6360

ggctctcaga tcctgaagga gcaccccgtg gagaacacac agctgcagaa tgagaagctg     6420

tatctgtact atctgcagaa tggccgggat atgtacgtgg accaggagct ggatatcaac     6480

agactgtctg attatgacgt ggatcacatc gtgccacagt ccttcctgaa ggatgactct     6540

atcgacaata aggtgctgac caggagcgac aaggcccgcg gcaagtccga taatgtgccc     6600

tctgaggagg tggtgaagaa gatgaagaac tactggaggc agctgctgaa tgccaagctg     6660

atcacacaga ggaagtttga taacctgacc aaggcagaga ggggaggact gtccgagctg     6720

gacaaggccg gcttcatcaa gcggcagctg gtggagacaa gacagatcac aaagcacgtg     6780

gcccagatcc tggattctag aatgaacaca aagtacgatg agaatgacaa gctgatcagg     6840

gaggtgaaag tgatcaccct gaagtccaag ctggtgtctg actttaggaa ggatttccag     6900

ttttataagg tgcgcgagat caacaattat caccacgccc acgacgccta cctgaacgcc     6960

gtggtgggca cagccctgat caagaagtac cctaagctgg agtccgagtt cgtgtacggc     7020

gactataagg tgtacgatgt gcgcaagatg atcgccaagt ctgagcagga gatcggcaag     7080

gccaccgcca agtatttctt ttacagcaac atcatgaatt tctttaagac cgagatcaca     7140

ctggccaatg gcgagatcag gaagcgccca ctgatcgaga caaacggcga gacaggcgag     7200

atcgtgtggg acaagggcag ggattttgcc accgtgcgca aggtgctgag catgccccaa     7260

gtgaatatcg tgaagaagac cgaggtgcag acaggcggct tctccaagga gtctatcctg     7320

cctaagcgga actccgataa gctgatcgcc agaaagaagg actgggaccc caagaagtat     7380

ggcggcttcg acagccctac agtggcctac tccgtgctgg tggtggccaa ggtggagaag     7440

ggcaagagca agaagctgaa gtccgtgaag gagctgctgg gcatcaccat catggagcgc     7500

agctccttcg agaagaatcc tatcgacttt ctggaggcca agggctataa ggaggtgaag     7560

aaggacctga tcatcaagct gccaaagtac tctctgtttg agctggagaa cggaaggaag     7620

agaatgctgg caagcgccgg agagctgcag aagggcaatg agctggccct gccctccaag     7680

tacgtgaact tcctgtatct ggcctcccac tacgagaagc tgaagggctc tcctgaggat     7740

aacgagcaga agcagctgtt tgtggagcag cacaagcact atctggacga gatcatcgag     7800

cagatcagcg agttctccaa gagagtgatc ctggccgacg ccaatctgga taaggtgctg     7860

tccgcctaca acaagcaccg ggataagcca atcagagagc aggccgagaa tatcatccac     7920

ctgtttaccc tgacaaacct gggagcacca gcagccttca agtattttga caccacaatc     7980

gacaggaagc ggtacaccag cacaaaggag gtgctggacg ccacactgat ccaccagtcc     8040

atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga ttga           8094


<210>  62
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer A

<400>  62
ctcaggtagt ggttgtcggg c                                                 21


<210>  63
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer B

<400>  63
ggacagtggg aatctcgttc                                                   20


<210>  64
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer C

<400>  64
tgggagtctc ggcatgat                                                     18


<210>  65
<211>  67
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of Fig. 2 - Band 1 - Non-spliced - using Primer A

<400>  65
ggggtgttct gctggtagtg gtcggcgagg tgagtccagg agatgtttca gccatgttgt       60

ctttatt                                                                 67


<210>  66
<211>  932
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of Fig. 2 - Band 1 - Non-spliced - using Primer B

<400>  66
agatgacgag gcatttggct acttgaggcg agtcaccact cgctttccgg attaatgtgt       60

ccgtcacggg gacgacatcc gagtgcagcg caagatttgt aatcatgccg agactcccag      120

ctgtcccccg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc      180

tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg      240

tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt      300

cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa      360

acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca      420

gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc      480

agcggccggg aaggggcggt gcgggaggcg gggtgtgggg ctgtagtgtg ggccctgttc      540

ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc      600

ctcgttgacc gaatcaccga cctctctccc caggcaagtt tgtacaaaaa agcaggctgc      660

caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct      720

ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac      780

ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc      840

caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat      900

gaagcagcac gacttcttca agtccgccat gc                                    932


<210>  67
<211>  898
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of Fig. 2 - Band 2 - Spliced - using Primer A

<400>  67
tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc       60

ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt      120

ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga      180

tgcccttcag ctcgatgcgg ttaaccaggg tgtcgccctc gaacttcacc tcggcgcggg      240

tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca      300

tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca      360

cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc      420

agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc      480

tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga      540

acagctcctc gcccttgctc accatggtgg cagcctgctt ttttgtacaa acttgcctgg      600

ggagagaggt cggtgattcg gtcaacgagg gagccgactg ccgacgtgcg ctccggaggc      660

ttgcagaatg cggaacaccg cgcgggcagg aacagggccc acactacagc cccacacccc      720

gcctcccgca ccgccccttc ccggccgctg ctctcggcgc gccctgctga gcagccgcta      780

ttggccacag cccatcgcgg tcggcgcgct gccattgctc cctggcgctg tccgtctgcg      840

agggtactag tgagacgtgc ggcttccgtt tgtcacgtcc ggcacgccgc gaaccgca        898


<210>  68
<211>  971
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of Fig. 2 - Band 2 - Spliced - using Primer B


<220>
<221>  misc_feature
<222>  (737)..(737)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (788)..(788)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (868)..(868)
<223>  n is a, c, g, or t

<220>
<221>  misc_feature
<222>  (941)..(941)
<223>  n is a, c, g, or t

<400>  68
ttagatgacg aggcatttgg ctacttgagg cgagtcacca ctcgctttcc ggattaatgt       60

gtccgtcacg gggacgacat ccgagtgcag cgcaagattt gtaatcatgc cgagactccc      120

agctgtcccc cgggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct      180

gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca      240

cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc      300

ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac      360

aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg      420

cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga      480

gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggctggtagt gtgggccctg      540

ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc      600

tccctcgttg accgaatcac cgacctctct ccccaggcaa gtttgtacaa aaaagcaggc      660

tgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga      720

gctggacggc gacgtanacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc      780

cacctacngc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg      840

gcccaccctc gtgaccaccc tgacctangg cgtgcagtgc ttcagccgct accccgacca      900

catgaagcag cacgacttct tcaagtccgc catgcccgaa nctacgtcca ggagcgcacc      960

atcttcttca a                                                           971


<210>  69
<211>  118
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Sequence of Fig. 2 - Band 2 - Spliced - using Primer C

<400>  69
gtcgtcccgt gacggacaca ttaatccgga aagcgagtgg tgactcgcct caagtagcca       60

aatgcctcgt catctaatta gtgacgcgca tgaatggatg aacgagattc ccactgtc        118


<210>  70
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Example 4 - Forward Primer

<400>  70
tgctcaggta gtggttgtcg                                                   20


<210>  71
<211>  3188
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  R2OI_EGFP_reporter RNA sequence

<400>  71
gcgggtgttg acgcgatgtg atttctgccc agtgctctga atgtcaaagt gaagaaattc       60

aatgaagcgc gggtaaacgg cgggagtaac tatgactctc ttaaggcgca caggggacac      120

agagcctgcc caagtaccgc tcccgaggga gcgggaaacg ggggggtgac tatcccctgg      180

ggtccggcga gagcgctggt ctacggacca ggggtggctg tgggcaggct gctcctcagg      240

ccagttgatt agttacgcat gggctgtacc tccacgtggt cccgctggta acgacttgtc      300

ggctaaatca gcccgcccac catctgggat atggttgacc gtctaacccc agtactcagg      360

tcacaaacaa aatgggaaca gatacagtgt atgtcggcca ggactaccct tctggcttat      420

caaaacgggt accagcacgg ttagtggcgg gaccgatgct gcgagagcga agctgtcacg      480

cccatgtgtt tagggctgga cacatgtgga actggcgaac cagccttccg agcgggcgct      540

gggaccagcc cgctttggag aagtctcggg tcctaacccg gtcggtggcg acggccaccg      600

accccgaaat tacctcttac ccaggaaagt ccgtatcgac aagtacgcag gttcaggagg      660

aggactggtg tagccgggag agcgggtgga tctcgccagg acttgctcct gaagaaccct      720

cggtggtgtc cgaaattaca gcctccatgg tagcgacaat gagggtagca accgaggagg      780

tcgtgtaaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg      840

ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa      900

acaagttgtt tttacttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg      960

aactccagca ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg     1020

gtgctcaggt agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc     1080

tggtagtggt cggcgaggtg agtccaggag atgtttcagc actgttgcct ttagtctcga     1140

ggcaacttag acaactgagt attgatctga gcacagcagg gtgtgagctg tttgaagata     1200

ctggggttgg gagtgaagaa actgcagagg actaactggg ctgagaccca gtggcaatgt     1260

tttagggcct aaggagtgcc tctgaaaatc tagatggaca actttgactt tgagaaaaga     1320

gaggtggaaa tgaggaaaat gacttttctt tattagattt cggtagaaag aactttcacc     1380

tttcccctat ttttgttatt cgttttaaaa catctatctg gaggcaggac aagtatggtc     1440

gttaaaaaga tgcaggcaga aggcatatat tggctcagtc aaagtgggga actttggtgg     1500

ccaaacatac attgctaagg ctattcctat atcagctgga cacatataaa atgctgctaa     1560

tgcttcatta caaacttata tcctttaatt ccagatgggg gcaaagtatg tccaggggtg     1620

aggaacaatt gaaacatttg ggctggagta gattttgaaa gtcagctctg tgtgtgtgtg     1680

tgtgtgtgcg cgcgcgtgtg tgtgtgtgtg tgtcagcgtg tgtttctttt aacgtcttca     1740

gcctacaaca tacagggttc atggtgggaa gaagatagca agatttaaat tatggccagt     1800

gactagtgct gcaagaagaa caactacctg catttaatgg gaaagcaaaa tctcaggctt     1860

tgagggaagt taacataggc ttgattctgg gttgaagctg ggtgtgtagt tatctggagg     1920

ccaggctgga gctctcagct cactatgggt tcatctttat tgtctccttt catctcaaca     1980

gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct     2040

tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc     2100

ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg     2160

tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga     2220

tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca     2280

tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg     2340

gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg     2400

tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca     2460

gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg     2520

cagcctgctt ttttgtacaa acttgcctgg ggagagaggt cggtgattcg gtcaacgagg     2580

gagccgactg ccgacgtgcg ctccggaggc ttgcagaatg cggaacaccg cgcgggcagg     2640

aacagggccc acactaccgc cccacacccc gcctcccgca ccgccccttc ccggccgctg     2700

ctctcggcgc gccctgctga gcagccgcta ttggccacag cccatcgcgg tcggcgcgct     2760

gccattgctc cctggcgctg tccgtctgcg agggtactag tgagacgtgc ggcttccgtt     2820

tgtcacgtcc ggcacgccgc gaaccgcaag gaaccttccc gacttagggg cggagcagga     2880

agcgtcgccg gggggcccac aagggtagcg gcgaagatcc gggtgacgct gcgaacggac     2940

gtgaagaatg tgcgagaccc agggtcggcg ccgctgcgtt tcccggaacc acgcccagag     3000

cagccgcgtc cctgcgcaaa cccagggctg ccttggaaaa ggcgcaaccc gggggacagc     3060

tgggagtctc ggcatgatta caaatcttgc gctgcactcg gatgtcgtcc ccgtgacgga     3120

cacattaatc cggaaagcga gtggtgactc gcctcaagta gccaaatgcc tcgtcatcta     3180

attagtga                                                              3188


<210>  72
<211>  2265
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  EGFP cassette sequence

<400>  72
taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta       60

tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag      120

ttgtttttac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc      180

cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct      240

caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta      300

gtggtcggcg aggtgagtcc aggagatgtt tcagcactgt tgcctttagt ctcgaggcaa      360

cttagacaac tgagtattga tctgagcaca gcagggtgtg agctgtttga agatactggg      420

gttgggagtg aagaaactgc agaggactaa ctgggctgag acccagtggc aatgttttag      480

ggcctaagga gtgcctctga aaatctagat ggacaacttt gactttgaga aaagagaggt      540

ggaaatgagg aaaatgactt ttctttatta gatttcggta gaaagaactt tcacctttcc      600

cctatttttg ttattcgttt taaaacatct atctggaggc aggacaagta tggtcgttaa      660

aaagatgcag gcagaaggca tatattggct cagtcaaagt ggggaacttt ggtggccaaa      720

catacattgc taaggctatt cctatatcag ctggacacat ataaaatgct gctaatgctt      780

cattacaaac ttatatcctt taattccaga tgggggcaaa gtatgtccag gggtgaggaa      840

caattgaaac atttgggctg gagtagattt tgaaagtcag ctctgtgtgt gtgtgtgtgt      900

gtgcgcgcgc gtgtgtgtgt gtgtgtgtca gcgtgtgttt cttttaacgt cttcagccta      960

caacatacag ggttcatggt gggaagaaga tagcaagatt taaattatgg ccagtgacta     1020

gtgctgcaag aagaacaact acctgcattt aatgggaaag caaaatctca ggctttgagg     1080

gaagttaaca taggcttgat tctgggttga agctgggtgt gtagttatct ggaggccagg     1140

ctggagctct cagctcacta tgggttcatc tttattgtct cctttcatct caacagctgc     1200

acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc     1260

ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg     1320

atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg     1380

ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg     1440

cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg     1500

tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag     1560

ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca     1620

tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg     1680

accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcagcc     1740

tgcttttttg tacaaacttg cctggggaga gaggtcggtg attcggtcaa cgagggagcc     1800

gactgccgac gtgcgctccg gaggcttgca gaatgcggaa caccgcgcgg gcaggaacag     1860

ggcccacact accgccccac accccgcctc ccgcaccgcc ccttcccggc cgctgctctc     1920

ggcgcgccct gctgagcagc cgctattggc cacagcccat cgcggtcggc gcgctgccat     1980

tgctccctgg cgctgtccgt ctgcgagggt actagtgaga cgtgcggctt ccgtttgtca     2040

cgtccggcac gccgcgaacc gcaaggaacc ttcccgactt aggggcggag caggaagcgt     2100

cgccgggggg cccacaaggg tagcggcgaa gatccgggtg acgctgcgaa cggacgtgaa     2160

gaatgtgcga gacccagggt cggcgccgct gcgtttcccg gaaccacgcc cagagcagcc     2220

gcgtccctgc gcaaacccag ggctgccttg gaaaaggcgc aaccc                     2265


