               SEQUENCE LISTING

<110> Biospirál-2006 Fejlesztő és Tanácsadó Kft.

<120> VARIANT CAS9

<130> P132002

<140> HUPCT/2021
<141> 2021-02-25

<150> PCT/HU2020/050063
<151> 2020-12-23
<150> HU2000083
<151> 2020-03-06
<150> HU2000072
<151> 2020-02-26
<150> HU2000068
<151> 2020-02-25

<160> 584

<170> BiSSAP 1.3.6

<210> 1
<211> 1423
<212> PRT
<213> Artificial Sequence


<220> 
<221> PEPTIDE
<222> 1..23
<223> 3X FLAG® Peptide

<220> 
<223> Mutant (variant) Cas9 protein of the invention

<220> 
<221> SIGNAL
<222> 24..40
<223> Nuclear localization signal

<220> 
<221> SITE
<222> 1042
<223> Corresponds to Lys1003 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1043
<223> Corresponds to Leu1004 in the sequence with N-terminal methionine

<220> 
<221> SITE
<222> 1046
<223> Corresponds to Glu1007 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1052
<223> Corresponds to Tyr1013 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1053
<223> Corresponds to Lys1014 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1056
<223> Corresponds to Asp1017 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SIGNAL
<222> 1408..1423
<223> Nuclear localization signal

<400> 1
Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      
Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          
Gly Ile His Gly Val Pro Ala Ala Asp Lys Lys Tyr Ser Ile Gly Leu 
        35                  40                  45              
Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
    50                  55                  60                  
Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
65                  70                  75                  80  
Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
                85                  90                  95      
Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
            100                 105                 110         
Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
        115                 120                 125             
Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
    130                 135                 140                 
Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
145                 150                 155                 160 
Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
                165                 170                 175     
Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
            180                 185                 190         
Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
        195                 200                 205             
Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
    210                 215                 220                 
Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
225                 230                 235                 240 
Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
                245                 250                 255     
Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
            260                 265                 270         
Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
        275                 280                 285             
Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
    290                 295                 300                 
Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
305                 310                 315                 320 
Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
                325                 330                 335     
Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
            340                 345                 350         
Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
        355                 360                 365             
Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
    370                 375                 380                 
Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
385                 390                 395                 400 
Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
                405                 410                 415     
Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
            420                 425                 430         
Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
        435                 440                 445             
Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
    450                 455                 460                 
Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
465                 470                 475                 480 
Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
                485                 490                 495     
Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
            500                 505                 510         
Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
        515                 520                 525             
Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu 
    530                 535                 540                 
Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
545                 550                 555                 560 
Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
                565                 570                 575     
Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
            580                 585                 590         
Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe 
        595                 600                 605             
Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
    610                 615                 620                 
Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile 
625                 630                 635                 640 
Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu 
                645                 650                 655     
Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu 
            660                 665                 670         
Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys 
        675                 680                 685             
Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys 
    690                 695                 700                 
Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp 
705                 710                 715                 720 
Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile 
                725                 730                 735     
His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val 
            740                 745                 750         
Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly 
        755                 760                 765             
Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp 
    770                 775                 780                 
Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile 
785                 790                 795                 800 
Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser 
                805                 810                 815     
Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser 
            820                 825                 830         
Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu 
        835                 840                 845             
Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp 
    850                 855                 860                 
Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile 
865                 870                 875                 880 
Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu 
                885                 890                 895     
Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu 
            900                 905                 910         
Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala 
        915                 920                 925             
Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg 
    930                 935                 940                 
Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu 
945                 950                 955                 960 
Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser 
                965                 970                 975     
Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val 
            980                 985                 990         
Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp 
        995                 1000                1005            
Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His 
    1010                1015                1020                
Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr 
1025                1030                1035                1040
Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp 
                1045                1050                1055    
Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr 
            1060                1065                1070        
Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu 
        1075                1080                1085            
Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr 
    1090                1095                1100                
Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala 
1105                1110                1115                1120
Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys 
                1125                1130                1135    
Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys 
            1140                1145                1150        
Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys 
        1155                1160                1165            
Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val 
    1170                1175                1180                
Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys 
1185                1190                1195                1200
Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn 
                1205                1210                1215    
Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp 
            1220                1225                1230        
Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly 
        1235                1240                1245            
Arg Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu 
    1250                1255                1260                
Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His 
1265                1270                1275                1280
Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu 
                1285                1290                1295    
Phe Val Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile 
            1300                1305                1310        
Ser Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys 
        1315                1320                1325            
Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln 
    1330                1335                1340                
Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro 
1345                1350                1355                1360
Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr 
                1365                1370                1375    
Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr 
            1380                1385                1390        
Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Lys 
        1395                1400                1405            
Arg Pro Ala Ala Thr Lys Lys Ala Gly Gln Ala Lys Lys Lys Lys 
    1410                1415                1420            

<210> 2
<211> 1367
<212> PRT
<213> Streptococcus pyogenes serotype M1


<220> 
<221> SITE
<222> 1002
<223> Corresponds to Lys1003 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1003
<223> Corresponds to Leu1004 in the sequence with N-terminal methionine

<220> 
<221> SITE
<222> 1006
<223> Corresponds to Glu1007 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1012
<223> Corresponds to Tyr1013 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1013
<223> Corresponds to Lys1014 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1016
<223> Corresponds to Asp1017 in the wt Cas9 sequence with N-terminal
      methionine

<400> 2
Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val Gly 
1               5                   10                  15      
Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys 
            20                  25                  30          
Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly 
        35                  40                  45              
Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys 
    50                  55                  60                  
Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr 
65                  70                  75                  80  
Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe 
                85                  90                  95      
Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His 
            100                 105                 110         
Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His 
        115                 120                 125             
Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser 
    130                 135                 140                 
Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met 
145                 150                 155                 160 
Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp 
                165                 170                 175     
Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn 
            180                 185                 190         
Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys 
        195                 200                 205             
Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu 
    210                 215                 220                 
Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu 
225                 230                 235                 240 
Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp 
                245                 250                 255     
Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp 
            260                 265                 270         
Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu 
        275                 280                 285             
Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile 
    290                 295                 300                 
Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met 
305                 310                 315                 320 
Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala 
                325                 330                 335     
Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp 
            340                 345                 350         
Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln 
        355                 360                 365             
Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
    370                 375                 380                 
Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys 
385                 390                 395                 400 
Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly 
                405                 410                 415     
Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu 
            420                 425                 430         
Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
        435                 440                 445             
Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met 
    450                 455                 460                 
Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val 
465                 470                 475                 480 
Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
                485                 490                 495     
Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu 
            500                 505                 510         
Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
        515                 520                 525             
Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys 
    530                 535                 540                 
Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
545                 550                 555                 560 
Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
                565                 570                 575     
Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
            580                 585                 590         
Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
        595                 600                 605             
Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
    610                 615                 620                 
Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
625                 630                 635                 640 
Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr 
                645                 650                 655     
Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys 
            660                 665                 670         
Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala 
        675                 680                 685             
Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys 
    690                 695                 700                 
Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His 
705                 710                 715                 720 
Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile 
                725                 730                 735     
Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg 
            740                 745                 750         
His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr 
        755                 760                 765             
Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu 
    770                 775                 780                 
Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val 
785                 790                 795                 800 
Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln 
                805                 810                 815     
Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu 
            820                 825                 830         
Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys Asp 
        835                 840                 845             
Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly 
    850                 855                 860                 
Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn 
865                 870                 875                 880 
Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe 
                885                 890                 895     
Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys 
            900                 905                 910         
Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys 
        915                 920                 925             
His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu 
    930                 935                 940                 
Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys 
945                 950                 955                 960 
Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu 
                965                 970                 975     
Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val 
            980                 985                 990         
Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val 
        995                 1000                1005            
Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser 
    1010                1015                1020                
Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn 
1025                1030                1035                1040
Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile 
                1045                1050                1055    
Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val 
            1060                1065                1070        
Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met 
        1075                1080                1085            
Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe 
    1090                1095                1100                
Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala 
1105                1110                1115                1120
Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro 
                1125                1130                1135    
Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys 
            1140                1145                1150        
Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met 
        1155                1160                1165            
Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys 
    1170                1175                1180                
Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr 
1185                1190                1195                1200
Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala 
                1205                1210                1215    
Gly 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 Pro 
        1235                1240                1245            
Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr 
    1250                1255                1260                
Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile 
1265                1270                1275                1280
Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys His 
                1285                1290                1295    
Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His Leu Phe 
            1300                1305                1310        
Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr 
        1315                1320                1325            
Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala 
    1330                1335                1340                
Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp 
1345                1350                1355                1360
Leu Ser Gln Leu Gly Gly Asp 
                1365        

<210> 3
<211> 1370
<212> PRT
<213> Streptococcus pyogenes serotype M4


<220> 
<221> SITE
<222> 1001
<223> Corresponds to Lys1003 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1002
<223> Corresponds to Leu1004 in the sequence with N-terminal methionine

<220> 
<221> SITE
<222> 1005
<223> Corresponds to Glu1007 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1011
<223> Corresponds to Tyr1013 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1012
<223> Corresponds to Lys1014 in the wt Cas9 sequence with N-terminal
      methionine

<220> 
<221> SITE
<222> 1015
<223> Corresponds to Asp1017 in the wt Cas9 sequence with N-terminal
      methionine

<400> 3
Asp Lys Lys Tyr Ser Ile Gly Leu Asp Ile Gly Thr Asn Ser Val Gly 
1               5                   10                  15      
Trp Ala Val Ile Thr Asp Asp Tyr Lys Val Pro Ser Lys Lys Phe Lys 
            20                  25                  30          
Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly 
        35                  40                  45              
Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys 
    50                  55                  60                  
Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr 
65                  70                  75                  80  
Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe 
                85                  90                  95      
Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His 
            100                 105                 110         
Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His 
        115                 120                 125             
Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Ala Asp Ser 
    130                 135                 140                 
Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met 
145                 150                 155                 160 
Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp 
                165                 170                 175     
Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn 
            180                 185                 190         
Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Arg Val Asp Ala Lys 
        195                 200                 205             
Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu 
    210                 215                 220                 
Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu 
225                 230                 235                 240 
Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp 
                245                 250                 255     
Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp 
            260                 265                 270         
Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu 
        275                 280                 285             
Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Thr Leu Leu Ser Asp Ile 
    290                 295                 300                 
Leu Arg Val Asn Ser Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met 
305                 310                 315                 320 
Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala 
                325                 330                 335     
Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp 
            340                 345                 350         
Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln 
        355                 360                 365             
Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly 
    370                 375                 380                 
Thr Glu Glu Leu Leu Ala Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys 
385                 390                 395                 400 
Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly 
                405                 410                 415     
Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu 
            420                 425                 430         
Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro 
        435                 440                 445             
Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met 
    450                 455                 460                 
Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val 
465                 470                 475                 480 
Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn 
                485                 490                 495     
Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu 
            500                 505                 510         
Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr 
        515                 520                 525             
Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys 
    530                 535                 540                 
Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val 
545                 550                 555                 560 
Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser 
                565                 570                 575     
Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr 
            580                 585                 590         
Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn 
        595                 600                 605             
Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu 
    610                 615                 620                 
Phe Glu Asp Lys Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His 
625                 630                 635                 640 
Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg His Tyr Thr 
                645                 650                 655     
Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys 
            660                 665                 670         
Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala 
        675                 680                 685             
Asn Arg Asn Phe Ile Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys 
    690                 695                 700                 
Glu Ala Ile Gln Lys Ala Gln Val Ser Gly Gln Gly His Ser Leu His 
705                 710                 715                 720 
Glu Gln Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile 
                725                 730                 735     
Leu Gln Ser Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly His 
            740                 745                 750         
Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr 
        755                 760                 765             
Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu 
    770                 775                 780                 
Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu 
785                 790                 795                 800 
Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn 
                805                 810                 815     
Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser 
            820                 825                 830         
Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Ile Lys Asp Asp 
        835                 840                 845             
Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys 
    850                 855                 860                 
Ser Asp Asp Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr 
865                 870                 875                 880 
Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp 
                885                 890                 895     
Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala 
            900                 905                 910         
Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His 
        915                 920                 925             
Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn 
    930                 935                 940                 
Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu 
945                 950                 955                 960 
Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile 
                965                 970                 975     
Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly 
            980                 985                 990         
Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr 
        995                 1000                1005            
Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu 
    1010                1015                1020                
Gln Glu Ile Gly Lys Ala Thr Ala Lys Arg Phe Phe Tyr Ser Asn Ile 
1025                1030                1035                1040
Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg 
                1045                1050                1055    
Lys Arg Pro Leu Ile Glu Thr Asn Glu Glu Thr Gly Glu Ile Val Trp 
            1060                1065                1070        
Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro 
        1075                1080                1085            
Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly Ala Leu Thr 
    1090                1095                1100                
Asn Glu Ser Ile Tyr Ala Arg Gly Ser Phe Asp Lys Leu Ile Ser Arg 
1105                1110                1115                1120
Lys His Arg Phe Glu Ser Ser Lys Tyr Gly Gly Phe Gly Ser Pro Thr 
                1125                1130                1135    
Val Thr Tyr Ser Val Leu Val Val Ala Lys Ser Lys Val Gln Asp Gly 
            1140                1145                1150        
Lys Val Lys Lys Ile Lys Thr Gly Lys Glu Leu Ile Gly Ile Thr Leu 
        1155                1160                1165            
Leu Asp Lys Leu Val Phe Glu Lys Asn Pro Leu Lys Phe Ile Glu Asp 
    1170                1175                1180                
Lys Gly Tyr Gly Asn Val Gln Ile Asp Lys Cys Ile Lys Leu Pro Lys 
1185                1190                1195                1200
Tyr Ser Leu Phe Glu Phe Glu Asn Gly Thr Arg Arg Met Leu Ala Ser 
                1205                1210                1215    
Val Met Ala Asn Asn Asn Ser Arg Gly Asp Leu Gln Lys Ala Asn Glu 
            1220                1225                1230        
Met Phe Leu Pro Ala Lys Leu Val Thr Leu Leu Tyr His Ala His Lys 
        1235                1240                1245            
Ile Glu Ser Ser Lys Glu Leu Glu His Glu Ala Tyr Ile Leu Asp His 
    1250                1255                1260                
Tyr Asn Asp Leu Tyr Gln Leu Leu Ser Tyr Ile Glu Arg Phe Ala Ser 
1265                1270                1275                1280
Leu Tyr Val Asp Val Glu Lys Asn Ile Ser Lys Val Lys Glu Leu Phe 
                1285                1290                1295    
Ser Asn Ile Glu Ser Tyr Ser Ile Ser Glu Ile Cys Ser Ser Val Ile 
            1300                1305                1310        
Asn Leu Leu Thr Leu Thr Ala Ser Gly Ala Pro Ala Asp Phe Lys Phe 
        1315                1320                1325            
Leu Gly Thr Thr Ile Pro Arg Lys Arg Tyr Gly Ser Pro Gln Ser Ile 
    1330                1335                1340                
Leu Ser Ser Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr 
1345                1350                1355                1360
Arg Ile Asp Leu Ser Gln Leu Gly Ser Asp 
                1365                1370

<210> 4
<211> 15
<212> PRT
<213> Streptococcus pyogenes


<220> 
<223> Wild type loop sequence

<400> 4
Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp 
1               5                   10                  15  

<210> 5
<211> 15
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B1 mutant loop sequence

<400> 5
Lys Leu Glu Ser Gly Phe Val Tyr Gly Asp Gly Lys Val Tyr Asp 
1               5                   10                  15  

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


<220> 
<223> HF-B2 mutant loop sequence

<400> 6
Lys Gly Lys Val Tyr Asp 
1               5       

<210> 7
<211> 15
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B3 mutant loop sequence

<220> 
<221> SITE
<222> 2
<223> Xaa is deletion

<220> 
<221> SITE
<222> 3
<223> Xaa is deletion

<220> 
<221> SITE
<222> 4
<223> Xaa is deletion

<220> 
<221> SITE
<222> 5
<223> Xaa is deletion

<220> 
<221> SITE
<222> 6
<223> Xaa is deletion

<220> 
<221> SITE
<222> 7
<223> Xaa is deletion

<220> 
<221> SITE
<222> 8
<223> Xaa is deletion

<220> 
<221> SITE
<222> 9
<223> Xaa is deletion

<220> 
<221> SITE
<222> 10
<223> Xaa is deletion

<220> 
<221> SITE
<222> 11
<223> Xaa is deletion

<220> 
<221> SITE
<222> 12
<223> Xaa is deletion

<220> 
<221> SITE
<222> 13
<223> Xaa is deletion

<220> 
<221> SITE
<222> 14
<223> Xaa is deletion

<400> 7
Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp 
1               5                   10                  15  

<210> 8
<211> 15
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B4 mutant loop sequence

<220> 
<221> SITE
<222> 2
<223> Xaa is deletion

<220> 
<221> SITE
<222> 3
<223> Xaa is deletion

<220> 
<221> SITE
<222> 4
<223> Xaa is deletion

<220> 
<221> SITE
<222> 5
<223> Xaa is deletion

<220> 
<221> SITE
<222> 6
<223> Xaa is deletion

<220> 
<221> SITE
<222> 8
<223> Xaa is deletion

<220> 
<221> SITE
<222> 9
<223> Xaa is deletion

<220> 
<221> SITE
<222> 10
<223> Xaa is deletion

<220> 
<221> SITE
<222> 11
<223> Xaa is deletion

<220> 
<221> SITE
<222> 12
<223> Xaa is deletion

<220> 
<221> SITE
<222> 13
<223> Xaa is deletion

<220> 
<221> SITE
<222> 14
<223> Xaa is deletion

<400> 8
Lys Xaa Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp 
1               5                   10                  15  

<210> 9
<211> 4
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B5 mutant loop sequence

<400> 9
Lys Gly Lys Asp 
1               

<210> 10
<211> 6
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B6 mutant loop sequence

<400> 10
Lys Leu Lys Val Tyr Asp 
1               5       

<210> 11
<211> 7
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B7 mutant loop sequence

<400> 11
Lys Leu Gly Lys Val Tyr Asp 
1               5           

<210> 12
<211> 7
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B8 mutant loop sequence

<400> 12
Lys Leu Pro Lys Val Tyr Asp 
1               5           

<210> 13
<211> 8
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B9 (B-SpCas9-HF1) mutant loop sequence

<400> 13
Lys Leu Gly Gly Lys Val Tyr Asp 
1               5               

<210> 14
<211> 8
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B10 mutant loop sequence

<400> 14
Lys Leu Gly Pro Lys Val Tyr Asp 
1               5               

<210> 15
<211> 9
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B11 mutant loop sequence

<400> 15
Lys Leu Gly Gly Gly Lys Val Tyr Asp 
1               5                   

<210> 16
<211> 10
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B12 mutant loop sequence

<400> 16
Lys Leu Gly Gly Gly Gly Lys Val Tyr Asp 
1               5                   10  

<210> 17
<211> 7
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B13 mutant loop sequence

<400> 17
Lys Leu Gly Gly Val Tyr Asp 
1               5           

<210> 18
<211> 6
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B14 mutant loop sequence

<400> 18
Lys Leu Gly Gly Tyr Asp 
1               5       

<210> 19
<211> 5
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B15 mutant loop sequence

<400> 19
Lys Leu Gly Gly Asp 
1               5   

<210> 20
<211> 8
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B16 mutant loop sequence

<400> 20
Lys Leu Ser Ser Lys Val Tyr Asp 
1               5               

<210> 21
<211> 5
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B17 mutant loop sequence

<400> 21
Lys Leu Ser Ser Asp 
1               5   

<210> 22
<211> 8
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B18 mutant loop sequence

<400> 22
Lys Leu Glu Ser Lys Val Tyr Asp 
1               5               

<210> 23
<211> 9
<212> PRT
<213> Artificial Sequence


<220> 
<223> HF-B19 mutant loop sequence

<400> 23
Lys Leu Glu Ser Gly Lys Val Tyr Asp 
1               5                   

<210> 24
<211> 4101
<212> DNA
<213> Streptococcus pyogenes


<220> 
<223> Nucleotide sequence coding wild type cas9 protein

<400> 24
gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc       60

accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac      120

agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc      180

acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat      240

ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg      300

gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac      360

atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa      420

ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg      480

atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg      540

gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc      600

aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg      660

ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg      720

attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat      780

gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag      840

atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg      900

ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg      960

atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag     1020

cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc     1080

tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa     1140

aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag     1200

cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc     1260

attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag     1320

aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga     1380

ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg     1440

gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac     1500

ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat     1560

aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc     1620

ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg     1680

aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc     1740

ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc     1800

aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg     1860

accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac     1920

ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg     1980

ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat     2040

ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc     2100

ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac     2160

gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg     2220

aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc     2280

gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg     2340

aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg     2400

gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat     2460

atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc     2520

gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac     2580

aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac     2640

tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc     2700

aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg     2760

gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact     2820

aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag     2880

ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac     2940

caccacgccc acgacgcgta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac     3000

cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt acggaagatg     3060

atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac     3120

atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct     3180

ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc     3240

accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag     3300

acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc     3360

agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat     3420

tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa     3480

gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt     3540

ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac     3600

tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag     3660

aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac     3720

tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc     3840

ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc     3900

atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct     3960

gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag     4020

gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac     4080

ctgtctcagc tgggaggcga c                                               4101


<210> 25
<211> 53
<212> DNA
<213> Artificial Sequence


<220> 
<223> i5-FANCF2-ONtarget-FWD PCR primer

<400> 25
tcgtcggcag cgtcagatgt gtataagaga cagggtgctg acgtaggtag tgc            53


<210> 26
<211> 55
<212> DNA
<213> Artificial Sequence


<220> 
<223> i7-FANCF2-ONtarget-REV PCR primer

<400> 26
gtctcgtggg ctcggagatg tgtataagag acaggtcaca gtatgtctct ggcgt          55


<210> 27
<211> 53
<212> DNA
<213> Artificial Sequence


<220> 
<223> i5-FANCF2-ONtarget-FWD2 PCR primer

<400> 27
tcgtcggcag cgtcagatgt gtataagaga caggggccgg gaaagagttg ctg            53


<210> 28
<211> 56
<212> DNA
<213> Artificial Sequence


<220> 
<223> i7-FANCF2-ONtarget-REV2 PCR primer

<400> 28
gtctcgtggg ctcggagatg tgtataagag acaggcccta catctgctct ccctcc         56


<210> 29
<211> 53
<212> DNA
<213> Artificial Sequence


<220> 
<223> i5-VEGFA3-ONtarget-FWD2 PCR primer

<400> 29
tcgtcggcag cgtcagatgt gtataagaga cagtccagat ggcacattgt cag            53


<210> 30
<211> 54
<212> DNA
<213> Artificial Sequence


<220> 
<223> i7-FVEGFA3-ONtarget-REV2 PCR primer

<400> 30
gtctcgtggg ctcggagatg tgtataagag acagagggag caggaaagtg aggt           54


<210> 31
<211> 53
<212> DNA
<213> Artificial Sequence


<220> 
<223> i5-VEGFA3-ONtarget-REV2 PCR primer

<400> 31
tcgtcggcag cgtcagatgt gtataagaga cagagggagc aggaaagtga ggt            53


<210> 32
<211> 54
<212> DNA
<213> Artificial Sequence


<220> 
<223> i7-FVEGFA3-ONtarget-FWD2 PCR primer

<400> 32
gtctcgtggg ctcggagatg tgtataagag acagtccaga tggcacattg tcag           54


<210> 33
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site51 primer

<400> 33
aaaaaataat acgactcact atagcgatgc ccttcagctc gatg                      44


<210> 34
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site52 primer

<400> 34
aaaaaataat acgactcact atagcaacat cctggggcac aagc                      44


<210> 35
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site53 primer

<400> 35
aaaaaataat acgactcact atagcagctc gatgcggttc acca                      44


<210> 36
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site54 primer

<400> 36
aaaaaataat acgactcact atagcaagga ggacggcaac atcc                      44


<210> 37
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site55 primer

<400> 37
aaaaaataat acgactcact atagtcagct cgatgcggtt cacc                      44


<210> 38
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site56 primer

<400> 38
aaaaaataat acgactcact atagaaggag gacggcaaca tcct                      44


<210> 39
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site57 primer

<400> 39
aaaaaataat acgactcact atagaggagg acggcaacat cctg                      44


<210> 40
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site58 primer

<400> 40
aaaaaataat acgactcact atagcatgcc cgaaggctac gtcc                      44


<210> 41
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site59 primer

<400> 41
aaaaaataat acgactcact atagcgtgct gcttcatgtg gtcg                      44


<210> 42
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site51 primer

<400> 42
aaaaaataat acgactcact ataggcgatg cccttcagct cgatg                     45


<210> 43
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site52 primer

<400> 43
aaaaaataat acgactcact ataggcaaca tcctggggca caagc                     45


<210> 44
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site53 primer

<400> 44
aaaaaataat acgactcact ataggcagct cgatgcggtt cacca                     45


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


<220> 
<223> T7-+2G-Site54 primer

<400> 45
aaaaaataat acgactcact ataggcaagg aggacggcaa catcc                     45


<210> 46
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site55 primer

<400> 46
aaaaaataat acgactcact ataggtcagc tcgatgcggt tcacc                     45


<210> 47
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site56 primer

<400> 47
aaaaaataat acgactcact ataggaagga ggacggcaac atcct                     45


<210> 48
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site57 primer

<400> 48
aaaaaataat acgactcact ataggaggag gacggcaaca tcctg                     45


<210> 49
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site58 primer

<400> 49
aaaaaataat acgactcact ataggcatgc ccgaaggcta cgtcc                     45


<210> 50
<211> 45
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+2G-Site59 primer

<400> 50
aaaaaataat acgactcact ataggcgtgc tgcttcatgt ggtcg                     45


<210> 51
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site-1 primer

<400> 51
aaaaaataat acgactcact atagggcacg ggcagcttgc cgg                       43


<210> 52
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site3 primer

<400> 52
aaaaaataat acgactcact ataggtggtg cagatgaact tca                       43


<210> 53
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site4 primer

<400> 53
aaaaaataat acgactcact ataggagcgc accatcttct tca                       43


<210> 54
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site16 primer

<400> 54
aaaaaataat acgactcact ataggcgagg gcgatgccac cta                       43


<210> 55
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site21 primer

<400> 55
aaaaaataat acgactcact ataggcatcg acttcaagga gga                       43


<210> 56
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site22 primer

<400> 56
aaaaaataat acgactcact ataggtgttc tgctggtagt ggt                       43


<210> 57
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site31 primer

<400> 57
aaaaaataat acgactcact atagggcgag gagctgttca ccg                       43


<210> 58
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site33 primer

<400> 58
aaaaaataat acgactcact ataggtgccc atcctggtcg agc                       43


<210> 59
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site37 primer

<400> 59
aaaaaataat acgactcact ataggtcagg gtggtcacga ggg                       43


<210> 60
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site46 primer

<400> 60
aaaaaataat acgactcact ataggttgtc gggcagcagc acg                       43


<210> 61
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site48 primer

<400> 61
aaaaaataat acgactcact ataggtgctc aggtagtggt tgt                       43


<210> 62
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site1 primer

<400> 62
aaaaaataat acgactcact ataggggcac gggcagcttg ccgg                      44


<210> 63
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site3 primer

<400> 63
aaaaaataat acgactcact atagggtggt gcagatgaac ttca                      44


<210> 64
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site4 primer

<400> 64
aaaaaataat acgactcact atagggagcg caccatcttc ttca                      44


<210> 65
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site16 primer

<400> 65
aaaaaataat acgactcact atagggcgag ggcgatgcca ccta                      44


<210> 66
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site21 primer

<400> 66
aaaaaataat acgactcact atagggcatc gacttcaagg agga                      44


<210> 67
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site22 primer

<400> 67
aaaaaataat acgactcact atagggtgtt ctgctggtag tggt                      44


<210> 68
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site31 primer

<400> 68
aaaaaataat acgactcact ataggggcga ggagctgttc accg                      44


<210> 69
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site33 primer

<400> 69
aaaaaataat acgactcact atagggtgcc catcctggtc gagc                      44


<210> 70
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site37 primer

<400> 70
aaaaaataat acgactcact atagggtcag ggtggtcacg aggg                      44


<210> 71
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site46 primer

<400> 71
aaaaaataat acgactcact atagggttgt cgggcagcag cacg                      44


<210> 72
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site48 primer

<400> 72
aaaaaataat acgactcact atagggtgct caggtagtgg ttgt                      44


<210> 73
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site5 primer

<400> 73
aaaaaataat acgactcact atagtcgccc tcgaacttca cct                       43


<210> 74
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site13 primer

<400> 74
aaaaaataat acgactcact atagctgaag ggcatcgact tca                       43


<210> 75
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site17 primer

<400> 75
aaaaaataat acgactcact atagctgaag cactgcacgc cgt                       43


<210> 76
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site18 primer

<400> 76
aaaaaataat acgactcact atagtgaacc gcatcgagct gaa                       43


<210> 77
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site20 primer

<400> 77
aaaaaataat acgactcact atagacgtag ccttcgggca tgg                       43


<210> 78
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site23 primer

<400> 78
aaaaaataat acgactcact atagtggtca cgagggtggg cca                       43


<210> 79
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site38 primer

<400> 79
aaaaaataat acgactcact atagtaggtc agggtggtca cga                       43


<210> 80
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site39 primer

<400> 80
aaaaaataat acgactcact atagcactgc acgccgtagg tca                       43


<210> 81
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site44 primer

<400> 81
aaaaaataat acgactcact atagatgccg ttcttctgct tgt                       43


<210> 82
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site45 primer

<400> 82
aaaaaataat acgactcact atagcacggg gccgtcgccg atg                       43


<210> 83
<211> 43
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-Site47 primer

<400> 83
aaaaaataat acgactcact atagtggttg tcgggcagca gca                       43


<210> 84
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site5 primer

<400> 84
aaaaaataat acgactcact ataggtcgcc ctcgaacttc acct                      44


<210> 85
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site13 primer

<400> 85
aaaaaataat acgactcact ataggctgaa gggcatcgac ttca                      44


<210> 86
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site17 primer

<400> 86
aaaaaataat acgactcact ataggctgaa gcactgcacg ccgt                      44


<210> 87
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site18 primer

<400> 87
aaaaaataat acgactcact ataggtgaac cgcatcgagc tgaa                      44


<210> 88
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site20 primer

<400> 88
aaaaaataat acgactcact ataggacgta gccttcgggc atgg                      44


<210> 89
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site23 primer

<400> 89
aaaaaataat acgactcact ataggtggtc acgagggtgg gcca                      44


<210> 90
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site38 primer

<400> 90
aaaaaataat acgactcact ataggtaggt cagggtggtc acga                      44


<210> 91
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site39 primer

<400> 91
aaaaaataat acgactcact ataggcactg cacgccgtag gtca                      44


<210> 92
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site44 primer

<400> 92
aaaaaataat acgactcact ataggatgcc gttcttctgc ttgt                      44


<210> 93
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site45 primer

<400> 93
aaaaaataat acgactcact ataggcacgg ggccgtcgcc gatg                      44


<210> 94
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site47 primer

<400> 94
aaaaaataat acgactcact ataggtggtt gtcgggcagc agca                      44


<210> 95
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-Site43 primer

<400> 95
aaaaaataat acgactcact ataggaaggg catcgacttc aagg                      44


<210> 96
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-VEGFAsite2 primer

<400> 96
aaaaaataat acgactcact ataggacccc ctccaccccg cctc                      44


<210> 97
<211> 44
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-+G-FANCFsite2 primer

<400> 97
aaaaaataat acgactcact ataggctgca gaagggattc catg                      44


<210> 98
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> T7-reverse primer

<400> 98
aaaaaagcac cgactcggtg cc                                             22


<210> 99
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 2-for primer

<400> 99
gggccgggaa agagttgctg                                                20


<210> 100
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 2-rev primer

<400> 100
gccctacatc tgctctccct cc                                             22


<210> 101
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 2-for primer

<400> 101
cgaggaagag agagacgggg tc                                             22


<210> 102
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 2-rev primer

<400> 102
ctccaatgca cccaagacag cag                                            23


<210> 103
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> HEK site 4-for primer

<400> 103
agagaagttg gagtgaaggc agag                                           24


<210> 104
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> HEK site 4-rev primer

<400> 104
gtcagacgtc caaaaccaga ctcc                                           24


<210> 105
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> ZSCAN2-for primer

<400> 105
agtgtggggt gtgtgggaag                                                20


<210> 106
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> ZSCAN2-rev primer

<400> 106
acgggacttg actcagacca ct                                             22


<210> 107
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EMX1 site 2-for primer

<400> 107
ggagcagctg gtcagagggg                                                20


<210> 108
<211> 22
<212> DNA
<213> Artificial Sequence


<220> 
<223> EMX1 site 2-rev

<400> 108
ccatagggaa gggggacact gg                                             22


<210> 109
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> DNMT1 site 4-for primer

<400> 109
ccagaatgca caaagtactg cac                                            23


<210> 110
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> DNMT1 site 4-rev

<400> 110
gccaaagccc gagagagtgc c                                              21


<210> 111
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 1 spacer sequence

<400> 111
gggcacgggc agcttgccgg                                                20


<210> 112
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 2 spacer sequence

<400> 112
gagctggacg gcgacgtaaa                                                20


<210> 113
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 3 spacer sequence

<400> 113
ggtggtgcag atgaacttca                                                20


<210> 114
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 4 spacer sequence

<400> 114
ggagcgcacc atcttcttca                                                20


<210> 115
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 5 spacer sequence

<400> 115
gtcgccctcg aacttcacct                                                20


<210> 116
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 6 spacer sequence

<400> 116
gcgcgatcac atggtcctgc                                                20


<210> 117
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 7 spacer sequence

<400> 117
gtccatgccg agagtgatcc                                                20


<210> 118
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 8 spacer sequence

<400> 118
gccgtcgtcc ttgaagaaga                                                20


<210> 119
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 9 spacer sequence

<400> 119
gaagttcgag ggcgacaccc                                                20


<210> 120
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 10 spacer sequence

<400> 120
gtctttgctc agggcggact                                                20


<210> 121
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 11 spacer sequence

<400> 121
ggtctttgct cagggcggac                                                20


<210> 122
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 12 spacer sequence

<400> 122
gtgctcaggt agtggttgtc                                                20


<210> 123
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 13 spacer sequence

<400> 123
gctgaagggc atcgacttca                                                20


<210> 124
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 14 spacer sequence

<400> 124
gtaccagcac tagcctcctg                                                20


<210> 125
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 15 spacer sequence

<400> 125
ggcatcgccc tcgccctcgc                                                20


<210> 126
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 16 spacer sequence

<400> 126
ggcgagggcg atgccaccta                                                20


<210> 127
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 17 spacer sequence

<400> 127
gctgaagcac tgcacgccgt                                                20


<210> 128
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 18 spacer sequence

<400> 128
gtgaaccgca tcgagctgaa                                                20


<210> 129
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 19

<400> 129
gtcagggtgg tcacgagggt                                                20


<210> 130
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 20 spacer sequence

<400> 130
gacgtagcct tcgggcatgg                                                20


<210> 131
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 21 spacer sequence

<400> 131
ggcatcgact tcaaggagga                                                20


<210> 132
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 22 spacer sequence

<400> 132
ggtgttctgc tggtagtggt                                                20


<210> 133
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 23 spacer sequence

<400> 133
gtggtcacga gggtgggcca                                                20


<210> 134
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 24 spacer sequence

<400> 134
gtcgtgctgc ttcatgtggt                                                20


<210> 135
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 25 spacer sequence

<400> 135
ggtccgagag tctgtagcca                                                20


<210> 136
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 26 spacer sequence

<400> 136
gctgacggtc aggagccagg                                                20


<210> 137
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 27 spacer sequence

<400> 137
ggcagagcag gctgacggtc                                                20


<210> 138
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 28 spacer sequence

<400> 138
gagcaggcag agcaggctga                                                20


<210> 139
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 29 spacer sequence

<400> 139
gaggccagag caggcagagc                                                20


<210> 140
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 30 spacer sequence

<400> 140
gctctgcctg ctctggcctc                                                20


<210> 141
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 31 spacer sequence

<400> 141
gggcgaggag ctgttcaccg                                                20


<210> 142
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 32 spacer sequence

<400> 142
gaccaggatg ggcaccaccc                                                20


<210> 143
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 33 spacer sequence

<400> 143
ggtgcccatc ctggtcgagc                                                20


<210> 144
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 34 spacer sequence

<400> 144
gccgtccagc tcgaccagga                                                20


<210> 145
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 35 spacer sequence

<400> 145
ggccacaagt tcagcgtgtc                                                20


<210> 146
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 36 spacer sequence

<400> 146
ggtggtcacg agggtgggcc                                                20


<210> 147
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 37 spacer sequence

<400> 147
ggtcagggtg gtcacgaggg                                                20


<210> 148
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 38 spacer sequence

<400> 148
gtaggtcagg gtggtcacga                                                20


<210> 149
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 39 spacer sequence

<400> 149
gcactgcacg ccgtaggtca                                                20


<210> 150
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 40 spacer sequence

<400> 150
gcttcatgtg gtcggggtag                                                20


<210> 151
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 41 spacer sequence

<400> 151
gcgctcctgg acgtagcctt                                                20


<210> 152
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 42 spacer sequence

<400> 152
ggtgaaccgc atcgagctga                                                20


<210> 153
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 43 spacer sequence

<400> 153
gaagggcatc gacttcaagg                                                20


<210> 154
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 44 spacer sequence

<400> 154
gatgccgttc ttctgcttgt                                                20


<210> 155
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 45 spacer sequence

<400> 155
gcacggggcc gtcgccgatg                                                20


<210> 156
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 46 spacer sequence

<400> 156
ggttgtcggg cagcagcacg                                                20


<210> 157
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 47 spacer sequence

<400> 157
gtggttgtcg ggcagcagca                                                20


<210> 158
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 48 spacer sequence

<400> 158
ggtgctcagg tagtggttgt                                                20


<210> 159
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 49 spacer sequence

<400> 159
gttggggtct ttgctcaggg                                                20


<210> 160
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 50 spacer sequence

<400> 160
gccgagagtg atcccggcgg                                                20


<210> 161
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 51-no 5' G spacer sequence

<400> 161
cgatgccctt cagctcgatg                                                20


<210> 162
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 52-no 5' G spacer sequence

<400> 162
caacatcctg gggcacaagc                                                20


<210> 163
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 53-no 5' G spacer sequence

<400> 163
cagctcgatg cggttcacca                                                20


<210> 164
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 54-no 5' G spacer sequence

<400> 164
caaggaggac ggcaacatcc                                                20


<210> 165
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 55-no 5' G spacer sequence

<400> 165
tcagctcgat gcggttcacc                                                20


<210> 166
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 56-no 5' G spacer sequence

<400> 166
aaggaggacg gcaacatcct                                                20


<210> 167
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 57-no 5' G spacer sequence

<400> 167
aggaggacgg caacatcctg                                                20


<210> 168
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 58-no 5' G spacer sequence

<400> 168
catgcccgaa ggctacgtcc                                                20


<210> 169
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 59-no 5' G spacer sequence

<400> 169
cgtgctgctt catgtggtcg                                                20


<210> 170
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 1B spacer sequence

<400> 170
ggggcacggg cagcttgccg g                                              21


<210> 171
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 2B spacer sequence

<400> 171
ggagctggac ggcgacgtaa a                                              21


<210> 172
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 3B spacer sequence

<400> 172
gggtggtgca gatgaacttc a                                              21


<210> 173
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 4B spacer sequence

<400> 173
gggagcgcac catcttcttc a                                              21


<210> 174
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 5B spacer sequence

<400> 174
ggtcgccctc gaacttcacc t                                              21


<210> 175
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 6B spacer sequence

<400> 175
ggcgcgatca catggtcctg c                                              21


<210> 176
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 7B spacer sequence

<400> 176
ggtccatgcc gagagtgatc c                                              21


<210> 177
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 8B spacer sequence

<400> 177
ggccgtcgtc cttgaagaag a                                              21


<210> 178
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 9B spacer sequence

<400> 178
ggaagttcga gggcgacacc c                                              21


<210> 179
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 13B spacer sequence

<400> 179
ggctgaaggg catcgacttc a                                              21


<210> 180
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 14B spacer sequence

<400> 180
ggtaccagca ctagcctcct g                                              21


<210> 181
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 15B spacer sequence

<400> 181
gggcatcgcc ctcgccctcg c                                              21


<210> 182
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 24B spacer sequence

<400> 182
ggtcgtgctg cttcatgtgg t                                              21


<210> 183
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 25B spacer sequence

<400> 183
gggtccgaga gtctgtagcc a                                              21


<210> 184
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 27B spacer sequence

<400> 184
gggcagagca ggctgacggt c                                              21


<210> 185
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 28B spacer sequence

<400> 185
ggagcaggca gagcaggctg a                                              21


<210> 186
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 29B spacer sequence

<400> 186
ggaggccaga gcaggcagag c                                              21


<210> 187
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 30B spacer sequence

<400> 187
ggctctgcct gctctggcct c                                              21


<210> 188
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 31B spacer sequence

<400> 188
ggggcgagga gctgttcacc g                                              21


<210> 189
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 32B spacer sequence

<400> 189
ggaccaggat gggcaccacc c                                              21


<210> 190
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 33B spacer sequence

<400> 190
gggtgcccat cctggtcgag c                                              21


<210> 191
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 34B spacer sequence

<400> 191
ggccgtccag ctcgaccagg a                                              21


<210> 192
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 35B spacer sequence

<400> 192
gggccacaag ttcagcgtgt c                                              21


<210> 193
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 37B spacer sequence

<400> 193
gggtcagggt ggtcacgagg g                                              21


<210> 194
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 38B spacer sequence

<400> 194
ggtaggtcag ggtggtcacg a                                              21


<210> 195
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 39B spacer sequence

<400> 195
ggcactgcac gccgtaggtc a                                              21


<210> 196
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 40B spacer sequence

<400> 196
ggcttcatgt ggtcggggta g                                              21


<210> 197
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 41B spacer sequence

<400> 197
ggcgctcctg gacgtagcct t                                              21


<210> 198
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 42B spacer sequence

<400> 198
gggtgaaccg catcgagctg a                                              21


<210> 199
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 43B spacer sequence

<400> 199
ggaagggcat cgacttcaag g                                              21


<210> 200
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 44B spacer sequence

<400> 200
ggatgccgtt cttctgcttg t                                              21


<210> 201
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 45B spacer sequence

<400> 201
ggcacggggc cgtcgccgat g                                              21


<210> 202
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 46B spacer sequence

<400> 202
gggttgtcgg gcagcagcac g                                              21


<210> 203
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 47B spacer sequence

<400> 203
ggtggttgtc gggcagcagc a                                              21


<210> 204
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 49B spacer sequence

<400> 204
ggttggggtc tttgctcagg g                                              21


<210> 205
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 50B spacer sequence

<400> 205
ggccgagagt gatcccggcg g                                              21


<210> 206
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 10C spacer sequence

<400> 206
ggtctttgct cagggcggac t                                              21


<210> 207
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 11C spacer sequence

<400> 207
gggtctttgc tcagggcgga c                                              21


<210> 208
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 12C spacer sequence

<400> 208
ggtgctcagg tagtggttgt c                                              21


<210> 209
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 16C spacer sequence

<400> 209
gggcgagggc gatgccacct a                                              21


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


<220> 
<223> EGFP site 17C spacer sequence

<400> 210
ggctgaagca ctgcacgccg t                                              21


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


<220> 
<223> EGFP site 18C spacer sequence

<400> 211
ggtgaaccgc atcgagctga a                                              21


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


<220> 
<223> EGFP site 19C spacer sequence

<400> 212
ggtcagggtg gtcacgaggg t                                              21


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


<220> 
<223> EGFP site 20C spacer sequence

<400> 213
ggacgtagcc ttcgggcatg g                                              21


<210> 214
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 21C spacer sequence

<400> 214
gggcatcgac ttcaaggagg a                                              21


<210> 215
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 22C spacer sequence

<400> 215
gggtgttctg ctggtagtgg t                                              21


<210> 216
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 23C spacer sequence

<400> 216
ggtggtcacg agggtgggcc a                                              21


<210> 217
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 26C spacer sequence

<400> 217
ggctgacggt caggagccag g                                              21


<210> 218
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 36C spacer sequence

<400> 218
gggtggtcac gagggtgggc c                                              21


<210> 219
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 48C spacer sequence

<400> 219
gggtgctcag gtagtggttg t                                              21


<210> 220
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 1 target sequence with PAM

<400> 220
gggcacgggc agcttgccgg tgg                                            23


<210> 221
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 2 target sequence with PAM

<400> 221
gagctggacg gcgacgtaaa cgg                                            23


<210> 222
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 3 target sequence with PAM

<400> 222
ggtggtgcag atgaacttca ggg                                            23


<210> 223
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 4 target sequence with PAM

<400> 223
ggagcgcacc atcttcttca agg                                            23


<210> 224
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 5 target sequence with PAM

<400> 224
gtcgccctcg aacttcacct cgg                                            23


<210> 225
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 6 target sequence with PAM

<400> 225
gcgcgatcac atggtcctgc tgg                                            23


<210> 226
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 7 target sequence with PAM

<400> 226
gtccatgccg agagtgatcc cgg                                            23


<210> 227
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 8 target sequence with PAM

<400> 227
gccgtcgtcc ttgaagaaga tgg                                            23


<210> 228
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 9 target sequence with PAM

<400> 228
gaagttcgag ggcgacaccc tgg                                            23


<210> 229
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 10 target sequence with PAM

<400> 229
gtctttgctc agggcggact ggg                                            23


<210> 230
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 11 target sequence with PAM

<400> 230
ggtctttgct cagggcggac tgg                                            23


<210> 231
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 12 target sequence with PAM

<400> 231
gtgctcaggt agtggttgtc ggg                                            23


<210> 232
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 13 target sequence with PAM

<400> 232
gctgaagggc atcgacttca agg                                            23


<210> 233
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 14 target sequence with PAM

<400> 233
gtaccagcac tagcctcctg agg                                            23


<210> 234
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 15 target sequence with PAM

<400> 234
ggcatcgccc tcgccctcgc cgg                                            23


<210> 235
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 16 target sequence with PAM

<400> 235
ggcgagggcg atgccaccta cgg                                            23


<210> 236
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 17 target sequence with PAM

<400> 236
gctgaagcac tgcacgccgt agg                                            23


<210> 237
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 18 target sequence with PAM

<400> 237
gtgaaccgca tcgagctgaa ggg                                            23


<210> 238
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 19 target sequence with PAM

<400> 238
gtcagggtgg tcacgagggt ggg                                            23


<210> 239
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 20 target sequence with PAM

<400> 239
gacgtagcct tcgggcatgg cgg                                            23


<210> 240
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 21 target sequence with PAM

<400> 240
ggcatcgact tcaaggagga cgg                                            23


<210> 241
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 22 target sequence with PAM

<400> 241
ggtgttctgc tggtagtggt cgg                                            23


<210> 242
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 23 target sequence with PAM

<400> 242
gtggtcacga gggtgggcca ggg                                            23


<210> 243
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 24 target sequence with PAM

<400> 243
gtcgtgctgc ttcatgtggt cgg                                            23


<210> 244
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 25 target sequence with PAM

<400> 244
ggtccgagag tctgtagcca tgg                                            23


<210> 245
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 26 target sequence with PAM

<400> 245
gctgacggtc aggagccagg agg                                            23


<210> 246
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 27 target sequence with PAM

<400> 246
ggcagagcag gctgacggtc agg                                            23


<210> 247
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 28 target sequence with PAM

<400> 247
gagcaggcag agcaggctga cgg                                            23


<210> 248
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 29 target sequence with PAM

<400> 248
gaggccagag caggcagagc agg                                            23


<210> 249
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 30 target sequence with PAM

<400> 249
gctctgcctg ctctggcctc agg                                            23


<210> 250
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 31 target sequence with PAM

<400> 250
gggcgaggag ctgttcaccg ggg                                            23


<210> 251
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 32 target sequence with PAM

<400> 251
gaccaggatg ggcaccaccc cgg                                            23


<210> 252
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 33 target sequence with PAM

<400> 252
ggtgcccatc ctggtcgagc tgg                                            23


<210> 253
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 34 target sequence with PAM

<400> 253
gccgtccagc tcgaccagga tgg                                            23


<210> 254
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 35 target sequence with PAM

<400> 254
ggccacaagt tcagcgtgtc cgg                                            23


<210> 255
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 36 target sequence with PAM

<400> 255
ggtggtcacg agggtgggcc agg                                            23


<210> 256
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 37 target sequence with PAM

<400> 256
ggtcagggtg gtcacgaggg tgg                                            23


<210> 257
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 38 target sequence with PAM

<400> 257
gtaggtcagg gtggtcacga ggg                                            23


<210> 258
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 39 target sequence with PAM

<400> 258
gcactgcacg ccgtaggtca ggg                                            23


<210> 259
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 40 target sequence with PAM

<400> 259
gcttcatgtg gtcggggtag cgg                                            23


<210> 260
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 41 target sequence with PAM

<400> 260
gcgctcctgg acgtagcctt cgg                                            23


<210> 261
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 42 target sequence with PAM

<400> 261
ggtgaaccgc atcgagctga agg                                            23


<210> 262
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 43 target sequence with PAM

<400> 262
gaagggcatc gacttcaagg agg                                            23


<210> 263
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 44 target sequence with PAM

<400> 263
gatgccgttc ttctgcttgt cgg                                            23


<210> 264
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 45 target sequence with PAM

<400> 264
gcacggggcc gtcgccgatg ggg                                            23


<210> 265
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 46 target sequence with PAM

<400> 265
ggttgtcggg cagcagcacg ggg                                            23


<210> 266
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 47 target sequence with PAM

<400> 266
gtggttgtcg ggcagcagca cgg                                            23


<210> 267
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 48 target sequence with PAM

<400> 267
ggtgctcagg tagtggttgt cgg                                            23


<210> 268
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 49 target sequence with PAM

<400> 268
gttggggtct ttgctcaggg cgg                                            23


<210> 269
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 50 target sequence with PAM

<400> 269
gccgagagtg atcccggcgg cgg                                            23


<210> 270
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 51-no 5' G target sequence with PAM

<400> 270
cgatgccctt cagctcgatg cgg                                            23


<210> 271
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 52-no 5' G target sequence with PAM

<400> 271
caacatcctg gggcacaagc tgg                                            23


<210> 272
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 53-no 5' G target sequence with PAM

<400> 272
cagctcgatg cggttcacca ggg                                            23


<210> 273
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 54-no 5' G target sequence with PAM

<400> 273
caaggaggac ggcaacatcc tgg                                            23


<210> 274
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 55-no 5' G target sequence with PAM

<400> 274
tcagctcgat gcggttcacc agg                                            23


<210> 275
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 56-no 5' G target sequence with PAM

<400> 275
aaggaggacg gcaacatcct ggg                                            23


<210> 276
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 57-no 5' G target sequence with PAM

<400> 276
aggaggacgg caacatcctg ggg                                            23


<210> 277
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 58-no 5' G target sequence with PAM

<400> 277
catgcccgaa ggctacgtcc agg                                            23


<210> 278
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 59-no 5' G target sequence with PAM

<400> 278
cgtgctgctt catgtggtcg ggg                                            23


<210> 279
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 1B target sequence with PAM

<400> 279
gggcacgggc agcttgccgg tgg                                            23


<210> 280
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 2B target sequence with PAM

<400> 280
gagctggacg gcgacgtaaa cgg                                            23


<210> 281
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 3B target sequence with PAM

<400> 281
ggtggtgcag atgaacttca ggg                                            23


<210> 282
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 4B target sequence with PAM

<400> 282
ggagcgcacc atcttcttca agg                                            23


<210> 283
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 5B target sequence with PAM

<400> 283
gtcgccctcg aacttcacct cgg                                            23


<210> 284
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 6B target sequence with PAM

<400> 284
gcgcgatcac atggtcctgc tgg                                            23


<210> 285
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 7B target sequence with PAM

<400> 285
gtccatgccg agagtgatcc cgg                                            23


<210> 286
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 8B target sequence with PAM

<400> 286
gccgtcgtcc ttgaagaaga tgg                                            23


<210> 287
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 9B target sequence with PAM

<400> 287
gaagttcgag ggcgacaccc tgg                                            23


<210> 288
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 13B target sequence with PAM

<400> 288
gctgaagggc atcgacttca agg                                            23


<210> 289
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 14B target sequence with PAM

<400> 289
gtaccagcac tagcctcctg agg                                            23


<210> 290
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 15B target sequence with PAM

<400> 290
ggcatcgccc tcgccctcgc cgg                                            23


<210> 291
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 24B target sequence with PAM

<400> 291
gtcgtgctgc ttcatgtggt cgg                                            23


<210> 292
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 25B target sequence with PAM

<400> 292
ggtccgagag tctgtagcca tgg                                            23


<210> 293
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 27B target sequence with PAM

<400> 293
ggcagagcag gctgacggtc agg                                            23


<210> 294
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 28B target sequence with PAM

<400> 294
gagcaggcag agcaggctga cgg                                            23


<210> 295
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 29B target sequence with PAM

<400> 295
gaggccagag caggcagagc agg                                            23


<210> 296
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 30B target sequence with PAM

<400> 296
gctctgcctg ctctggcctc agg                                            23


<210> 297
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 31B target sequence with PAM

<400> 297
gggcgaggag ctgttcaccg ggg                                            23


<210> 298
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 32B target sequence with PAM

<400> 298
gaccaggatg ggcaccaccc cgg                                            23


<210> 299
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 33B target sequence with PAM

<400> 299
ggtgcccatc ctggtcgagc tgg                                            23


<210> 300
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 34B target sequence with PAM

<400> 300
gccgtccagc tcgaccagga tgg                                            23


<210> 301
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 35B target sequence with PAM

<400> 301
ggccacaagt tcagcgtgtc cgg                                            23


<210> 302
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 37B target sequence with PAM

<400> 302
ggtcagggtg gtcacgaggg tgg                                            23


<210> 303
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 38B target sequence with PAM

<400> 303
gtaggtcagg gtggtcacga ggg                                            23


<210> 304
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 39B target sequence with PAM

<400> 304
gcactgcacg ccgtaggtca ggg                                            23


<210> 305
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 40B target sequence with PAM

<400> 305
gcttcatgtg gtcggggtag cgg                                            23


<210> 306
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 41B target sequence with PAM

<400> 306
gcgctcctgg acgtagcctt cgg                                            23


<210> 307
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 42B target sequence with PAM

<400> 307
ggtgaaccgc atcgagctga agg                                            23


<210> 308
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 43B target sequence with PAM

<400> 308
gaagggcatc gacttcaagg agg                                            23


<210> 309
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 44B target sequence with PAM

<400> 309
gatgccgttc ttctgcttgt cgg                                            23


<210> 310
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 45B target sequence with PAM

<400> 310
gcacggggcc gtcgccgatg ggg                                            23


<210> 311
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 46B target sequence with PAM

<400> 311
ggttgtcggg cagcagcacg ggg                                            23


<210> 312
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 47B target sequence with PAM

<400> 312
gtggttgtcg ggcagcagca cgg                                            23


<210> 313
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 49B target sequence with PAM

<400> 313
gttggggtct ttgctcaggg cgg                                            23


<210> 314
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 50B target sequence with PAM

<400> 314
gccgagagtg atcccggcgg cgg                                            23


<210> 315
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 10C target sequence with PAM

<400> 315
ggtctttgct cagggcggac tggg                                           24


<210> 316
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 11C target sequence with PAM

<400> 316
ggtctttgct cagggcggac tgg                                            23


<210> 317
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 12C target sequence with PAM

<400> 317
ggtgctcagg tagtggttgt cggg                                           24


<210> 318
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 16C target sequence with PAM

<400> 318
gggcgagggc gatgccacct acgg                                           24


<210> 319
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 17C target sequence with PAM

<400> 319
ggctgaagca ctgcacgccg tagg                                           24


<210> 320
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 18C target sequence with PAM

<400> 320
ggtgaaccgc atcgagctga aggg                                           24


<210> 321
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 19C target sequence with PAM

<400> 321
ggtcagggtg gtcacgaggg tggg                                           24


<210> 322
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 20C target sequence with PAM

<400> 322
ggacgtagcc ttcgggcatg gcgg                                           24


<210> 323
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 21C target sequence with PAM

<400> 323
gggcatcgac ttcaaggagg acgg                                           24


<210> 324
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 22C target sequence with PAM

<400> 324
gggtgttctg ctggtagtgg tcgg                                           24


<210> 325
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 23C target sequence with PAM

<400> 325
ggtggtcacg agggtgggcc aggg                                           24


<210> 326
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 26C target sequence with PAM

<400> 326
gctgacggtc aggagccagg agg                                            23


<210> 327
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 36C target sequence with PAM

<400> 327
ggtggtcacg agggtgggcc agg                                            23


<210> 328
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 48C target sequence with PAM

<400> 328
ggtgctcagg tagtggttgt cgg                                            23


<210> 329
<400> 329
000

<210> 330
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 51 target sequence with PAM

<400> 330
cgatgccctt cagctcgatg cgg                                            23


<210> 331
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 52 target sequence with PAM

<400> 331
caacatcctg gggcacaagc tgg                                            23


<210> 332
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 53 target sequence with PAM

<400> 332
cagctcgatg cggttcacca ggg                                            23


<210> 333
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 55 target sequence with PAM

<400> 333
tcagctcgat gcggttcacc agg                                            23


<210> 334
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 59 target sequence with PAM

<400> 334
cgtgctgctt catgtggtcg ggg                                            23


<210> 335
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 60 target sequence with PAM

<400> 335
ccgtccagct cgaccaggat ggg                                            23


<210> 336
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 61 target sequence with PAM

<400> 336
caactacaag acccgcgccg agg                                            23


<210> 337
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 62 target sequence with PAM

<400> 337
aagggcgagg agctgttcac cgg                                            23


<210> 338
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 63 target sequence with PAM

<400> 338
aagttcagcg tgtccggcga ggg                                            23


<210> 339
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 64 target sequence with PAM

<400> 339
agcgtgtccg gcgagggcga ggg                                            23


<210> 340
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 65 target sequence with PAM

<400> 340
acgagggtgg gccagggcac ggg                                            23


<210> 341
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 66 target sequence

<400> 341
agaagtcgtg ctgcttcatg tgg                                            23


<210> 342
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 67 target sequence with PAM

<400> 342
accatcttct tcaaggacga cgg                                            23


<210> 343
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 68 target sequence with PAM

<400> 343
aaggaggacg gcaacatcct ggg                                            23


<210> 344
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 69 target sequence with PAM

<400> 344
tcagctcgat gcggttcacc agg                                            23


<210> 345
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 70 target sequence with PAM

<400> 345
tcgtgctgct tcatgtggtc ggg                                            23


<210> 346
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 71 target sequence with PAM

<400> 346
ttcaagtccg ccatgcccga agg                                            23


<210> 347
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 72 target sequence with PAM

<400> 347
tgaagaagat ggtgcgctcc tgg                                            23


<210> 348
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 73 target sequence with PAM

<400> 348
tgtactccag cttgtgcccc agg                                            23


<210> 349
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 74 target sequence with PAM

<400> 349
tgccgtcctc gatgttgtgg cgg                                            23


<210> 350
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 75 target sequence with PAM

<400> 350
ccgcgccgag gtgaagttcg agg                                            23


<210> 351
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 51 21G-sgRNA (spacer sequence)

<400> 351
gcgatgccct tcagctcgat g                                              21


<210> 352
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 52 21G-sgRNA (spacer sequence)

<400> 352
gcaacatcct ggggcacaag c                                              21


<210> 353
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 53 21G-sgRNA (spacer sequence)

<400> 353
gcagctcgat gcggttcacc a                                              21


<210> 354
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 55 21G-sgRNA (spacer sequence)

<400> 354
gtcagctcga tgcggttcac c                                              21


<210> 355
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 59 21G-sgRNA (spacer sequence)

<400> 355
gcgtgctgct tcatgtggtc g                                              21


<210> 356
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 60 21G-sgRNA (spacer sequence)

<400> 356
gccgtccagc tcgaccagga t                                              21


<210> 357
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 61 21G-sgRNA (spacer sequence)

<400> 357
gcaactacaa gacccgcgcc g                                              21


<210> 358
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 62 21G-sgRNA (spacer sequence)

<400> 358
gaagggcgag gagctgttca c                                              21


<210> 359
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 63 21G-sgRNA (spacer sequence)

<400> 359
gaagttcagc gtgtccggcg a                                              21


<210> 360
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 64 21G-sgRNA (spacer sequence)

<400> 360
gagcgtgtcc ggcgagggcg a                                              21


<210> 361
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 65 21G-sgRNA (spacer sequence)

<400> 361
gacgagggtg ggccagggca c                                              21


<210> 362
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 66 21G-sgRNA (spacer sequence)

<400> 362
gagaagtcgt gctgcttcat g                                              21


<210> 363
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 67 21G-sgRNA (spacer sequence)

<400> 363
gaccatcttc ttcaaggacg a                                              21


<210> 364
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 68 21G-sgRNA (spacer sequence)

<400> 364
gaaggaggac ggcaacatcc t                                              21


<210> 365
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 69 21G-sgRNA (spacer sequence)

<400> 365
gtcagctcga tgcggttcac c                                              21


<210> 366
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 70 21G-sgRNA (spacer sequence)

<400> 366
gtcgtgctgc ttcatgtggt c                                              21


<210> 367
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 71 21G-sgRNA (spacer sequence)

<400> 367
gttcaagtcc gccatgcccg a                                              21


<210> 368
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 72 21G-sgRNA (spacer sequence)

<400> 368
gtgaagaaga tggtgcgctc c                                              21


<210> 369
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 73 21G-sgRNA (spacer sequence)

<400> 369
gtgtactcca gcttgtgccc c                                              21


<210> 370
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 74 21G-sgRNA (spacer sequence)

<400> 370
gtgccgtcct cgatgttgtg g                                              21


<210> 371
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 75 21G-sgRNA (spacer sequence)

<400> 371
gccgcgccga ggtgaagttc g                                              21


<210> 372
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 51 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 372
gcatcgctga tgagtccgtg aggacgaaac gagtaagctc gtccgatgcc cttcagctcg     60

atg                                                                   63


<210> 373
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 52 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 373
atgttgctga tgagtccgtg aggacgaaac gagtaagctc gtccaacatc ctggggcaca     60

agc                                                                   63


<210> 374
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 53 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 374
gagctgctga tgagtccgtg aggacgaaac gagtaagctc gtccagctcg atgcggttca     60

cca                                                                   63


<210> 375
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 55 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 375
agctgactga tgagtccgtg aggacgaaac gagtaagctc gtctcagctc gatgcggttc     60

acc                                                                   63


<210> 376
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 59 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 376
agcacgctga tgagtccgtg aggacgaaac gagtaagctc gtccgtgctg cttcatgtgg     60

tcg                                                                   63


<210> 377
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 60 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 377
ggacggctga tgagtccgtg aggacgaaac gagtaagctc gtcccgtcca gctcgaccag     60

gat                                                                   63


<210> 378
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 61 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 378
tagttgctga tgagtccgtg aggacgaaac gagtaagctc gtccaactac aagacccgcg     60

ccg                                                                   63


<210> 379
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 62 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 379
gcccttctga tgagtccgtg aggacgaaac gagtaagctc gtcaagggcg aggagctgtt     60

cac                                                                   63


<210> 380
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 63 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 380
gaacttctga tgagtccgtg aggacgaaac gagtaagctc gtcaagttca gcgtgtccgg     60

cga                                                                   63


<210> 381
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 64 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 381
cacgctctga tgagtccgtg aggacgaaac gagtaagctc gtcagcgtgt ccggcgaggg     60

cga                                                                   63


<210> 382
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 65 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 382
cctcgtctga tgagtccgtg aggacgaaac gagtaagctc gtcacgaggg tgggccaggg     60

cac                                                                   63


<210> 383
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 66 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 383
acttctctga tgagtccgtg aggacgaaac gagtaagctc gtcagaagtc gtgctgcttc     60

atg                                                                   63


<210> 384
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 67 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 384
gatggtctga tgagtccgtg aggacgaaac gagtaagctc gtcaccatct tcttcaagga     60

cga                                                                   63


<210> 385
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 68 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 385
ctccttctga tgagtccgtg aggacgaaac gagtaagctc gtcaaggagg acggcaacat     60

cct                                                                   63


<210> 386
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 69 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 386
agctgactga tgagtccgtg aggacgaaac gagtaagctc gtctcagctc gatgcggttc     60

acc                                                                   63


<210> 387
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 70 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 387
gcacgactga tgagtccgtg aggacgaaac gagtaagctc gtctcgtgct gcttcatgtg     60

gtc                                                                   63


<210> 388
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 71 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 388
cttgaactga tgagtccgtg aggacgaaac gagtaagctc gtcttcaagt ccgccatgcc     60

cga                                                                   63


<210> 389
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 72 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 389
tcttcactga tgagtccgtg aggacgaaac gagtaagctc gtctgaagaa gatggtgcgc     60

tcc                                                                   63


<210> 390
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 73 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 390
agtacactga tgagtccgtg aggacgaaac gagtaagctc gtctgtactc cagcttgtgc     60

ccc                                                                   63


<210> 391
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 74 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 391
acggcactga tgagtccgtg aggacgaaac gagtaagctc gtctgccgtc ctcgatgttg     60

tgg                                                                   63


<210> 392
<211> 63
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 75 ribosyme flanked sgRNA (ribosyme-spacer sequence)

<400> 392
gcgcggctga tgagtccgtg aggacgaaac gagtaagctc gtcccgcgcc gaggtgaagt     60

tcg                                                                   63


<210> 393
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 51 tRNA flanked sgRNA (spacer sequence)

<400> 393
cgatgccctt cagctcgatg                                                20


<210> 394
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 52 tRNA flanked sgRNA (spacer sequence)

<400> 394
caacatcctg gggcacaagc                                                20


<210> 395
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 53 tRNA flanked sgRNA (spacer sequence)

<400> 395
cagctcgatg cggttcacca                                                20


<210> 396
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 55 tRNA flanked sgRNA (spacer sequence)

<400> 396
tcagctcgat gcggttcacc                                                20


<210> 397
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 59 tRNA flanked sgRNA (spacer sequence)

<400> 397
cgtgctgctt catgtggtcg                                                20


<210> 398
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 60 tRNA flanked sgRNA (spacer sequence)

<400> 398
ccgtccagct cgaccaggat                                                20


<210> 399
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 61 tRNA flanked sgRNA (spacer sequence)

<400> 399
caactacaag acccgcgccg                                                20


<210> 400
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 62 tRNA flanked sgRNA (spacer sequence)

<400> 400
aagggcgagg agctgttcac                                                20


<210> 401
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 63 tRNA flanked sgRNA (spacer sequence)

<400> 401
aagttcagcg tgtccggcga                                                20


<210> 402
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 64 tRNA flanked sgRNA (spacer sequence)

<400> 402
agcgtgtccg gcgagggcga                                                20


<210> 403
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 65 tRNA flanked sgRNA (spacer sequence)

<400> 403
acgagggtgg gccagggcac                                                20


<210> 404
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 66 tRNA flanked sgRNA (spacer sequence)

<400> 404
agaagtcgtg ctgcttcatg                                                20


<210> 405
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 67 tRNA flanked sgRNA (spacer sequence)

<400> 405
accatcttct tcaaggacga                                                20


<210> 406
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 68 tRNA flanked sgRNA (spacer sequence)

<400> 406
aaggaggacg gcaacatcct                                                20


<210> 407
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 69 tRNA flanked sgRNA (spacer sequence)

<400> 407
tcagctcgat gcggttcacc                                                20


<210> 408
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 70 tRNA flanked sgRNA (spacer sequence)

<400> 408
tcgtgctgct tcatgtggtc                                                20


<210> 409
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 71 tRNA flanked sgRNA (spacer sequence)

<400> 409
ttcaagtccg ccatgcccga                                                20


<210> 410
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 72 tRNA flanked sgRNA (spacer sequence)

<400> 410
tgaagaagat ggtgcgctcc                                                20


<210> 411
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 73 tRNA flanked sgRNA (spacer sequence)

<400> 411
tgtactccag cttgtgcccc                                                20


<210> 412
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 74 tRNA flanked sgRNA (spacer sequence)

<400> 412
tgccgtcctc gatgttgtgg                                                20


<210> 413
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 75 tRNA flanked sgRNA (spacer sequence)

<400> 413
ccgcgccgag gtgaagttcg                                                20


<210> 414
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 1 spacer sequence

<400> 414
gcgatccagg tgctgcagaa                                                20


<210> 415
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 2 spacer sequence

<400> 415
gctgcagaag ggattccatg                                                20


<210> 416
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 3 spacer sequence

<400> 416
gtagggcctt cgcgcacctc a                                              21


<210> 417
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 4 spacer sequence

<400> 417
gccaaggtga aagcggaagt a                                              21


<210> 418
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 5 spacer sequence

<400> 418
gtccaaggtg aaagcggaag t                                              21


<210> 419
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 6 spacer sequence

<400> 419
gcgccgtctc caaggtgaaa g                                              21


<210> 420
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 7 spacer sequence

<400> 420
gcctgctctc tctgcgcctg c                                              21


<210> 421
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 8 spacer sequence

<400> 421
gccagcaggc gcagagagag c                                              21


<210> 422
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 9 spacer sequence

<400> 422
gcaggacgtc acagtgaccg a                                              21


<210> 423
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 10 spacer sequence

<400> 423
gttagcgaac ttccaggccc t                                              21


<210> 424
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 11 spacer sequence

<400> 424
gcgtcacagt gaccgagggc c                                              21


<210> 425
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 12 spacer sequence

<400> 425
gtccgggatt agcgaacttc c                                              21


<210> 426
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 1 spacer sequence

<400> 426
gcttcatgta cagagagccc a                                              21


<210> 427
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 3 spacer sequence

<400> 427
gaacagctac atatttggga c                                              21


<210> 428
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 4 spacer sequence

<400> 428
gtcccaaata tgtagctgtt                                                20


<210> 429
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 5 spacer sequence

<400> 429
gcaaatatgt agctgtttgg g                                              21


<210> 430
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 6 spacer sequence

<400> 430
gtttgggagg tcagaaatag g                                              21


<210> 431
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 7 spacer sequence

<400> 431
gggtgggggg agtttgctcc                                                20


<210> 432
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 8 spacer sequence

<400> 432
gctttgctag gaatattgaa g                                              21


<210> 433
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 9 spacer sequence

<400> 433
ggcaggggaa ggcggagagc                                                20


<210> 434
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 10 spacer sequence

<400> 434
gtctccccac ccgtccctgt c                                              21


<210> 435
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 11 spacer sequence

<400> 435
gagagccgga cagggacggg                                                20


<210> 436
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 12 spacer sequence

<400> 436
ggacagggac gggtggggag                                                20


<210> 437
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 1B spacer sequence

<400> 437
gttctgccca gtaggatgaa c                                              21


<210> 438
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 2B spacer sequence

<400> 438
gactggactg ctgccacgtg c                                              21


<210> 439
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 3B spacer sequence

<400> 439
gctggactgc tgccacgtgc t                                              21


<210> 440
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 4B spacers sequence

<400> 440
gcacgtgctg ggctctgctg c                                              21


<210> 441
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 5B spacer sequence

<400> 441
gcagcagcag agcccagcac g                                              21


<210> 442
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> HEK site 4 spacer sequence

<400> 442
ggcactgcgg ctggaggtgg                                                20


<210> 443
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 2 spacer sequence

<400> 443
gaccccctcc accccgcctc                                                20


<210> 444
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> ZSCAN2 spacer sequence

<400> 444
gtgcggcaag agcttcagcc                                                20


<210> 445
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> ZSCAN2-21G spacer sequence

<400> 445
ggtgcggcaa gagcttcagc c                                              21


<210> 446
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EMX1 site 2 spacer sequence

<400> 446
gtcacctcca atgactaggg                                                20


<210> 447
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> EMX1 site 2-21G spacer sequence

<400> 447
ggtcacctcc aatgactagg g                                              21


<210> 448
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> DNMT1 site 4 spacer sequence

<400> 448
ggagtgaggg aaacggcccc                                                20


<210> 449
<211> 21
<212> DNA
<213> Artificial Sequence


<220> 
<223> DNMT1 site 4-21G spacer sequence

<400> 449
gggagtgagg gaaacggccc c                                              21


<210> 450
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 1 sequence with PAM

<400> 450
gcgatccagg tgctgcagaa ggg                                            23


<210> 451
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 2 sequence with PAM

<400> 451
gctgcagaag ggattccatg agg                                            23


<210> 452
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 3 sequence with PAM

<400> 452
tagggccttc gcgcacctca tgg                                            23


<210> 453
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 4 sequence with PAM

<400> 453
ccaaggtgaa agcggaagta ggg                                            23


<210> 454
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 5 sequence with PAM

<400> 454
tccaaggtga aagcggaagt agg                                            23


<210> 455
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 6 sequence with PAM

<400> 455
cgccgtctcc aaggtgaaag cgg                                            23


<210> 456
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 7 sequence with PAM

<400> 456
cctgctctct ctgcgcctgc tgg                                            23


<210> 457
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 8 sequence with PAM

<400> 457
ccagcaggcg cagagagagc agg                                            23


<210> 458
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 9 sequence with PAM

<400> 458
caggacgtca cagtgaccga ggg                                            23


<210> 459
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 10 sequence with PAM

<400> 459
ttagcgaact tccaggccct cgg                                            23


<210> 460
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 11 sequence with PAM

<400> 460
cgtcacagtg accgagggcc tgg                                            23


<210> 461
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> FANCF site 12 sequence with PAM

<400> 461
tccgggatta gcgaacttcc agg                                            23


<210> 462
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 1 sequence with PAM

<400> 462
cttcatgtac agagagccca ggg                                            23


<210> 463
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 3 sequence with PAM

<400> 463
aacagctaca tatttgggac tgg                                            23


<210> 464
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 4 sequence with PAM

<400> 464
gtcccaaata tgtagctgtt tgg                                            23


<210> 465
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 5 sequence with PAM

<400> 465
caaatatgta gctgtttggg agg                                            23


<210> 466
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 6 sequence with PAM

<400> 466
tttgggaggt cagaaatagg ggg                                            23


<210> 467
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 7 sequence with PAM

<400> 467
gggtgggggg agtttgctcc tgg                                            23


<210> 468
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 8 sequence with PAM

<400> 468
ctttgctagg aatattgaag ggg                                            23


<210> 469
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 9 sequence with PAM

<400> 469
ggcaggggaa ggcggagagc cgg                                            23


<210> 470
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 10 sequence with PAM

<400> 470
tctccccacc cgtccctgtc cgg                                            23


<210> 471
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 11 sequence with PAM

<400> 471
gagagccgga cagggacggg tgg                                            23


<210> 472
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 12 sequence with PAM

<400> 472
ggacagggac gggtggggag agg                                            23


<210> 473
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 1B sequence with PAM

<400> 473
ttctgcccag taggatgaac tgg                                            23


<210> 474
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 2B sequence with PAM

<400> 474
actggactgc tgccacgtgc tgg                                            23


<210> 475
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 3B sequence with PAM

<400> 475
ctggactgct gccacgtgct ggg                                            23


<210> 476
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 4B sequence with PAM

<400> 476
cacgtgctgg gctctgctgc tgg                                            23


<210> 477
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> mouseSprn site 5B sequence with PAM

<400> 477
cagcagcaga gcccagcacg tgg                                            23


<210> 478
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> HEK site 4 sequence with PAM

<400> 478
ggcactgcgg ctggaggtgg ggg                                            23


<210> 479
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> VEGFA site 2 sequence with PAM

<400> 479
gaccccctcc accccgcctc cgg                                            23


<210> 480
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> ZSCAN2 sequence with PAM

<400> 480
gtgcggcaag agcttcagcc ggg                                            23


<210> 481
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> ZSCAN2-21G sequence with PAM

<400> 481
gtgcggcaag agcttcagcc ggg                                            23


<210> 482
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EMX1 site 2 sequence with PAM

<400> 482
gtcacctcca atgactaggg tgg                                            23


<210> 483
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> EMX1 site 2-21G sequence with PAM

<400> 483
gtcacctcca atgactaggg tgg                                            23


<210> 484
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> DNMT1 site 4 sequence with PAM

<400> 484
ggagtgaggg aaacggcccc agg                                            23


<210> 485
<211> 23
<212> DNA
<213> Artificial Sequence


<220> 
<223> DNMT1 site 4-21G sequence with PAM

<400> 485
ggagtgaggg aaacggcccc agg                                            23


<210> 486
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 1 G19H mixed mismatched spacer sequence

<400> 486
ghgcacgggc agcttgccgg                                                20


<210> 487
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 1 C17D mixed mismatched spacer sequence

<400> 487
gggdacgggc agcttgccgg                                                20


<210> 488
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 1 C15D mixed mismatched spacer sequence

<400> 488
gggcadgggc agcttgccgg                                                20


<210> 489
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 2 A19B mixed mismatched spacer sequence

<400> 489
gbgctggacg gcgacgtaaa                                                20


<210> 490
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 2 C17D mixed mismatched spacer sequence

<400> 490
gagdtggacg gcgacgtaaa                                                20


<210> 491
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 2 G15H mixed mismatched spacer sequence

<400> 491
gagcthgacg gcgacgtaaa                                                20


<210> 492
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 3 T18V mixed mismatched spacer sequence

<400> 492
ggvggtgcag atgaacttca                                                20


<210> 493
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 3 G16H mixed mismatched spacer sequence

<400> 493
ggtghtgcag atgaacttca                                                20


<210> 494
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 3 G14H mixed mismatched spacer sequence

<400> 494
ggtggthcag atgaacttca                                                20


<210> 495
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 4 A18B mixed mismatched spacer sequence

<400> 495
ggbgcgcacc atcttcttca                                                20


<210> 496
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 4 C16D mixed mismatched spacer sequence

<400> 496
ggagdgcacc atcttcttca                                                20


<210> 497
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 4 C14D mixed mismatched spacer sequence

<400> 497
ggagcgdacc atcttcttca                                                20


<210> 498
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 5 T19V mixed mismatched spacer sequence

<400> 498
gvcgccctcg aacttcacct                                                20


<210> 499
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 5 G17H mixed mismatched spacer sequence

<400> 499
gtchccctcg aacttcacct                                                20


<210> 500
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 5 C15D mixed mismatched spacer sequence

<400> 500
gtcgcdctcg aacttcacct                                                20


<210> 501
<400> 501
000

<210> 502
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 6 C17D mixed mismatched spacer sequence

<400> 502
gcgdgatcac atggtcctgc                                                20


<210> 503
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 6 A15B mixed mismatched spacer sequence

<400> 503
gcgcgbtcac atggtcctgc                                                20


<210> 504
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 7 T19V mixed mismatched spacer sequence

<400> 504
gvccatgccg agagtgatcc                                                20


<210> 505
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 7 C17D mixed mismatched spacer sequence

<400> 505
gtcdatgccg agagtgatcc                                                20


<210> 506
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 7 T15V mixed mismatched spacer sequence

<400> 506
gtccavgccg agagtgatcc                                                20


<210> 507
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 8 C18D mixed mismatched spacer sequence

<400> 507
gcdgtcgtcc ttgaagaaga                                                20


<210> 508
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 8 T16V mixed mismatched spacer sequence

<400> 508
gccgvcgtcc ttgaagaaga                                                20


<210> 509
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 8 G14H mixed mismatched spacer sequence

<400> 509
gccgtchtcc ttgaagaaga                                                20


<210> 510
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 9 A18B mixed mismatched spacer sequence

<400> 510
gabgttcgag ggcgacaccc                                                20


<210> 511
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 9 T16V mixed mismatched spacer sequence

<400> 511
gaagvtcgag ggcgacaccc                                                20


<210> 512
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 9 C14D mixed mismatched spacer sequence

<400> 512
gaagttdgag ggcgacaccc                                                20


<210> 513
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 10 T19V mixed mismatched spacer sequence

<400> 513
gvctttgctc agggcggact                                                20


<210> 514
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 10 T17V mixed mismatched spacer sequence

<400> 514
gtcvttgctc agggcggact                                                20


<210> 515
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 10 T15V mixed mismatched spacer sequence

<400> 515
gtcttvgctc agggcggact                                                20


<210> 516
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 11 T18V mixed mismatched spacer sequence

<400> 516
ggvctttgct cagggcggac                                                20


<210> 517
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 11 T16V mixed mismatched spacer sequence

<400> 517
ggtcvttgct cagggcggac                                                20


<210> 518
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 11 T14V mixed mismatched spacer sequence

<400> 518
ggtcttvgct cagggcggac                                                20


<210> 519
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 12 G18H mixed mismatched spacer sequence

<400> 519
gthctcaggt agtggttgtc                                                20


<210> 520
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 12 T16V mixed mismatched spacer sequence

<400> 520
gtgcvcaggt agtggttgtc                                                20


<210> 521
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 12 A14B mixed mismatched spacer sequence

<400> 521
gtgctcbggt agtggttgtc                                                20


<210> 522
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 15 C18D mixed mismatched spacer sequence

<400> 522
ggdatcgccc tcgccctcgc                                                20


<210> 523
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 15 T16V mixed mismatched spacer sequence

<400> 523
ggcavcgccc tcgccctcgc                                                20


<210> 524
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 15 G14H mixed mismatched spacer sequence

<400> 524
ggcatchccc tcgccctcgc                                                20


<210> 525
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 16 C18D mixed mismatched spacer sequence

<400> 525
ggdgagggcg atgccaccta                                                20


<210> 526
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 16 A16B mixed mismatched spacer sequence

<400> 526
ggcgbgggcg atgccaccta                                                20


<210> 527
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 16 G14H mixed mismatched spacer sequence

<400> 527
ggcgaghgcg atgccaccta                                                20


<210> 528
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 24 T19V mixed mismatched spacer sequence

<400> 528
gvcgtgctgc ttcatgtggt                                                20


<210> 529
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 24 G17H mixed mismatched spacer sequence

<400> 529
gtchtgctgc ttcatgtggt                                                20


<210> 530
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 24 G15H mixed mismatched spacer sequence

<400> 530
gtcgthctgc ttcatgtggt                                                20


<210> 531
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 43 A18B mixed mismatched spacer sequence

<400> 531
gabgggcatc gacttcaagg                                                20


<210> 532
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 43 G16H mixed mismatched spacer sequence

<400> 532
gagghgcatc gacttcaagg                                                20


<210> 533
<211> 20
<212> DNA
<213> Artificial Sequence


<220> 
<223> EGFP site 43 C14D mixed mismatched spacer sequence

<400> 533
gaggggdatc gacttcaagg                                                20


<210> 534
<211> 8077
<212> DNA
<213> Artificial Sequence


<220> 
<223> pX330-Flag-WT SpCas9 (without sgRNA; with silent mutations)

<220> 
<221> misc_feature
<222> 826..891
<223> /note="3xFLAG tag "

<220> 
<221> misc_feature
<222> 898..928
<223> /note="NLS"

<220> 
<221> misc_feature
<222> 943..5043
<223> /note="Human codon optimized S. pyogenes Cas9 "

<400> 534
ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac       60

ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg      120

gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg      180

ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc      240

ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc      300

gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt      360

ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg      420

cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg      480

gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg      540

cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgac gctgccttcg      600

ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt      660

actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa      720

gaggtaaggg tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca      780

cctgcctgaa atcacttttt ttcaggttgg accggtgcca ccatggacta taaggaccac      840

gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca      900

aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc      960

ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg     1020

cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg     1080

atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc     1140

gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc     1200

aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg     1260

gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc     1320

taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac     1380

aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac     1440

ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag     1500

ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac     1560

gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc     1620

cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc     1680

ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc     1740

aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc     1800

gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga     1860

gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag     1920

caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac     1980

aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc     2040

agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag     2100

gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac     2160

ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa     2220

gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc     2280

atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga     2340

aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc     2400

gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg     2460

ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg     2520

aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc     2580

atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac     2640

tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc     2700

aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg     2760

gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag     2820

gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg     2880

atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc     2940

aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc     3000

ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac     3060

atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg     3120

gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc     3180

gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac     3240

cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc     3300

atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag     3360

aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa     3420

ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg     3480

aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc     3540

gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg     3600

aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc     3660

ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc     3720

acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac     3780

aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg     3840

aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcg     3900

tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag     3960

ttcgtgtacg gcgactacaa ggtgtacgac gtacggaaga tgatcgccaa gagcgagcag     4020

gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag     4080

accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc     4140

gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg     4200

agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa     4260

gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac     4320

cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc     4380

aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc     4440

atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac     4500

aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa     4560

aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc     4620

ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc     4680

tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac     4740

gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg     4800

gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag     4860

aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt     4920

gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg     4980

atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc     5040

gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gtaagaattc     5100

ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc     5160

cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa     5220

atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg     5280

ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc     5340

aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg     5400

ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc     5460

gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg     5520

gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag     5580

cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc     5640

cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc     5700

tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa     5760

aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg     5820

ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac     5880

actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta     5940

ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac     6000

gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca     6060

gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc     6120

cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc     6180

atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt     6240

catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac     6300

ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc     6360

ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt     6420

cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct     6480

ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga     6540

tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag     6600

cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca     6660

actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga     6720

aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag     6780

tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc     6840

ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa     6900

tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt     6960

gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg     7020

gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt     7080

tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg     7140

gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat     7200

ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact     7260

gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa     7320

aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt     7380

ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt     7440

ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg     7500

tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca     7560

gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt     7620

agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga     7680

taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc     7740

gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact     7800

gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga     7860

caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg     7920

aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt     7980

tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt     8040

acggttcctg gccttttgct ggccttttgc tcacatg                              8077


<210> 535
<211> 64
<212> DNA
<213> Artificial Sequence


<220> 
<223> 9214-SpCas9_bact_exp-for forward primer

<400> 535
aaaaaaacta gtgaaaacct gtatttccag ggagcagcct cgatggacta taaggaccac     60

gacg                                                                  64


<210> 536
<211> 33
<212> DNA
<213> Artificial Sequence


<220> 
<223> 9214-SpCas9_bact_exp-rev reverse primer

<400> 536
aaaaaagcgg ccgcaacaga tggctggcaa cta                                  33


<210> 537
<211> 94
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8717for forward primer

<400> 537
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag     60

cgggttcgtg tacggcgacg gcaaggtgta cgat                                 94


<210> 538
<211> 94
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8717rev reverse primer

<400> 538
gtacatcgta caccttgccg tcgccgtaca cgaacccgct ttccagctta gggtactttt     60

tgatcagggc ggttcccacg acggcgttca ggta                                 94


<210> 539
<211> 67
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8727for forward primer

<400> 539
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agggtaaggt     60

gtacgat                                                               67


<210> 540
<211> 67
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8727rev reverse primer

<400> 540
gtacatcgta caccttaccc ttagggtact ttttgatcag ggcggttccc acgacggcgt     60

tcaggta                                                               67


<210> 541
<211> 55
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8724for forward primer

<400> 541
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta aggat          55


<210> 542
<211> 55
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8724rev reverse primer

<400> 542
gtacatcctt agggtacttt ttgatcaggg cggttcccac gacggcgttc aggta          55


<210> 543
<211> 58
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8725for forward primer

<400> 543
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agggtgat       58


<210> 544
<211> 58
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8725rev reverse primer

<400> 544
gtacatcacc cttagggtac tttttgatca gggcggttcc cacgacggcg ttcaggta       58


<210> 545
<211> 61
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8726for forward primer

<400> 545
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agggtaaaga     60

t                                                                     61


<210> 546
<211> 61
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8726rev reverse primer

<400> 546
gtacatcttt acccttaggg tactttttga tcagggcggt tcccacgacg gcgttcaggt     60

a                                                                     61


<210> 547
<211> 67
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8718for forward primer

<400> 547
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgaaggt     60

gtacgat                                                               67


<210> 548
<211> 67
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8718rev reverse primer

<400> 548
gtacatcgta caccttcagc ttagggtact ttttgatcag ggcggttccc acgacggcgt     60

tcaggta                                                               67


<210> 549
<211> 70
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8721for forward primer

<400> 549
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtaa     60

ggtgtacgat                                                            70


<210> 550
<211> 70
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8721rev reverse primer

<400> 550
gtacatcgta caccttaccc agcttagggt actttttgat cagggcggtt cccacgacgg     60

cgttcaggta                                                            70


<210> 551
<211> 70
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8730for forward primer

<400> 551
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgcctaa     60

ggtgtacgat                                                            70


<210> 552
<211> 70
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8730rev reverse primer

<400> 552
gtacatcgta caccttaggc agcttagggt actttttgat cagggcggtt cccacgacgg     60

cgttcaggta                                                            70


<210> 553
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8719for forward primer

<400> 553
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtgg     60

caaggtgtac gat                                                        73


<210> 554
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8719rev reverse primer

<400> 554
gtacatcgta caccttgcca cccagcttag ggtacttttt gatcagggcg gttcccacga     60

cggcgttcag gta                                                        73


<210> 555
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8729for forward primer

<400> 555
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtcc     60

taaggtgtac gat                                                        73


<210> 556
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8729rev reverse primer

<400> 556
gtacatcgta caccttagga cccagcttag ggtacttttt gatcagggcg gttcccacga     60

cggcgttcag gta                                                        73


<210> 557
<211> 76
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8741for forward primer

<400> 557
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtgg     60

cggaaaggtg tacgat                                                     76


<210> 558
<211> 76
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8741rev reverse primer

<400> 558
gtacatcgta cacctttccg ccacccagct tagggtactt tttgatcagg gcggttccca     60

cgacggcgtt caggta                                                     76


<210> 559
<211> 79
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8742for forward primer

<400> 559
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtgg     60

cggaggtaag gtgtacgat                                                  79


<210> 560
<211> 79
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8742rev reverse primer

<400> 560
gtacatcgta caccttacct ccgccaccca gcttagggta ctttttgatc agggcggttc     60

ccacgacggc gttcaggta                                                  79


<210> 561
<211> 70
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8738for forward primer

<400> 561
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtgg     60

cgtgtacgat                                                            70


<210> 562
<211> 70
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8738rev reverse primer

<400> 562
gtacatcgta cacgccaccc agcttagggt actttttgat cagggcggtt cccacgacgg     60

cgttcaggta                                                            70


<210> 563
<211> 67
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8739for forward primer

<400> 563
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtgg     60

ctacgat                                                               67


<210> 564
<211> 67
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8739rev reverse primer

<400> 564
gtacatcgta gccacccagc ttagggtact ttttgatcag ggcggttccc acgacggcgt     60

tcaggta                                                               67


<210> 565
<211> 64
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8740for forward primer

<400> 565
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgggtgg     60

cgat                                                                  64


<210> 566
<211> 64
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8740rev reverse primer

<400> 566
gtacatcgcc acccagctta gggtactttt tgatcagggc ggttcccacg acggcgttca     60

ggta                                                                  64


<210> 567
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8756for forward primer

<400> 567
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgtctag     60

caaggtgtac gat                                                        73


<210> 568
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8756rev reverse primer

<400> 568
gtacatcgta caccttgcta gacagcttag ggtacttttt gatcagggcg gttcccacga     60

cggcgttcag gta                                                        73


<210> 569
<211> 64
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8728for forward primer

<400> 569
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctgtctag     60

cgat                                                                  64


<210> 570
<211> 64
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8728rev reverse primer

<400> 570
gtacatcgct agacagctta gggtactttt tgatcagggc ggttcccacg acggcgttca     60

ggta                                                                  64


<210> 571
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8722for forward primer

<400> 571
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag     60

caaggtgtac gat                                                        73


<210> 572
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8722rev reverse primmer

<400> 572
gtacatcgta caccttgctt tccagcttag ggtacttttt gatcagggcg gttcccacga     60

cggcgttcag gta                                                        73


<210> 573
<211> 76
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8723for forward primer

<400> 573
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag     60

cggtaaggtg tacgat                                                     76


<210> 574
<211> 76
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8723rev reverse primer

<400> 574
gtacatcgta caccttaccg ctttccagct tagggtactt tttgatcagg gcggttccca     60

cgacggcgtt caggta                                                     76


<210> 575
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8752for forward primer

<400> 575
cgcgtacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtaccctg cgctgggtgg     60

caaggtgtac gat                                                        73


<210> 576
<211> 73
<212> DNA
<213> Artificial Sequence


<220> 
<223> 8752rev reverse primer

<400> 576
gtacatcgta caccttgcca cccagcgcag ggtacttttt gatcagggcg gttcccacga     60

cggcgttcag gta                                                        73


<210> 577
<211> 24
<212> DNA
<213> Artificial Sequence


<220> 
<223> HF+primer_rev reverse primer

<400> 577
gaagccgccg tacttcttag gatc                                           24


<210> 578
<211> 27
<212> DNA
<213> Artificial Sequence


<220> 
<223> HF+primer_for forward primer

<400> 578
gaggaaaacg aggacattct ggaagat                                        27


<210> 579
<211> 33
<212> DNA
<213> Artificial Sequence


<220> 
<223> A661Rfor forward primer

<400> 579
cggctggggc aggctgagcc ggaagctgat caa                                  33


<210> 580
<211> 28
<212> DNA
<213> Artificial Sequence


<220> 
<223> A661Rrev reverse primer

<400> 580
cggctcagcc tgccccagcc ggtgtatc                                       28


<210> 581
<211> 35
<212> DNA
<213> Artificial Sequence


<220> 
<223> A695Qfor forward primer

<400> 581
aaacttcatg cagctgatcc acgacgacag cctga                                35


<210> 582
<211> 39
<212> DNA
<213> Artificial Sequence


<220> 
<223> A695Qrev reverse primer

<400> 582
cgtggatcag ctgcatgaag tttctgttgg cgaagccgt                            39


<210> 583
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> A926Qrev reverse primer

<400> 583
gctttgtgat ctgccgggtt tccaccagct gt                                   32


<210> 584
<211> 33
<212> DNA
<213> Artificial Sequence


<220> 
<223> A926Qfor forward primer

<400> 584
ggaaacccgg cagatcacaa agcacgtggc aca                                  33


