                         SEQUENCE LISTING

<110>  Tsinghua University
 
<120>  Cas9 variants and application thereof

<130>  PIDC3181386P

<160>  137   

<170>  PatentIn version 3.3

<210>  1
<211>  981
<212>  PRT
<213>  Artificial

<220>
<223>   first backbone region of SaCas9-KKH variant

<400>  1

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val 
            980     


<210>  2
<211>  59
<212>  PRT
<213>  Artificial

<220>
<223>  second backbone region of SaCas9-KKH variant

<400>  2

Asn Met Ile Asp Ile Thr Tyr Arg Glu Tyr Leu Glu Asn Met Asn Asp 
1               5                   10                  15      


Lys Arg Pro Pro His Ile Ile Lys Thr Ile Ala Ser Lys Thr Gln Ser 
            20                  25                  30          


Ile Lys Lys Tyr Ser Thr Asp Ile Leu Gly Asn Leu Tyr Glu Val Lys 
        35                  40                  45              


Ser Lys Lys His Pro Gln Ile Ile Lys Lys Gly 
    50                  55                  


<210>  3
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 10 (v10)

<400>  3

Arg Ser Asp Ser Ser Pro Arg Glu Asn Arg Leu Glu Val 
1               5                   10              


<210>  4
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 12 (v12)

<400>  4

Lys Gly Asp Ala Met Pro Arg Gly Asn Lys Ile Glu Ile 
1               5                   10              


<210>  5
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 13 (v13)

<400>  5

Thr Ala Thr Asn Asn Asp Lys Ser Asn Lys Ile Glu Val 
1               5                   10              


<210>  6
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 14 (v14)

<400>  6

Leu Gly Asp Ala Asn Ser Arg Gln Asn Ile Leu Glu Ala 
1               5                   10              


<210>  7
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 15 (v15)

<400>  7

Ile Gly Val Asn His Asp Glu Gly Asn Arg Ile Glu Met 
1               5                   10              


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

<220>
<223>  PAM interaction region of Cas 9 variant 16 (v16)

<400>  8

Ile Gly Val Asn Ser Asp Lys Asn Asn Leu Ile Glu Val 
1               5                   10              


<210>  9
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 17 (v17)

<400>  9

Ile Gly Val Asn Asn Ser Thr Arg Asn Ile Val Glu Leu 
1               5                   10              


<210>  10
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 18 (v18)

<400>  10

Arg Gly Asp Asn Asn Pro Arg Gln Asn Lys Leu Glu Val 
1               5                   10              


<210>  11
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 19 (v19)

<400>  11

Ile Gly Val Asn Asn Asp Lys Asn Asn Val Ile Glu Leu 
1               5                   10              


<210>  12
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 20 (v20)

<400>  12

Ile Gly Ile Asn Asp Asn Lys His Asn Lys Ile Glu Leu 
1               5                   10              


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

<220>
<223>  PAM interaction region of Cas 9 variant 21 (v21)

<400>  13

Ile Gly Val Asn Ser Asp Asp Arg Asn Ile Ile Glu Leu 
1               5                   10              


<210>  14
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 22 (v22)

<400>  14

Ile Gly Val Asn Asp Ser Glu Lys Asn Lys Ile Gln Leu 
1               5                   10              


<210>  15
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 23 (v23)

<400>  15

Lys Cys Ile Asn Asn Glu Lys Thr His Arg Ile Glu Ile 
1               5                   10              


<210>  16
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 24 (v24)

<400>  16

Ile Gly Val Asn His Asp Lys Thr Asn Arg Ile Glu Cys 
1               5                   10              


<210>  17
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 25 (v25)

<400>  17

Ile Gly Val Asn Glu Asp Lys Arg Asn Ile Ile Glu Leu 
1               5                   10              


<210>  18
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 26 (v26)

<400>  18

Arg Gly Asp Ser Ser Pro Arg Glu Asn Arg Phe Glu Val 
1               5                   10              


<210>  19
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 27 (v27)

<400>  19

Arg Gly Asp Asn Asp Pro Lys Ala Asn Lys Ile Glu Val 
1               5                   10              


<210>  20
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 28 (v28)

<400>  20

Ile Gly Val Asn Ala Glu Lys Arg Asn Thr Ile Glu Val 
1               5                   10              


<210>  21
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 29 (v29)

<400>  21

Ile Gly Val Asn Asp Asp Ala Lys Asn Thr Leu Glu Leu 
1               5                   10              


<210>  22
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 30 (v30)

<400>  22

Val Gly Val Asn Asn Asp Ser Val Asn Arg Val Glu Leu 
1               5                   10              


<210>  23
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 31 (v31)

<400>  23

Val Gly Val Asn Asn Asp Thr Arg Asn Val Val Glu Leu 
1               5                   10              


<210>  24
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 32 (v32)

<400>  24

Val Gly Val Asn Asn Asp Ser Arg Asn Val Val Glu Leu 
1               5                   10              


<210>  25
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 33 (v33)

<400>  25

Arg Gly Asp Ser Met Pro Arg Gln Asn Lys Ile Glu Met 
1               5                   10              


<210>  26
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 34 (v34)

<400>  26

Arg Gly Asp Ala Met Pro Arg Asp Asn Lys Ile Glu Val 
1               5                   10              


<210>  27
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 35 (v35)

<400>  27

Ile Gly Ile Asn Asn Gly Asp Lys Asn Leu Val Glu Leu 
1               5                   10              


<210>  28
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 36 (v36)

<400>  28

Arg Gly Asp Asn Asn Pro Arg Gln Asn Met Ile Glu Val 
1               5                   10              


<210>  29
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 37 (v37)

<400>  29

Ile Gly Val Asn Asn Asp Ser Thr Asn Arg Val Glu Leu 
1               5                   10              


<210>  30
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 38 (v38)

<400>  30

Arg Gly Asp Asn Asp Pro Arg Arg Ser Thr Ile Glu Leu 
1               5                   10              


<210>  31
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 39 (v39)

<400>  31

Arg Gly Asp Asn Asn Pro Arg Gln Asn Lys Leu Glu Val 
1               5                   10              


<210>  32
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 40 (v40)

<400>  32

Thr Ala Thr Asn Asn Asp Lys Lys Asn Met Ile Glu Val 
1               5                   10              


<210>  33
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 42 (v42)

<400>  33

Ile Gly Val Asn Asn Asn Arg Leu Asn Lys Ile Glu Leu 
1               5                   10              


<210>  34
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 43 (v43)

<400>  34

Ile Gly Val Phe Ser Asp Ala Gly Asn Leu Leu Glu Val 
1               5                   10              


<210>  35
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 44 (v44)

<400>  35

Ile Gly Asp Asn Asn Pro Arg Asn Asn Val Ile Glu Val 
1               5                   10              


<210>  36
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 21-L (v21-L)

<400>  36

Ile Gly Val Asn Ser Asp Asp Arg Asn Leu Ile Glu Leu 
1               5                   10              


<210>  37
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 21-K (v21-K)

<400>  37

Ile Gly Val Asn Ser Asp Asp Arg Asn Lys Ile Glu Leu 
1               5                   10              


<210>  38
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of Cas 9 variant 21-R (v21-R)

<400>  38

Ile Gly Val Asn Ser Asp Asp Arg Asn Arg Ile Glu Leu 
1               5                   10              


<210>  39
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of SaCas9-N986S (N986S)

<400>  39

Ile Gly Val Asn Ser Asp Leu Leu Asn Arg Ile Glu Val 
1               5                   10              


<210>  40
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of SaCas9 KKH-D987N and R991K

<400>  40

Ile Gly Val Asn Asn Asn Leu Leu Asn Lys Ile Glu Val 
1               5                   10              


<210>  41
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of SaCas9 KKH-R991K

<400>  41

Ile Gly Val Asn Asn Asp Leu Leu Asn Lys Ile Glu Val 
1               5                   10              


<210>  42
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of  Cas9 variant 17-I991K

<400>  42

Ile Gly Val Asn Asn Ser Thr Arg Asn  Lys Val Glu Leu 
1               5                   10              


<210>  43
<211>  13
<212>  PRT
<213>  Artificial

<220>
<223>  PAM interaction region of  Cas9 variant 17-I991L
<400>  43

Ile Gly Val Asn Asn Ser Thr Arg Asn Leu Val Glu Leu 
1               5                   10              


<210>  44
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 10 (v10)

<400>  44

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Ser Asp Ser Ser Pro Arg Glu Asn Arg Leu 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  45
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 12 (v12)

<400>  45

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Lys Gly Asp Ala Met Pro Arg Gly Asn Lys Ile 
            980                 985                 990         


Glu Ile Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  46
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 13 (v13)

<400>  46

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Thr Ala Thr Asn Asn Asp Lys Ser Asn Lys Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  47
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 14 (v14)

<400>  47

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Ala Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  48
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 15 (v15)

<400>  48

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn His Asp Glu Gly Asn Arg Ile 
            980                 985                 990         


Glu Met Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  49
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 16 (v16)

<400>  49

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  50
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 17 (v17)

<400>  50

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Ser Thr Arg Asn Ile Val 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  51
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 18 (v18)

<400>  51

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Asn Asn Pro Arg Gln Asn Lys Leu 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  52
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 19 (v19)

<400>  52

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Lys Asn Asn Val Ile 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  53
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 20 (v20)

<400>  53

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Ile Asn Asp Asn Lys His Asn Lys Ile 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  54
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 21 (v21)

<400>  54

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  55
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 22 (v22)

<400>  55

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Gln Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  56
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 23 (v23)

<400>  56

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Lys Cys Ile Asn Asn Glu Lys Thr His Arg Ile 
            980                 985                 990         


Glu Ile Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  57
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 24 (v24)

<400>  57

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn His Asp Lys Thr Asn Arg Ile 
            980                 985                 990         


Glu Cys Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  58
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 25 (v25)

<400>  58

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Glu Asp Lys Arg Asn Ile Ile 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  59
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 26 (v26)

<400>  59

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Ser Ser Pro Arg Glu Asn Arg Phe 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  60
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant  27 (v27)

<400>  60

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Asn Asp Pro Lys Ala Asn Lys Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  61
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 28 (v28)

<400>  61

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Ala Glu Lys Arg Asn Thr Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  62
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 29 (v29)

<400>  62

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asp Asp Ala Lys Asn Thr Leu 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  63
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 30 (v30)

<400>  63

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Val Gly Val Asn Asn Asp Ser Val Asn Arg Val 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  64
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 31 (v31)

<400>  64

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Val Gly Val Asn Asn Asp Thr Arg Asn Val Val 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  65
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 32 (v32)

<400>  65

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Val Gly Val Asn Asn Asp Ser Arg Asn Val Val 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  66
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 33 (v33)

<400>  66

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Ser Met Pro Arg Gln Asn Lys Ile 
            980                 985                 990         


Glu Met Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  67
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 34 (v34)

<400>  67

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Ala Met Pro Arg Asp Asn Lys Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  68
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 35 (v35)

<400>  68

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Ile Asn Asn Gly Asp Lys Asn Leu Val 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  69
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 36 (v36)

<400>  69

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Asn Asn Pro Arg Gln Asn Met Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  70
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 37 (v37)

<400>  70

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  71
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 38 (v38)

<400>  71

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Asn Asp Pro Arg Arg Ser Thr Ile 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  72
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 39 (v39)

<400>  72

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Arg Gly Asp Asn Asn Pro Arg Gln Asn Lys Leu 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  73
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 40 (v40)

<400>  73

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Thr Ala Thr Asn Asn Asp Lys Lys Asn Met Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  74
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 42 (v42)

<400>  74

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asn Arg Leu Asn Lys Ile 
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  75
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 43 (v43)

<400>  75

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Phe Ser Asp Ala Gly Asn Leu Leu 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  76
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 44 (v44)

<400>  76

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Asp Asn Asn Pro Arg Asn Asn Val Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  77
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 21-L (v21-L)

<400>  77

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  78
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 21-K (v21-K)

<400>  78

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  79
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 21-R (v21-R)

<400>  79

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  80
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of SaCas9-N986S (N986S)

<400>  80

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  81
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of SaCas9 KKH-D987N and R991K

<400>  81

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asn Leu Leu Asn Lys Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  82
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of SaCas9 KKH-R991K

<400>  82

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Lys Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  83
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 17-I991K

<400>  83

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Ser Thr Arg Asn Lys Val
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  84
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant 17-I991L

<400>  84

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Ser Thr Arg Asn Leu Val
            980                 985                 990         


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  85
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 10 (v10)

<400>  85
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggagcg acagcagccc cagggagaac aggctggagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  86
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 12 (v12)

<400>  86
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaagggcg acgccatgcc caggggcaac aagatcgaga tctgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  87
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 13 (v13)

<400>  87
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaccgcca ccaacaacga caagagcaac aagatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  88
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 14 (v14)

<400>  88
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgctgggcg acgccaacag caggcagaac atcctggagg cctgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  89
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 15 (v15)

<400>  89
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaaccacga cgagggcaac aggatcgaga tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  90
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 16 (v16)

<400>  90
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacagcga caagaacaac ctgatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  91
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 17 (v17)

<400>  91
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacag caccaggaac atcgtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  92
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 18 (v18)

<400>  92
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggggcg acaacaaccc caggcagaac aagctggagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  93
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 19 (v19)

<400>  93
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacga caagaacaac gtgatcgagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  94
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 20 (v20) 

<400>  94
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggca tcaacgacaa caagcacaac aagatcgagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  95
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 21 (v21)

<400>  95
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacagcga cgacaggaac atcatcgagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  96
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 22 (v22)

<400>  96
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacgacag cgagaagaac aagatccagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  97
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 23 (v23)

<400>  97
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaagtgca tcaacaacga gaagacccac aggatcgaga tctgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  98
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 24 (v24)

<400>  98
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaaccacga caagaccaac aggatcgagt gctgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  99
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 25 (v25)

<400>  99
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc       60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac      120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg      180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac      240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg      300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac      360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg      420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa      480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc      540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc      600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc      660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc      720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac      780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag      840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc      900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag      960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag     1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc     1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc     1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc     1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg     1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg     1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg     1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc     1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag     1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg     1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc     1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc     1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc     1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc     1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag     1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac     1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg     1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc     2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac     2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa     2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc     2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc     2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat     2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg     2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc     2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg     2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa     2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt     2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc     2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat     2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac     2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc     2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga     2940

gtgatcggcg tgaacgagga caagaggaac atcatcgagc tgtgagacgg gccatactcg     3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc     3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag     3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg     3180

gc                                                                    3182


<210>  100
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 26 (v26)

<400>  100
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggggcg acagcagccc cagggagaac aggttcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  101
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 27 (v27)

<400>  101
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggggcg acaacgaccc caaggccaac aagatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  102
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 28 (v28)

<400>  102
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacgccga gaagaggaac accatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  103
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 29 (v29)

<400>  103
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacgacga cgccaagaac accctggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  104
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 30 (v30)

<400>  104
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtggtgggcg tgaacaacga cagcgtgaac agggtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  105
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 31 (v31)

<400>  105
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtggtgggcg tgaacaacga caccaggaac gtggtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  106
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 32 (v32)

<400>  106
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtggtgggcg tgaacaacga cagcaggaac gtggtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  107
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 33 (v33)

<400>  107
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggggcg acagcatgcc caggcagaac aagatcgaga tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  108
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 34 (v34)

<400>  108
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggggcg acgccatgcc cagggacaac aagatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  109
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 35 (v35)

<400>  109
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggca tcaacaacgg cgacaagaac ctggtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  110
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 36 (v36)

<400>  110
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggggcg acaacaaccc caggcagaac atgatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  111
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 37 (v37)

<400>  111
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacga cagcaccaac agggtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  112
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 38 (v38)

<400>  112
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc       60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac      120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg      180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac      240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg      300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac      360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg      420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa      480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc      540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc      600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc      660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc      720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac      780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag      840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc      900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag      960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag     1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc     1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc     1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc     1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg     1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg     1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg     1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc     1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag     1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg     1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc     1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc     1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc     1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc     1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag     1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac     1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg     1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc     2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac     2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa     2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc     2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc     2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat     2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg     2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc     2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg     2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa     2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt     2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc     2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat     2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac     2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc     2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga     2940

gtgaggggcg acaacgaccc caggaggagc accatcgagc tgtgagacgg gccatactcg     3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc     3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag     3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg     3180

gc                                                                    3182


<210>  113
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 39 (v39)

<400>  113
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaggggcg acaacaaccc caggcagaac aagctggagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  114
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 40 (v40)

<400>  114
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgaccgcca ccaacaacga caagaagaac atgatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  115
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 42 (v42)

<400>  115
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacaa caggctgaac aagatcgagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  116
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 43 (v43)

<400>  116
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgttcagcga cgccggcaac ctgctggagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  117
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 44 (v44)

<400>  117
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg acaacaaccc caggaacaac gtgatcgagg tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  118
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding SaCas9 KKH-D987N and R991K

<400>  118
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacaa cctgctgaac aagatcgaag tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  119
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding SaCas9-N986S (N986S)

<400>  119
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaactcgga cctgctgaac cggatcgaag tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  120
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding SaCas9 KKH-R991K

<400>  120
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacga cctgctgaac aagatcgaag tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  121
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 17-I991K

<400>  121
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacag caccaggaac aaggtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  122
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 17-I991L

<400>  122
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacaacag caccaggaac ctggtggagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  123
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 21-K (v21-K)

<400>  123
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacagcga cgacaggaac aagatcgagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  124
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 21-L (v21-L)

<400>  124
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacagcga cgacaggaac ctgatcgagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  125
<211>  3182
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant 21-R (v21-R)

<400>  125
atgaagcgga actacatcct gggcctggcc atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaagcc   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacagcga cgacaggaac aggatcgagc tgtgagacgg gccatactcg   3000

tctcgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc   3060

ccccccacat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag   3120

acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg   3180

gc                                                                  3182


<210>  126
<211>  82
<212>  DNA
<213>  Artificial

<220>
<223>  second nucleic acid sequence encoding scaffold structure of wild-type gRNA

<400>  126
gttttagtac tctggaaaca gaatctacta aaacaaggca aaatgccgtg tttatctcgt     60

caacttgttg gcgagatttt tt                                              82


<210>  127
<211>  82
<212>  DNA
<213>  Artificial

<220>
<223>  second nucleic acid sequence encoding scaffold structure of gRNA2

<400>  127
gtcttagtac tctggaaaca gaatctacta agacaaggca aaatgccgtg tttatctcgt     60

caacttgttg gcgagatttt tt                                              82


<210>  128
<211>  83
<212>  DNA
<213>  Artificial

<220>
<223>  second nucleic acid sequence encoding scaffold structure of gRNA1

<400>  128
gttatagtac tctggaaaca gaatctacta taacaaggca aaatgccgtg tttatctcgt     60

caacttgttg gcgagatttt ttt                                             83


<210>  129
<211>  83
<212>  DNA
<213>  Artificial

<220>
<223>  second nucleic acid sequence encoding scaffold structure of gRNA3

<400>  129
gttccggtac tctggaaaca gaatctaccg gaacaaggca aaatgccgtg tttatctcgt     60

caacttgttg gcgagatttt ttt                                             83


<210>  130
<211>  981
<212>  PRT
<213>  Artificial

<220>
<223>  first backbone region of  Cas9 variant 21-R-HF

<400>  130

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Ala Lys Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Ala Arg Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val 
            980     


<210>  131
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of Cas9 variant v21-R-HF

<400>  131

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Ala Lys Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Ala Arg Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Lys Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Lys Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


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


Glu Leu Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro His  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  132
<211>  3159
<212>  DNA
<213>  Artificial

<220>
<223>  nucleotide sequence of nucleic acid encoding Cas9 variant v21-R-HF

<400>  132
atgaagcgga actacatcct gggcctggac atcggcatca ccagcgtggg ctacggcatc     60

atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac    120

gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg    180

cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac    240

agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg    300

agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac    360

gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg    420

aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa    480

gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc    540

aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc    600

tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc    660

ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc    720

cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac    780

gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag    840

ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc    900

aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag    960

cccgagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag   1020

attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc   1080

agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc   1140

gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc   1200

aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg   1260

ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg   1320

gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg   1380

atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc   1440

gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaacgccaag   1500

accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg   1560

atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc   1620

atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc   1680

agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaaaac   1740

agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc   1800

tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag   1860

accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac   1920

ttcatcaacc ggaacctggt ggataccaga tacgccaccg cccggctgat gaacctgctg   1980

cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc   2040

accagctttc tgcggcggaa gtggaagttt aagaaagagc ggaacaaggg gtacaagcac   2100

cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa   2160

ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc   2220

atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc   2280

aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat   2340

agaaagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg   2400

atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc   2460

aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg   2520

aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa   2580

accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt   2640

aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc   2700

agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat   2760

ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac   2820

gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc   2880

gagtttatcg cctccttcta caagaacgat ctgatcaaga tcaacggcga gctgtataga   2940

gtgatcggcg tgaacagcga cgaccggaac cggatcgaag tgctgatgat cgacatcacc   3000

taccgcgagt acctggaaaa catgaacgac aagaggcccc cccacatcat taagacaatc   3060

gcctccaaga cccagagcat taagaagtac agcacagaca ttctgggcaa cctgtatgaa   3120

gtgaaatcta agaagcaccc tcagatcatc aaaaagggc                          3159


<210>  133
<211>  1053
<212>  PRT
<213>  Artificial

<220>
<223>  amino acid sequence of wild-type SaCas9

<400>  133

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


Gly Tyr Gly Ile Ile Asp Tyr Glu Thr Arg Asp Val Ile Asp Ala Gly 
            20                  25                  30          


Val Arg Leu Phe Lys Glu Ala Asn Val Glu Asn Asn Glu Gly Arg Arg 
        35                  40                  45              


Ser Lys Arg Gly Ala Arg Arg Leu Lys Arg Arg Arg Arg His Arg Ile 
    50                  55                  60                  


Gln Arg Val Lys Lys Leu Leu Phe Asp Tyr Asn Leu Leu Thr Asp His 
65                  70                  75                  80  


Ser Glu Leu Ser Gly Ile Asn Pro Tyr Glu Ala Arg Val Lys Gly Leu 
                85                  90                  95      


Ser Gln Lys Leu Ser Glu Glu Glu Phe Ser Ala Ala Leu Leu His Leu 
            100                 105                 110         


Ala Lys Arg Arg Gly Val His Asn Val Asn Glu Val Glu Glu Asp Thr 
        115                 120                 125             


Gly Asn Glu Leu Ser Thr Lys Glu Gln Ile Ser Arg Asn Ser Lys Ala 
    130                 135                 140                 


Leu Glu Glu Lys Tyr Val Ala Glu Leu Gln Leu Glu Arg Leu Lys Lys 
145                 150                 155                 160 


Asp Gly Glu Val Arg Gly Ser Ile Asn Arg Phe Lys Thr Ser Asp Tyr 
                165                 170                 175     


Val Lys Glu Ala Lys Gln Leu Leu Lys Val Gln Lys Ala Tyr His Gln 
            180                 185                 190         


Leu Asp Gln Ser Phe Ile Asp Thr Tyr Ile Asp Leu Leu Glu Thr Arg 
        195                 200                 205             


Arg Thr Tyr Tyr Glu Gly Pro Gly Glu Gly Ser Pro Phe Gly Trp Lys 
    210                 215                 220                 


Asp Ile Lys Glu Trp Tyr Glu Met Leu Met Gly His Cys Thr Tyr Phe 
225                 230                 235                 240 


Pro Glu Glu Leu Arg Ser Val Lys Tyr Ala Tyr Asn Ala Asp Leu Tyr 
                245                 250                 255     


Asn Ala Leu Asn Asp Leu Asn Asn Leu Val Ile Thr Arg Asp Glu Asn 
            260                 265                 270         


Glu Lys Leu Glu Tyr Tyr Glu Lys Phe Gln Ile Ile Glu Asn Val Phe 
        275                 280                 285             


Lys Gln Lys Lys Lys Pro Thr Leu Lys Gln Ile Ala Lys Glu Ile Leu 
    290                 295                 300                 


Val Asn Glu Glu Asp Ile Lys Gly Tyr Arg Val Thr Ser Thr Gly Lys 
305                 310                 315                 320 


Pro Glu Phe Thr Asn Leu Lys Val Tyr His Asp Ile Lys Asp Ile Thr 
                325                 330                 335     


Ala Arg Lys Glu Ile Ile Glu Asn Ala Glu Leu Leu Asp Gln Ile Ala 
            340                 345                 350         


Lys Ile Leu Thr Ile Tyr Gln Ser Ser Glu Asp Ile Gln Glu Glu Leu 
        355                 360                 365             


Thr Asn Leu Asn Ser Glu Leu Thr Gln Glu Glu Ile Glu Gln Ile Ser 
    370                 375                 380                 


Asn Leu Lys Gly Tyr Thr Gly Thr His Asn Leu Ser Leu Lys Ala Ile 
385                 390                 395                 400 


Asn Leu Ile Leu Asp Glu Leu Trp His Thr Asn Asp Asn Gln Ile Ala 
                405                 410                 415     


Ile Phe Asn Arg Leu Lys Leu Val Pro Lys Lys Val Asp Leu Ser Gln 
            420                 425                 430         


Gln Lys Glu Ile Pro Thr Thr Leu Val Asp Asp Phe Ile Leu Ser Pro 
        435                 440                 445             


Val Val Lys Arg Ser Phe Ile Gln Ser Ile Lys Val Ile Asn Ala Ile 
    450                 455                 460                 


Ile Lys Lys Tyr Gly Leu Pro Asn Asp Ile Ile Ile Glu Leu Ala Arg 
465                 470                 475                 480 


Glu Lys Asn Ser Lys Asp Ala Gln Lys Met Ile Asn Glu Met Gln Lys 
                485                 490                 495     


Arg Asn Arg Gln Thr Asn Glu Arg Ile Glu Glu Ile Ile Arg Thr Thr 
            500                 505                 510         


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


Met Gln Glu Gly Lys Cys Leu Tyr Ser Leu Glu Ala Ile Pro Leu Glu 
    530                 535                 540                 


Asp Leu Leu Asn Asn Pro Phe Asn Tyr Glu Val Asp His Ile Ile Pro 
545                 550                 555                 560 


Arg Ser Val Ser Phe Asp Asn Ser Phe Asn Asn Lys Val Leu Val Lys 
                565                 570                 575     


Gln Glu Glu Asn Ser Lys Lys Gly Asn Arg Thr Pro Phe Gln Tyr Leu 
            580                 585                 590         


Ser Ser Ser Asp Ser Lys Ile Ser Tyr Glu Thr Phe Lys Lys His Ile 
        595                 600                 605             


Leu Asn Leu Ala Lys Gly Lys Gly Arg Ile Ser Lys Thr Lys Lys Glu 
    610                 615                 620                 


Tyr Leu Leu Glu Glu Arg Asp Ile Asn Arg Phe Ser Val Gln Lys Asp 
625                 630                 635                 640 


Phe Ile Asn Arg Asn Leu Val Asp Thr Arg Tyr Ala Thr Arg Gly Leu 
                645                 650                 655     


Met Asn Leu Leu Arg Ser Tyr Phe Arg Val Asn Asn Leu Asp Val Lys 
            660                 665                 670         


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


Lys Phe Lys Lys Glu Arg Asn Lys Gly Tyr Lys His His Ala Glu Asp 
    690                 695                 700                 


Ala Leu Ile Ile Ala Asn Ala Asp Phe Ile Phe Lys Glu Trp Lys Lys 
705                 710                 715                 720 


Leu Asp Lys Ala Lys Lys Val Met Glu Asn Gln Met Phe Glu Glu Lys 
                725                 730                 735     


Gln Ala Glu Ser Met Pro Glu Ile Glu Thr Glu Gln Glu Tyr Lys Glu 
            740                 745                 750         


Ile Phe Ile Thr Pro His Gln Ile Lys His Ile Lys Asp Phe Lys Asp 
        755                 760                 765             


Tyr Lys Tyr Ser His Arg Val Asp Lys Lys Pro Asn Arg Glu Leu Ile 
    770                 775                 780                 


Asn Asp Thr Leu Tyr Ser Thr Arg Lys Asp Asp Lys Gly Asn Thr Leu 
785                 790                 795                 800 


Ile Val Asn Asn Leu Asn Gly Leu Tyr Asp Lys Asp Asn Asp Lys Leu 
                805                 810                 815     


Lys Lys Leu Ile Asn Lys Ser Pro Glu Lys Leu Leu Met Tyr His His 
            820                 825                 830         


Asp Pro Gln Thr Tyr Gln Lys Leu Lys Leu Ile Met Glu Gln Tyr Gly 
        835                 840                 845             


Asp Glu Lys Asn Pro Leu Tyr Lys Tyr Tyr Glu Glu Thr Gly Asn Tyr 
    850                 855                 860                 


Leu Thr Lys Tyr Ser Lys Lys Asp Asn Gly Pro Val Ile Lys Lys Ile 
865                 870                 875                 880 


Lys Tyr Tyr Gly Asn Lys Leu Asn Ala His Leu Asp Ile Thr Asp Asp 
                885                 890                 895     


Tyr Pro Asn Ser Arg Asn Lys Val Val Lys Leu Ser Leu Lys Pro Tyr 
            900                 905                 910         


Arg Phe Asp Val Tyr Leu Asp Asn Gly Val Tyr Lys Phe Val Thr Val 
        915                 920                 925             


Lys Asn Leu Asp Val Ile Lys Lys Glu Asn Tyr Tyr Glu Val Asn Ser 
    930                 935                 940                 


Lys Cys Tyr Glu Glu Ala Lys Lys Leu Lys Lys Ile Ser Asn Gln Ala 
945                 950                 955                 960 


Glu Phe Ile Ala Ser Phe Tyr Asn Asn Asp Leu Ile Lys Ile Asn Gly 
                965                 970                 975     


Glu Leu Tyr Arg Val Ile Gly Val Asn Asn Asp Leu Leu Asn Arg Ile 
            980                 985                 990         


Glu Val Asn Met Ile Asp Ile Thr  Tyr Arg Glu Tyr Leu  Glu Asn Met 
        995                 1000                 1005             


Asn Asp  Lys Arg Pro Pro Arg  Ile Ile Lys Thr Ile  Ala Ser Lys 
    1010                 1015                 1020             


Thr Gln  Ser Ile Lys Lys Tyr  Ser Thr Asp Ile Leu  Gly Asn Leu 
    1025                 1030                 1035             


Tyr Glu  Val Lys Ser Lys Lys  His Pro Gln Ile Ile  Lys Lys Gly 
    1040                 1045                 1050             


<210>  134
<211>  21
<212>  DNA
<213>  Artificial

<220>
<223>  -actin-F primer

<400>  134
catgtacgtt gctatccagg c                                                 21


<210>  135
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  -actin-R primer

<400>  135
ctccttaatg tcacgcacga t                                                 21


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

<220>
<223>  IL1RN-F primer

<400>  136
ggaatccatg gagggaagat                                                   20


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

<220>
<223>  IL1RN-R primer

<400>  137
tgttctcgct caggtcagtg                                                   20
