                         SEQUENCE LISTING

<110>  REGENTS OF THE UNIVERSITY OF CALIFORNIA 
       Julian HALMAI
       Kyle FINK
       Jennifer WALDO
 
<120>  USE OF A SPLIT dCAS FUSION PROTEIN SYSTEM FOR EPIGENETIC EDITING

<130>  081906-1295346-242910PC

<140>  PCT/US2022/025412
<141>  2022-04-19

<150>  63/177,523
<151>  2021-04-21

<160>  52    

<170>  PatentIn version 3.5

<210>  1
<211>  1367
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Description of sequence: SpdCas9 amino acid sequence

<400>  1

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  2
<211>  154
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Description of sequence: RmaDnaB amino acid sequence

<400>  2

Cys Leu Ala Gly Asp Thr Leu Ile Thr Leu Ala Asp Gly Arg Arg Val 
1               5                   10                  15      


Pro Ile Arg Glu Leu Val Ser Gln Gln Asn Phe Ser Val Trp Ala Leu 
            20                  25                  30          


Asn Pro Gln Thr Tyr Arg Leu Glu Arg Ala Arg Val Ser Arg Ala Phe 
        35                  40                  45              


Cys Thr Gly Ile Lys Pro Val Tyr Arg Leu Thr Thr Arg Leu Gly Arg 
    50                  55                  60                  


Ser Ile Arg Ala Thr Ala Asn His Arg Phe Leu Thr Pro Gln Gly Trp 
65                  70                  75                  80  


Lys Arg Val Asp Glu Leu Gln Pro Gly Asp Tyr Leu Ala Leu Pro Arg 
                85                  90                  95      


Arg Ile Pro Thr Ala Ser Met Ala Ala Ala Cys Pro Glu Leu Arg Gln 
            100                 105                 110         


Leu Ala Gln Ser Asp Val Tyr Trp Asp Pro Ile Val Ser Ile Glu Pro 
        115                 120                 125             


Asp Gly Val Glu Glu Val Phe Asp Leu Thr Val Pro Gly Pro His Asn 
    130                 135                 140                 


Phe Val Ala Asn Asp Ile Ile Ala His Asn 
145                 150                 


<210>  3
<211>  814
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Amino acid sequence for SpdCas9N-RmaDnaB

<400>  3

Asp Lys Lys Tyr Ser Ile Gly Leu Ala 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 Cys Leu Ala Gly Asp Thr Leu Ile 
705                 710                 715                 720 


Thr Leu Ala Asp Gly Arg Arg Val Pro Ile Arg Glu Leu Val Ser Gln 
                725                 730                 735     


Gln Asn Phe Ser Val Trp Ala Leu Asn Pro Gln Thr Tyr Arg Leu Glu 
            740                 745                 750         


Arg Ala Arg Val Ser Arg Ala Phe Cys Thr Gly Ile Lys Pro Val Tyr 
        755                 760                 765             


Arg Leu Thr Thr Arg Leu Gly Arg Ser Ile Arg Ala Thr Ala Asn His 
    770                 775                 780                 


Arg Phe Leu Thr Pro Gln Gly Trp Lys Arg Val Asp Glu Leu Gln Pro 
785                 790                 795                 800 


Gly Asp Tyr Leu Ala Leu Pro Arg Arg Ile Pro Thr Ala Ser 
                805                 810                 


<210>  4
<211>  711
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Amino acid sequence for RmaDnaB-SpdCas9C

<400>  4

Met Ala Ala Ala Cys Pro Glu Leu Arg Gln Leu Ala Gln Ser Asp Val 
1               5                   10                  15      


Tyr Trp Asp Pro Ile Val Ser Ile Glu Pro Asp Gly Val Glu Glu Val 
            20                  25                  30          


Phe Asp Leu Thr Val Pro Gly Pro His Asn Phe Val Ala Asn Asp Ile 
        35                  40                  45              


Ile Ala His Asn Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala 
    50                  55                  60                  


Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val 
65                  70                  75                  80  


Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu 
                85                  90                  95      


Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly 
            100                 105                 110         


Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys 
        115                 120                 125             


Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln 
    130                 135                 140                 


Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp 
145                 150                 155                 160 


Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp 
                165                 170                 175     


Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp 
            180                 185                 190         


Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn 
        195                 200                 205             


Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln 
    210                 215                 220                 


Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr 
225                 230                 235                 240 


Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile 
                245                 250                 255     


Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln 
            260                 265                 270         


Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu 
        275                 280                 285             


Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp 
    290                 295                 300                 


Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr 
305                 310                 315                 320 


His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu 
                325                 330                 335     


Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr 
            340                 345                 350         


Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile 
        355                 360                 365             


Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe 
    370                 375                 380                 


Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro 
385                 390                 395                 400 


Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly 
                405                 410                 415     


Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn 
            420                 425                 430         


Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser 
        435                 440                 445             


Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp 
    450                 455                 460                 


Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr 
465                 470                 475                 480 


Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu 
                485                 490                 495     


Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser 
            500                 505                 510         


Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu 
        515                 520                 525             


Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu 
    530                 535                 540                 


Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln 
545                 550                 555                 560 


Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu Tyr 
                565                 570                 575     


Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu 
            580                 585                 590         


Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr Leu Asp Glu Ile 
        595                 600                 605             


Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala 
    610                 615                 620                 


Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro 
625                 630                 635                 640 


Ile Arg Glu Gln Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn 
                645                 650                 655     


Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg 
            660                 665                 670         


Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His 
        675                 680                 685             


Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu 
    690                 695                 700                 


Gly Gly Asp Ser Arg Ala Asp 
705                 710     


<210>  5
<211>  1525
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Description of sequence: dCas9-intein-all in one

<400>  5

Asp Lys Lys Tyr Ser Ile Gly Leu Ala 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 Cys Leu Ala Gly Asp Thr Leu Ile 
705                 710                 715                 720 


Thr Leu Ala Asp Gly Arg Arg Val Pro Ile Arg Glu Leu Val Ser Gln 
                725                 730                 735     


Gln Asn Phe Ser Val Trp Ala Leu Asn Pro Gln Thr Tyr Arg Leu Glu 
            740                 745                 750         


Arg Ala Arg Val Ser Arg Ala Phe Cys Thr Gly Ile Lys Pro Val Tyr 
        755                 760                 765             


Arg Leu Thr Thr Arg Leu Gly Arg Ser Ile Arg Ala Thr Ala Asn His 
    770                 775                 780                 


Arg Phe Leu Thr Pro Gln Gly Trp Lys Arg Val Asp Glu Leu Gln Pro 
785                 790                 795                 800 


Gly Asp Tyr Leu Ala Leu Pro Arg Arg Ile Pro Thr Ala Ser Met Ala 
                805                 810                 815     


Ala Ala Cys Pro Glu Leu Arg Gln Leu Ala Gln Ser Asp Val Tyr Trp 
            820                 825                 830         


Asp Pro Ile Val Ser Ile Glu Pro Asp Gly Val Glu Glu Val Phe Asp 
        835                 840                 845             


Leu Thr Val Pro Gly Pro His Asn Phe Val Ala Asn Asp Ile Ile Ala 
    850                 855                 860                 


His Asn Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu 
865                 870                 875                 880 


Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val 
                885                 890                 895     


Val Asp Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile 
            900                 905                 910         


Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys 
        915                 920                 925             


Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu 
    930                 935                 940                 


Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln 
945                 950                 955                 960 


Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr 
                965                 970                 975     


Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp 
            980                 985                 990         


Ala Ile Val Pro Gln Ser Phe Leu  Lys Asp Asp Ser Ile  Asp Asn Lys 
        995                 1000                 1005             


Val Leu  Thr Arg Ser Asp Lys  Asn Arg Gly Lys Ser  Asp Asn Val 
    1010                 1015                 1020             


Pro Ser  Glu Glu Val Val Lys  Lys Met Lys Asn Tyr  Trp Arg Gln 
    1025                 1030                 1035             


Leu Leu  Asn Ala Lys Leu Ile  Thr Gln Arg Lys Phe  Asp Asn Leu 
    1040                 1045                 1050             


Thr Lys  Ala Glu Arg Gly Gly  Leu Ser Glu Leu Asp  Lys Ala Gly 
    1055                 1060                 1065             


Phe Ile  Lys Arg Gln Leu Val  Glu Thr Arg Gln Ile  Thr Lys His 
    1070                 1075                 1080             


Val Ala  Gln Ile Leu Asp Ser  Arg Met Asn Thr Lys  Tyr Asp Glu 
    1085                 1090                 1095             


Asn Asp  Lys Leu Ile Arg Glu  Val Lys Val Ile Thr  Leu Lys Ser 
    1100                 1105                 1110             


Lys Leu  Val Ser Asp Phe Arg  Lys Asp Phe Gln Phe  Tyr Lys Val 
    1115                 1120                 1125             


Arg Glu  Ile Asn Asn Tyr His  His Ala His Asp Ala  Tyr Leu Asn 
    1130                 1135                 1140             


Ala Val  Val Gly Thr Ala Leu  Ile Lys Lys Tyr Pro  Lys Leu Glu 
    1145                 1150                 1155             


Ser Glu  Phe Val Tyr Gly Asp  Tyr Lys Val Tyr Asp  Val Arg Lys 
    1160                 1165                 1170             


Met Ile  Ala Lys Ser Glu Gln  Glu Ile Gly Lys Ala  Thr Ala Lys 
    1175                 1180                 1185             


Tyr Phe  Phe Tyr Ser Asn Ile  Met Asn Phe Phe Lys  Thr Glu Ile 
    1190                 1195                 1200             


Thr Leu  Ala Asn Gly Glu Ile  Arg Lys Arg Pro Leu  Ile Glu Thr 
    1205                 1210                 1215             


Asn Gly  Glu Thr Gly Glu Ile  Val Trp Asp Lys Gly  Arg Asp Phe 
    1220                 1225                 1230             


Ala Thr  Val Arg Lys Val Leu  Ser Met Pro Gln Val  Asn Ile Val 
    1235                 1240                 1245             


Lys Lys  Thr Glu Val Gln Thr  Gly Gly Phe Ser Lys  Glu Ser Ile 
    1250                 1255                 1260             


Leu Pro  Lys Arg Asn Ser Asp  Lys Leu Ile Ala Arg  Lys Lys Asp 
    1265                 1270                 1275             


Trp Asp  Pro Lys Lys Tyr Gly  Gly Phe Asp Ser Pro  Thr Val Ala 
    1280                 1285                 1290             


Tyr Ser  Val Leu Val Val Ala  Lys Val Glu Lys Gly  Lys Ser Lys 
    1295                 1300                 1305             


Lys Leu  Lys Ser Val Lys Glu  Leu Leu Gly Ile Thr  Ile Met Glu 
    1310                 1315                 1320             


Arg Ser  Ser Phe Glu Lys Asn  Pro Ile Asp Phe Leu  Glu Ala Lys 
    1325                 1330                 1335             


Gly Tyr  Lys Glu Val Lys Lys  Asp Leu Ile Ile Lys  Leu Pro Lys 
    1340                 1345                 1350             


Tyr Ser  Leu Phe Glu Leu Glu  Asn Gly Arg Lys Arg  Met Leu Ala 
    1355                 1360                 1365             


Ser Ala  Gly Glu Leu Gln Lys  Gly Asn Glu Leu Ala  Leu Pro Ser 
    1370                 1375                 1380             


Lys Tyr  Val Asn Phe Leu Tyr  Leu Ala Ser His Tyr  Glu Lys Leu 
    1385                 1390                 1395             


Lys Gly  Ser Pro Glu Asp Asn  Glu Gln Lys Gln Leu  Phe Val Glu 
    1400                 1405                 1410             


Gln His  Lys His Tyr Leu Asp  Glu Ile Ile Glu Gln  Ile Ser Glu 
    1415                 1420                 1425             


Phe Ser  Lys Arg Val Ile Leu  Ala Asp Ala Asn Leu  Asp Lys Val 
    1430                 1435                 1440             


Leu Ser  Ala Tyr Asn Lys His  Arg Asp Lys Pro Ile  Arg Glu Gln 
    1445                 1450                 1455             


Ala Glu  Asn Ile Ile His Leu  Phe Thr Leu Thr Asn  Leu Gly Ala 
    1460                 1465                 1470             


Pro Ala  Ala Phe Lys Tyr Phe  Asp Thr Thr Ile Asp  Arg Lys Arg 
    1475                 1480                 1485             


Tyr Thr  Ser Thr Lys Glu Val  Leu Asp Ala Thr Leu  Ile His Gln 
    1490                 1495                 1500             


Ser Ile  Thr Gly Leu Tyr Glu  Thr Arg Ile Asp Leu  Ser Gln Leu 
    1505                 1510                 1515             


Gly Gly  Asp Ser Arg Ala Asp  
    1520                 1525 


<210>  6
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: IDT human codon optimized SpdCas9 sequence

<400>  6
gacaagaagt actccattgg gctcgctatc ggcacaaaca gcgtcggctg ggccgtcatt       60

acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac      120

agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc      180

acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac      240

ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg      300

gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat      360

atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag      420

cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg      480

atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc      540

gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc      600

aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg      660

ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt      720

atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat      780

gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag      840

atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg      900

ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg      960

atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag     1020

caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga     1080

tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa     1140

aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa     1200

cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct     1260

atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag     1320

aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga     1380

ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc     1440

gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat     1500

ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat     1560

aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct     1620

ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg     1680

aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc     1740

ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt     1800

aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc     1860

acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat     1920

ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg     1980

ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat     2040

tttcttaagt ccgatggatt tgccaacagg aacttcatgc agttgatcca tgatgactct     2100

ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac     2160

gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt     2220

aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc     2280

gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg     2340

aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt     2400

gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac     2460

atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatgctatc     2520

gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat     2580

aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat     2640

tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact     2700

aaggctgaac gaggtggcct gtctgagttg gataaagccg gcttcatcaa aaggcagctt     2760

gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc     2820

aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag     2880

ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac     2940

caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat     3000

cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg     3060

atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat     3120

attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca     3180

cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg     3240

acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag     3300

accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca     3360

cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac     3420

agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag     3480

gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt     3540

ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac     3600

tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag     3660

aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac     3720

tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa     3780

cacaaacact accttgatga gatcatcgag caaataagcg agttctccaa aagagtgatc     3840

ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc     3900

atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct     3960

gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag     4020

gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac     4080

ctctctcagc tcggtggaga c                                               4101


<210>  7
<211>  306
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: N-intein IDT codon optimized DNA sequence (306bp)

<400>  7
tgtcttgcgg gggatactct cataaccctt gccgacgggc ggcgcgtacc tattagggag       60

ttggtgtcac agcagaactt ctccgtttgg gcacttaacc cacaaacgta tcgcttggag      120

cgggcgcggg ttagtagggc cttctgtaca ggtataaaac ccgtgtatag gctcaccact      180

aggcttggcc ggtccatccg agccacggcg aaccatcgct ttttgactcc acaaggatgg      240

aagagggttg acgaattgca gcctggagat tacctggctc tgccccggcg catacctacg      300

gctagt                                                                 306


<210>  8
<211>  156
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: C-intein IDT codon optimized sequence (156bp)

<400>  8
atggcggcgg cctgcccaga gcttcgccag ctggcgcaaa gcgacgtata ttgggaccca       60

atagtaagca tagaaccaga cggtgtcgaa gaggtgtttg acctgacggt gccaggccct      120

cacaacttcg ttgctaacga catcatagcc cataat                                156


<210>  9
<211>  701
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: CMV promoter sequence

<400>  9
atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag       60

ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct      120

gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc      180

caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg      240

cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat      300

ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca      360

tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc      420

gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga      480

gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat      540

tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag      600

tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc      660

gggaccgatc cagcctccgg actctagagg atcgaaccct t                          701


<210>  10
<211>  249
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: U6 promoter sequence

<400>  10
gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag       60

ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga      120

aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat      180

atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga      240

cgaaacacc                                                              249


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

<220>
<223>  Description of sequence: Tetrameric repeat sequence

<400>  11

Asp Ala Leu Asp Asp Phe Asp Leu Asp Met Leu 
1               5                   10      


<210>  12
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  12
agagcatcgg accgaagcgg                                                   20


<210>  13
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  13
gggggagaac atactcgggg                                                   20


<210>  14
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  14
cccaggttgc tagggcttgg                                                   20


<210>  15
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  15
atcgcctgaa acttgtccgg                                                   20


<210>  16
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  16
cgaaagggtg tgaaagaggg                                                   20


<210>  17
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  17
tggggaaggt aaagcggcga                                                   20


<210>  18
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  18
agagcatcgg accgaagcgg                                                   20


<210>  19
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  19
gggggagaac atactcgggg                                                   20


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

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  20
cccaggttgc tagggcttgg                                                   20


<210>  21
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  21
atcgcctgaa acttgtccgg                                                   20


<210>  22
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  22
cgaaagggtg tgaaagaggg                                                   20


<210>  23
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  23
tggggaaggt aaagcggcga                                                   20


<210>  24
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  24
gcttgagcaa tttcggaccc                                                   20


<210>  25
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  25
tgtgtctctt gctggtaccg                                                   20


<210>  26
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  26
tgagcctgtg ccagaggata                                                   20


<210>  27
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  27
tcaactttga ttgccaagtg ca                                                22


<210>  28
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  28
gagcagttct ggaaccaacc                                                   20


<210>  29
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  29
ttgaggccga agagagatgt                                                   20


<210>  30
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  30
ccttgtggaa tttgggtcat                                                   20


<210>  31
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  31
tcaaatgcag gcacttagaa t                                                 21


<210>  32
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  32
caaggaaaaa gagaagcaag ga                                                22


<210>  33
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  33
attttaatgg ctggctttgg                                                   20


<210>  34
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  34
ttttagttta ggttgttagg gtttg                                             25


<210>  35
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  35
taaaaaaaca cctcaaattt taccc                                             25


<210>  36
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  36
tcatcctcct tggaaacccg                                                   20


<210>  37
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  37
gtcatcgccc aaccagtaca                                                   20


<210>  38
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  38
agcagcagca atggacttcg                                                   20


<210>  39
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  39
agaaatacag gatggaggat ggt                                               23


<210>  40
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  40
aagcgcttcc tcctcattgg                                                   20


<210>  41
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  41
aaagcacctc aggttttgcc                                                   20


<210>  42
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  42
agctgttgat tggctgcttt                                                   20


<210>  43
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  43
ttcaaattcc gcccactaaa                                                   20


<210>  44
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  44
cacctcccag cttcactctc                                                   20


<210>  45
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  45
ctgcgggttc atctagttcc                                                   20


<210>  46
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  46
acaaccagca ttcgatccat                                                   20


<210>  47
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  47
gctgtcggaa ttgggtactg ttt                                               23


<210>  48
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  48
gctgtcggaa ttgggtactg ttg                                               23


<210>  49
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  49
aactcttact tggcgctccc                                                   20


<210>  50
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  50
ctgtccatcg ctaagctccc                                                   20


<210>  51
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  51
aatcccatca ccatcttcca                                                   20


<210>  52
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Description of sequence: Nucleotide target sequence

<400>  52
ctccatggtg gtgaagacg                                                    19


