                         SEQUENCE LISTING

<110>  THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK
 
<120>  ENGINEERED Cas-Transposon SYSTEM FOR PROGRAMMABLE AND 
       SITE-DIRECTED DNA TRANSPOSITIONS

<130>  15003-443PC0

<140>  PCT/US2020/034538
<141>  2020-05-26

<160>  52    

<170>  PatentIn version 3.5

<210>  1
<211>  348
<212>  PRT
<213>  Haematobia irritans


<220>
<221>  misc_feature
<223>  Himar1 WT

<400>  1

Met Glu Lys Lys Glu Phe Arg Val Leu Ile Lys Tyr Cys Phe Leu Lys 
1               5                   10                  15      


Gly Lys Asn Thr Val Glu Ala Lys Thr Trp Leu Asp Asn Glu Phe Pro 
            20                  25                  30          


Asp Ser Ala Pro Gly Lys Ser Thr Ile Ile Asp Trp Tyr Ala Lys Phe 
        35                  40                  45              


Lys Arg Gly Glu Met Ser Thr Glu Asp Gly Glu Arg Ser Gly Arg Pro 
    50                  55                  60                  


Lys Glu Val Val Thr Asp Glu Asn Ile Lys Lys Ile His Lys Met Ile 
65                  70                  75                  80  


Leu Asn Asp Arg Lys Met Lys Leu Ile Glu Ile Ala Glu Ala Leu Lys 
                85                  90                  95      


Ile Ser Lys Glu Arg Val Gly His Ile Ile His Gln Tyr Leu Asp Met 
            100                 105                 110         


Arg Lys Leu Cys Ala Lys Trp Val Pro Arg Glu Leu Thr Phe Asp Gln 
        115                 120                 125             


Lys Gln Gln Arg Val Asp Asp Ser Glu Arg Cys Leu Gln Leu Leu Thr 
    130                 135                 140                 


Arg Asn Thr Pro Glu Phe Phe Arg Arg Tyr Val Thr Met Asp Glu Thr 
145                 150                 155                 160 


Trp Leu His His Tyr Thr Pro Glu Ser Asn Arg Gln Ser Ala Glu Trp 
                165                 170                 175     


Thr Ala Thr Gly Glu Pro Ser Pro Lys Arg Gly Lys Thr Gln Lys Ser 
            180                 185                 190         


Ala Gly Lys Val Met Ala Ser Val Phe Trp Asp Ala His Gly Ile Ile 
        195                 200                 205             


Phe Ile Asp Tyr Leu Glu Lys Gly Lys Thr Ile Asn Ser Asp Tyr Tyr 
    210                 215                 220                 


Met Ala Leu Leu Glu Arg Leu Lys Val Glu Ile Ala Ala Lys Arg Pro 
225                 230                 235                 240 


His Met Lys Lys Lys Lys Val Leu Phe His Gln Asp Asn Ala Pro Cys 
                245                 250                 255     


His Lys Ser Leu Arg Thr Met Ala Lys Ile His Glu Leu Gly Phe Glu 
            260                 265                 270         


Leu Leu Pro His Pro Pro Tyr Ser Pro Asp Leu Ala Pro Ser Asp Phe 
        275                 280                 285             


Phe Leu Phe Ser Asp Leu Lys Arg Met Leu Ala Gly Lys Lys Phe Gly 
    290                 295                 300                 


Cys Asn Glu Glu Val Ile Ala Glu Thr Glu Ala Tyr Phe Glu Ala Lys 
305                 310                 315                 320 


Pro Lys Glu Tyr Tyr Gln Asn Gly Ile Lys Lys Leu Glu Gly Arg Tyr 
                325                 330                 335     


Asn Arg Cys Ile Ala Leu Glu Gly Asn Tyr Val Glu 
            340                 345             


<210>  2
<211>  348
<212>  PRT
<213>  Haematobia irritans


<220>
<221>  misc_feature
<223>  Himar1C9

<400>  2

Met Glu Lys Lys Glu Phe Arg Val Leu Ile Lys Tyr Cys Phe Leu Lys 
1               5                   10                  15      


Gly Lys Asn Thr Val Glu Ala Lys Thr Trp Leu Asp Asn Glu Phe Pro 
            20                  25                  30          


Asp Ser Ala Pro Gly Lys Ser Thr Ile Ile Asp Trp Tyr Ala Lys Phe 
        35                  40                  45              


Lys Arg Gly Glu Met Ser Thr Glu Asp Gly Glu Arg Ser Gly Arg Pro 
    50                  55                  60                  


Lys Glu Val Val Thr Asp Glu Asn Ile Lys Lys Ile His Lys Met Ile 
65                  70                  75                  80  


Leu Asn Asp Arg Lys Met Lys Leu Ile Glu Ile Ala Glu Ala Leu Lys 
                85                  90                  95      


Ile Ser Lys Glu Arg Val Gly His Ile Ile His Gln Tyr Leu Asp Met 
            100                 105                 110         


Arg Lys Leu Cys Ala Lys Trp Val Pro Arg Glu Leu Thr Phe Asp Gln 
        115                 120                 125             


Lys Gln Arg Arg Val Asp Asp Ser Lys Arg Cys Leu Gln Leu Leu Thr 
    130                 135                 140                 


Arg Asn Thr Pro Glu Phe Phe Arg Arg Tyr Val Thr Met Asp Glu Thr 
145                 150                 155                 160 


Trp Leu His His Tyr Thr Pro Glu Ser Asn Arg Gln Ser Ala Glu Trp 
                165                 170                 175     


Thr Ala Thr Gly Glu Pro Ser Pro Lys Arg Gly Lys Thr Gln Lys Ser 
            180                 185                 190         


Ala Gly Lys Val Met Ala Ser Val Phe Trp Asp Ala His Gly Ile Ile 
        195                 200                 205             


Phe Ile Asp Tyr Leu Glu Lys Gly Lys Thr Ile Asn Ser Asp Tyr Tyr 
    210                 215                 220                 


Met Ala Leu Leu Glu Arg Leu Lys Val Glu Ile Ala Ala Lys Arg Pro 
225                 230                 235                 240 


His Met Lys Lys Lys Lys Val Leu Phe His Gln Asp Asn Ala Pro Cys 
                245                 250                 255     


His Lys Ser Leu Arg Thr Met Ala Lys Ile His Glu Leu Gly Phe Glu 
            260                 265                 270         


Leu Leu Pro His Pro Pro Tyr Ser Pro Asp Leu Ala Pro Ser Asp Phe 
        275                 280                 285             


Phe Leu Phe Ser Asp Leu Lys Arg Met Leu Ala Gly Lys Lys Phe Gly 
    290                 295                 300                 


Cys Asn Glu Glu Val Ile Ala Glu Thr Glu Ala Tyr Phe Glu Ala Lys 
305                 310                 315                 320 


Pro Lys Glu Tyr Tyr Gln Asn Gly Ile Lys Lys Leu Glu Gly Arg Tyr 
                325                 330                 335     


Asn Arg Cys Ile Ala Leu Glu Gly Asn Tyr Val Glu 
            340                 345             


<210>  3
<211>  1732
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Himar1C9-dCas9 fusion protein

<400>  3

Met Glu Lys Lys Glu Phe Arg Val Leu Ile Lys Tyr Cys Phe Leu Lys 
1               5                   10                  15      


Gly Lys Asn Thr Val Glu Ala Lys Thr Trp Leu Asp Asn Glu Phe Pro 
            20                  25                  30          


Asp Ser Ala Pro Gly Lys Ser Thr Ile Ile Asp Trp Tyr Ala Lys Phe 
        35                  40                  45              


Lys Arg Gly Glu Met Ser Thr Glu Asp Gly Glu Arg Ser Gly Arg Pro 
    50                  55                  60                  


Lys Glu Val Val Thr Asp Glu Asn Ile Lys Lys Ile His Lys Met Ile 
65                  70                  75                  80  


Leu Asn Asp Arg Lys Met Lys Leu Ile Glu Ile Ala Glu Ala Leu Lys 
                85                  90                  95      


Ile Ser Lys Glu Arg Val Gly His Ile Ile His Gln Tyr Leu Asp Met 
            100                 105                 110         


Arg Lys Leu Cys Ala Lys Trp Val Pro Arg Glu Leu Thr Phe Asp Gln 
        115                 120                 125             


Lys Gln Arg Arg Val Asp Asp Ser Lys Arg Cys Leu Gln Leu Leu Thr 
    130                 135                 140                 


Arg Asn Thr Pro Glu Phe Phe Arg Arg Tyr Val Thr Met Asp Glu Thr 
145                 150                 155                 160 


Trp Leu His His Tyr Thr Pro Glu Ser Asn Arg Gln Ser Ala Glu Trp 
                165                 170                 175     


Thr Ala Thr Gly Glu Pro Ser Pro Lys Arg Gly Lys Thr Gln Lys Ser 
            180                 185                 190         


Ala Gly Lys Val Met Ala Ser Val Phe Trp Asp Ala His Gly Ile Ile 
        195                 200                 205             


Phe Ile Asp Tyr Leu Glu Lys Gly Lys Thr Ile Asn Ser Asp Tyr Tyr 
    210                 215                 220                 


Met Ala Leu Leu Glu Arg Leu Lys Val Glu Ile Ala Ala Lys Arg Pro 
225                 230                 235                 240 


His Met Lys Lys Lys Lys Val Leu Phe His Gln Asp Asn Ala Pro Cys 
                245                 250                 255     


His Lys Ser Leu Arg Thr Met Ala Lys Ile His Glu Leu Gly Phe Glu 
            260                 265                 270         


Leu Leu Pro His Pro Pro Tyr Ser Pro Asp Leu Ala Pro Ser Asp Phe 
        275                 280                 285             


Phe Leu Phe Ser Asp Leu Lys Arg Met Leu Ala Gly Lys Lys Phe Gly 
    290                 295                 300                 


Cys Asn Glu Glu Val Ile Ala Glu Thr Glu Ala Tyr Phe Glu Ala Lys 
305                 310                 315                 320 


Pro Lys Glu Tyr Tyr Gln Asn Gly Ile Lys Lys Leu Glu Gly Arg Tyr 
                325                 330                 335     


Asn Arg Cys Ile Ala Leu Glu Gly Asn Tyr Val Glu Ser Gly Ser Glu 
            340                 345                 350         


Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Met Asp Lys Lys 
        355                 360                 365             


Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val 
    370                 375                 380                 


Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly 
385                 390                 395                 400 


Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu 
                405                 410                 415     


Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala 
            420                 425                 430         


Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu 
        435                 440                 445             


Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe His Arg 
    450                 455                 460                 


Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu Arg His 
465                 470                 475                 480 


Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr 
                485                 490                 495     


Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys 
            500                 505                 510         


Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe 
        515                 520                 525             


Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp 
    530                 535                 540                 


Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe 
545                 550                 555                 560 


Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu 
                565                 570                 575     


Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln 
            580                 585                 590         


Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu 
        595                 600                 605             


Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu 
    610                 615                 620                 


Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp 
625                 630                 635                 640 


Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala 
                645                 650                 655     


Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val 
            660                 665                 670         


Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg 
        675                 680                 685             


Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg 
    690                 695                 700                 


Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys 
705                 710                 715                 720 


Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe 
                725                 730                 735     


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu 
            740                 745                 750         


Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr 
        755                 760                 765             


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu His 
    770                 775                 780                 


Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn 
785                 790                 795                 800 


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
                805                 810                 815     


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys 
            820                 825                 830         


Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
        835                 840                 845             


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys 
    850                 855                 860                 


Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
865                 870                 875                 880 


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
                885                 890                 895     


Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
            900                 905                 910         


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
        915                 920                 925             


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
    930                 935                 940                 


Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
945                 950                 955                 960 


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
                965                 970                 975     


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
            980                 985                 990         


Arg Glu Met Ile Glu Glu Arg Leu  Lys Thr Tyr Ala His  Leu Phe Asp 
        995                 1000                 1005             


Asp Lys  Val Met Lys Gln Leu  Lys Arg Arg Arg Tyr  Thr Gly Trp 
    1010                 1015                 1020             


Gly Arg  Leu Ser Arg Lys Leu  Ile Asn Gly Ile Arg  Asp Lys Gln 
    1025                 1030                 1035             


Ser Gly  Lys Thr Ile Leu Asp  Phe Leu Lys Ser Asp  Gly Phe Ala 
    1040                 1045                 1050             


Asn Arg  Asn Phe Met Gln Leu  Ile His Asp Asp Ser  Leu Thr Phe 
    1055                 1060                 1065             


Lys Glu  Asp Ile Gln Lys Ala  Gln Val Ser Gly Gln  Gly Asp Ser 
    1070                 1075                 1080             


Leu His  Glu His Ile Ala Asn  Leu Ala Gly Ser Pro  Ala Ile Lys 
    1085                 1090                 1095             


Lys Gly  Ile Leu Gln Thr Val  Lys Val Val Asp Glu  Leu Val Lys 
    1100                 1105                 1110             


Val Met  Gly Arg His Lys Pro  Glu Asn Ile Val Ile  Glu Met Ala 
    1115                 1120                 1125             


Arg Glu  Asn Gln Thr Thr Gln  Lys Gly Gln Lys Asn  Ser Arg Glu 
    1130                 1135                 1140             


Arg Met  Lys Arg Ile Glu Glu  Gly Ile Lys Glu Leu  Gly Ser Gln 
    1145                 1150                 1155             


Ile Leu  Lys Glu His Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
    1160                 1165                 1170             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1175                 1180                 1185             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1190                 1195                 1200             


Ala Ile  Val Pro Gln Ser Phe  Leu Lys Asp Asp Ser  Ile Asp Asn 
    1205                 1210                 1215             


Lys Val  Leu Thr Arg Ser Asp  Lys Asn Arg Gly Lys  Ser Asp Asn 
    1220                 1225                 1230             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1235                 1240                 1245             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1250                 1255                 1260             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Leu  Asp Lys Ala 
    1265                 1270                 1275             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1280                 1285                 1290             


His Val  Ala Gln Ile Leu Asp  Ser Arg Met Asn Thr  Lys Tyr Asp 
    1295                 1300                 1305             


Glu Asn  Asp Lys Leu Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1310                 1315                 1320             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Phe Tyr Lys 
    1325                 1330                 1335             


Val Arg  Glu Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1340                 1345                 1350             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1355                 1360                 1365             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1370                 1375                 1380             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1385                 1390                 1395             


Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1400                 1405                 1410             


Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1415                 1420                 1425             


Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp Asp Lys  Gly Arg Asp 
    1430                 1435                 1440             


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


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1460                 1465                 1470             


Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu Ile Ala  Arg Lys Lys 
    1475                 1480                 1485             


Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe Asp Ser  Pro Thr Val 
    1490                 1495                 1500             


Ala Tyr  Ser Val Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ser 
    1505                 1510                 1515             


Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu Gly Ile  Thr Ile Met 
    1520                 1525                 1530             


Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile Asp Phe  Leu Glu Ala 
    1535                 1540                 1545             


Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu Ile Ile  Lys Leu Pro 
    1550                 1555                 1560             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Lys  Arg Met Leu 
    1565                 1570                 1575             


Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn Glu Leu  Ala Leu Pro 
    1580                 1585                 1590             


Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala Ser His  Tyr Glu Lys 
    1595                 1600                 1605             


Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln Lys Gln  Leu Phe Val 
    1610                 1615                 1620             


Glu Gln  His Lys His Tyr Leu  Asp Glu Ile Ile Glu  Gln Ile Ser 
    1625                 1630                 1635             


Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp Ala Asn  Leu Asp Lys 
    1640                 1645                 1650             


Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp Lys Pro  Ile Arg Glu 
    1655                 1660                 1665             


Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr Leu Thr  Asn Leu Gly 
    1670                 1675                 1680             


Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr Thr Ile  Asp Arg Lys 
    1685                 1690                 1695             


Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp Ala Thr  Leu Ile His 
    1700                 1705                 1710             


Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg Ile Asp  Leu Ser Gln 
    1715                 1720                 1725             


Leu Gly  Gly Asp 
    1730         


<210>  4
<211>  476
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Hyperactive Tn5 transposase

<400>  4

Met Ile Thr Ser Ala Leu His Arg Ala Ala Asp Trp Ala Lys Ser Val 
1               5                   10                  15      


Phe Ser Ser Ala Ala Leu Gly Asp Pro Arg Arg Thr Ala Arg Leu Val 
            20                  25                  30          


Asn Val Ala Ala Gln Leu Ala Lys Tyr Ser Gly Lys Ser Ile Thr Ile 
        35                  40                  45              


Ser Ser Glu Gly Ser Lys Ala Ala Gln Glu Gly Ala Tyr Arg Phe Ile 
    50                  55                  60                  


Arg Asn Pro Asn Val Ser Ala Glu Ala Ile Arg Lys Ala Gly Ala Met 
65                  70                  75                  80  


Gln Thr Val Lys Leu Ala Gln Glu Phe Pro Glu Leu Leu Ala Ile Glu 
                85                  90                  95      


Asp Thr Thr Ser Leu Ser Tyr Arg His Gln Val Ala Glu Glu Leu Gly 
            100                 105                 110         


Lys Leu Gly Ser Ile Gln Asp Lys Ser Arg Gly Trp Trp Val His Ser 
        115                 120                 125             


Val Leu Leu Leu Glu Ala Thr Thr Phe Arg Thr Val Gly Leu Leu His 
    130                 135                 140                 


Gln Glu Trp Trp Met Arg Pro Asp Asp Pro Ala Asp Ala Asp Glu Lys 
145                 150                 155                 160 


Glu Ser Gly Lys Trp Leu Ala Ala Ala Ala Thr Ser Arg Leu Arg Met 
                165                 170                 175     


Gly Ser Met Met Ser Asn Val Ile Ala Val Cys Asp Arg Glu Ala Asp 
            180                 185                 190         


Ile His Ala Tyr Leu Gln Asp Lys Leu Ala His Asn Glu Arg Phe Val 
        195                 200                 205             


Val Arg Ser Lys His Pro Arg Lys Asp Val Glu Ser Gly Leu Tyr Leu 
    210                 215                 220                 


Tyr Asp His Leu Lys Asn Gln Pro Glu Leu Gly Gly Tyr Gln Ile Ser 
225                 230                 235                 240 


Ile Pro Gln Lys Gly Val Val Asp Lys Arg Gly Lys Arg Lys Asn Arg 
                245                 250                 255     


Pro Ala Arg Lys Ala Ser Leu Ser Leu Arg Ser Gly Arg Ile Thr Leu 
            260                 265                 270         


Lys Gln Gly Asn Ile Thr Leu Asn Ala Val Leu Ala Glu Glu Ile Asn 
        275                 280                 285             


Pro Pro Lys Gly Glu Thr Pro Leu Lys Trp Leu Leu Leu Thr Ser Glu 
    290                 295                 300                 


Pro Val Glu Ser Leu Ala Gln Ala Leu Arg Val Ile Asp Ile Tyr Thr 
305                 310                 315                 320 


His Arg Trp Arg Ile Glu Glu Phe His Lys Ala Trp Lys Thr Gly Ala 
                325                 330                 335     


Gly Ala Glu Arg Gln Arg Met Glu Glu Pro Asp Asn Leu Glu Arg Met 
            340                 345                 350         


Val Ser Ile Leu Ser Phe Val Ala Val Arg Leu Leu Gln Leu Arg Glu 
        355                 360                 365             


Ser Phe Thr Pro Pro Gln Ala Leu Arg Ala Gln Gly Leu Leu Lys Glu 
    370                 375                 380                 


Ala Glu His Val Glu Ser Gln Ser Ala Glu Thr Val Leu Thr Pro Asp 
385                 390                 395                 400 


Glu Cys Gln Leu Leu Gly Tyr Leu Asp Lys Gly Lys Arg Lys Arg Lys 
                405                 410                 415     


Glu Lys Ala Gly Ser Leu Gln Trp Ala Tyr Met Ala Ile Ala Arg Leu 
            420                 425                 430         


Gly Gly Phe Met Asp Ser Lys Arg Thr Gly Ile Ala Ser Trp Gly Ala 
        435                 440                 445             


Leu Trp Glu Gly Trp Glu Ala Leu Gln Ser Lys Leu Asp Gly Phe Leu 
    450                 455                 460                 


Ala Ala Lys Asp Leu Met Ala Gln Gly Ile Lys Ile 
465                 470                 475     


<210>  5
<211>  1860
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Tn5-dCas9 fusion protein with XTEN linker

<400>  5

Met Ile Thr Ser Ala Leu His Arg Ala Ala Asp Trp Ala Lys Ser Val 
1               5                   10                  15      


Phe Ser Ser Ala Ala Leu Gly Asp Pro Arg Arg Thr Ala Arg Leu Val 
            20                  25                  30          


Asn Val Ala Ala Gln Leu Ala Lys Tyr Ser Gly Lys Ser Ile Thr Ile 
        35                  40                  45              


Ser Ser Glu Gly Ser Lys Ala Ala Gln Glu Gly Ala Tyr Arg Phe Ile 
    50                  55                  60                  


Arg Asn Pro Asn Val Ser Ala Glu Ala Ile Arg Lys Ala Gly Ala Met 
65                  70                  75                  80  


Gln Thr Val Lys Leu Ala Gln Glu Phe Pro Glu Leu Leu Ala Ile Glu 
                85                  90                  95      


Asp Thr Thr Ser Leu Ser Tyr Arg His Gln Val Ala Glu Glu Leu Gly 
            100                 105                 110         


Lys Leu Gly Ser Ile Gln Asp Lys Ser Arg Gly Trp Trp Val His Ser 
        115                 120                 125             


Val Leu Leu Leu Glu Ala Thr Thr Phe Arg Thr Val Gly Leu Leu His 
    130                 135                 140                 


Gln Glu Trp Trp Met Arg Pro Asp Asp Pro Ala Asp Ala Asp Glu Lys 
145                 150                 155                 160 


Glu Ser Gly Lys Trp Leu Ala Ala Ala Ala Thr Ser Arg Leu Arg Met 
                165                 170                 175     


Gly Ser Met Met Ser Asn Val Ile Ala Val Cys Asp Arg Glu Ala Asp 
            180                 185                 190         


Ile His Ala Tyr Leu Gln Asp Lys Leu Ala His Asn Glu Arg Phe Val 
        195                 200                 205             


Val Arg Ser Lys His Pro Arg Lys Asp Val Glu Ser Gly Leu Tyr Leu 
    210                 215                 220                 


Tyr Asp His Leu Lys Asn Gln Pro Glu Leu Gly Gly Tyr Gln Ile Ser 
225                 230                 235                 240 


Ile Pro Gln Lys Gly Val Val Asp Lys Arg Gly Lys Arg Lys Asn Arg 
                245                 250                 255     


Pro Ala Arg Lys Ala Ser Leu Ser Leu Arg Ser Gly Arg Ile Thr Leu 
            260                 265                 270         


Lys Gln Gly Asn Ile Thr Leu Asn Ala Val Leu Ala Glu Glu Ile Asn 
        275                 280                 285             


Pro Pro Lys Gly Glu Thr Pro Leu Lys Trp Leu Leu Leu Thr Ser Glu 
    290                 295                 300                 


Pro Val Glu Ser Leu Ala Gln Ala Leu Arg Val Ile Asp Ile Tyr Thr 
305                 310                 315                 320 


His Arg Trp Arg Ile Glu Glu Phe His Lys Ala Trp Lys Thr Gly Ala 
                325                 330                 335     


Gly Ala Glu Arg Gln Arg Met Glu Glu Pro Asp Asn Leu Glu Arg Met 
            340                 345                 350         


Val Ser Ile Leu Ser Phe Val Ala Val Arg Leu Leu Gln Leu Arg Glu 
        355                 360                 365             


Ser Phe Thr Pro Pro Gln Ala Leu Arg Ala Gln Gly Leu Leu Lys Glu 
    370                 375                 380                 


Ala Glu His Val Glu Ser Gln Ser Ala Glu Thr Val Leu Thr Pro Asp 
385                 390                 395                 400 


Glu Cys Gln Leu Leu Gly Tyr Leu Asp Lys Gly Lys Arg Lys Arg Lys 
                405                 410                 415     


Glu Lys Ala Gly Ser Leu Gln Trp Ala Tyr Met Ala Ile Ala Arg Leu 
            420                 425                 430         


Gly Gly Phe Met Asp Ser Lys Arg Thr Gly Ile Ala Ser Trp Gly Ala 
        435                 440                 445             


Leu Trp Glu Gly Trp Glu Ala Leu Gln Ser Lys Leu Asp Gly Phe Leu 
    450                 455                 460                 


Ala Ala Lys Asp Leu Met Ala Gln Gly Ile Lys Ile Ser Gly Ser Glu 
465                 470                 475                 480 


Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Met Asp Lys Lys 
                485                 490                 495     


Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val 
            500                 505                 510         


Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly 
        515                 520                 525             


Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu 
    530                 535                 540                 


Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala 
545                 550                 555                 560 


Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu 
                565                 570                 575     


Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe His Arg 
            580                 585                 590         


Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu Arg His 
        595                 600                 605             


Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr 
    610                 615                 620                 


Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys 
625                 630                 635                 640 


Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe 
                645                 650                 655     


Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp 
            660                 665                 670         


Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe 
        675                 680                 685             


Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu 
    690                 695                 700                 


Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln 
705                 710                 715                 720 


Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu 
                725                 730                 735     


Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu 
            740                 745                 750         


Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp 
        755                 760                 765             


Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala 
    770                 775                 780                 


Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val 
785                 790                 795                 800 


Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg 
                805                 810                 815     


Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg 
            820                 825                 830         


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


Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe 
    850                 855                 860                 


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu 
865                 870                 875                 880 


Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr 
                885                 890                 895     


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu His 
            900                 905                 910         


Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn 
        915                 920                 925             


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
    930                 935                 940                 


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys 
945                 950                 955                 960 


Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
                965                 970                 975     


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys 
            980                 985                 990         


Asn Leu Pro Asn Glu Lys Val Leu  Pro Lys His Ser Leu  Leu Tyr Glu 
        995                 1000                 1005             


Tyr Phe  Thr Val Tyr Asn Glu  Leu Thr Lys Val Lys  Tyr Val Thr 
    1010                 1015                 1020             


Glu Gly  Met Arg Lys Pro Ala  Phe Leu Ser Gly Glu  Gln Lys Lys 
    1025                 1030                 1035             


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


Lys Gln  Leu Lys Glu Asp Tyr  Phe Lys Lys Ile Glu  Cys Phe Asp 
    1055                 1060                 1065             


Ser Val  Glu Ile Ser Gly Val  Glu Asp Arg Phe Asn  Ala Ser Leu 
    1070                 1075                 1080             


Gly Thr  Tyr His Asp Leu Leu  Lys Ile Ile Lys Asp  Lys Asp Phe 
    1085                 1090                 1095             


Leu Asp  Asn Glu Glu Asn Glu  Asp Ile Leu Glu Asp  Ile Val Leu 
    1100                 1105                 1110             


Thr Leu  Thr Leu Phe Glu Asp  Arg Glu Met Ile Glu  Glu Arg Leu 
    1115                 1120                 1125             


Lys Thr  Tyr Ala His Leu Phe  Asp Asp Lys Val Met  Lys Gln Leu 
    1130                 1135                 1140             


Lys Arg  Arg Arg Tyr Thr Gly  Trp Gly Arg Leu Ser  Arg Lys Leu 
    1145                 1150                 1155             


Ile Asn  Gly Ile Arg Asp Lys  Gln Ser Gly Lys Thr  Ile Leu Asp 
    1160                 1165                 1170             


Phe Leu  Lys Ser Asp Gly Phe  Ala Asn Arg Asn Phe  Met Gln Leu 
    1175                 1180                 1185             


Ile His  Asp Asp Ser Leu Thr  Phe Lys Glu Asp Ile  Gln Lys Ala 
    1190                 1195                 1200             


Gln Val  Ser Gly Gln Gly Asp  Ser Leu His Glu His  Ile Ala Asn 
    1205                 1210                 1215             


Leu Ala  Gly Ser Pro Ala Ile  Lys Lys Gly Ile Leu  Gln Thr Val 
    1220                 1225                 1230             


Lys Val  Val Asp Glu Leu Val  Lys Val Met Gly Arg  His Lys Pro 
    1235                 1240                 1245             


Glu Asn  Ile Val Ile Glu Met  Ala Arg Glu Asn Gln  Thr Thr Gln 
    1250                 1255                 1260             


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


Gly Ile  Lys Glu Leu Gly Ser  Gln Ile Leu Lys Glu  His Pro Val 
    1280                 1285                 1290             


Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu Tyr Leu  Tyr Tyr Leu 
    1295                 1300                 1305             


Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln Glu Leu  Asp Ile Asn 
    1310                 1315                 1320             


Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile Val Pro  Gln Ser Phe 
    1325                 1330                 1335             


Leu Lys  Asp Asp Ser Ile Asp  Asn Lys Val Leu Thr  Arg Ser Asp 
    1340                 1345                 1350             


Lys Asn  Arg Gly Lys Ser Asp  Asn Val Pro Ser Glu  Glu Val Val 
    1355                 1360                 1365             


Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu Leu Asn  Ala Lys Leu 
    1370                 1375                 1380             


Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr Lys Ala  Glu Arg Gly 
    1385                 1390                 1395             


Gly Leu  Ser Glu Leu Asp Lys  Ala Gly Phe Ile Lys  Arg Gln Leu 
    1400                 1405                 1410             


Val Glu  Thr Arg Gln Ile Thr  Lys His Val Ala Gln  Ile Leu Asp 
    1415                 1420                 1425             


Ser Arg  Met Asn Thr Lys Tyr  Asp Glu Asn Asp Lys  Leu Ile Arg 
    1430                 1435                 1440             


Glu Val  Lys Val Ile Thr Leu  Lys Ser Lys Leu Val  Ser Asp Phe 
    1445                 1450                 1455             


Arg Lys  Asp Phe Gln Phe Tyr  Lys Val Arg Glu Ile  Asn Asn Tyr 
    1460                 1465                 1470             


His His  Ala His Asp Ala Tyr  Leu Asn Ala Val Val  Gly Thr Ala 
    1475                 1480                 1485             


Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser Glu Phe  Val Tyr Gly 
    1490                 1495                 1500             


Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met Ile Ala  Lys Ser Glu 
    1505                 1510                 1515             


Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Tyr Phe Phe  Tyr Ser Asn 
    1520                 1525                 1530             


Ile Met  Asn Phe Phe Lys Thr  Glu Ile Thr Leu Ala  Asn Gly Glu 
    1535                 1540                 1545             


Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn Gly Glu  Thr Gly Glu 
    1550                 1555                 1560             


Ile Val  Trp Asp Lys Gly Arg  Asp Phe Ala Thr Val  Arg Lys Val 
    1565                 1570                 1575             


Leu Ser  Met Pro Gln Val Asn  Ile Val Lys Lys Thr  Glu Val Gln 
    1580                 1585                 1590             


Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu Pro Lys  Arg Asn Ser 
    1595                 1600                 1605             


Asp Lys  Leu Ile Ala Arg Lys  Lys Asp Trp Asp Pro  Lys Lys Tyr 
    1610                 1615                 1620             


Gly Gly  Phe Asp Ser Pro Thr  Val Ala Tyr Ser Val  Leu Val Val 
    1625                 1630                 1635             


Ala Lys  Val Glu Lys Gly Lys  Ser Lys Lys Leu Lys  Ser Val Lys 
    1640                 1645                 1650             


Glu Leu  Leu Gly Ile Thr Ile  Met Glu Arg Ser Ser  Phe Glu Lys 
    1655                 1660                 1665             


Asn Pro  Ile Asp Phe Leu Glu  Ala Lys Gly Tyr Lys  Glu Val Lys 
    1670                 1675                 1680             


Lys Asp  Leu Ile Ile Lys Leu  Pro Lys Tyr Ser Leu  Phe Glu Leu 
    1685                 1690                 1695             


Glu Asn  Gly Arg Lys Arg Met  Leu Ala Ser Ala Gly  Glu Leu Gln 
    1700                 1705                 1710             


Lys Gly  Asn Glu Leu Ala Leu  Pro Ser Lys Tyr Val  Asn Phe Leu 
    1715                 1720                 1725             


Tyr Leu  Ala Ser His Tyr Glu  Lys Leu Lys Gly Ser  Pro Glu Asp 
    1730                 1735                 1740             


Asn Glu  Gln Lys Gln Leu Phe  Val Glu Gln His Lys  His Tyr Leu 
    1745                 1750                 1755             


Asp Glu  Ile Ile Glu Gln Ile  Ser Glu Phe Ser Lys  Arg Val Ile 
    1760                 1765                 1770             


Leu Ala  Asp Ala Asn Leu Asp  Lys Val Leu Ser Ala  Tyr Asn Lys 
    1775                 1780                 1785             


His Arg  Asp Lys Pro Ile Arg  Glu Gln Ala Glu Asn  Ile Ile His 
    1790                 1795                 1800             


Leu Phe  Thr Leu Thr Asn Leu  Gly Ala Pro Ala Ala  Phe Lys Tyr 
    1805                 1810                 1815             


Phe Asp  Thr Thr Ile Asp Arg  Lys Arg Tyr Thr Ser  Thr Lys Glu 
    1820                 1825                 1830             


Val Leu  Asp Ala Thr Leu Ile  His Gln Ser Ile Thr  Gly Leu Tyr 
    1835                 1840                 1845             


Glu Thr  Arg Ile Asp Leu Ser  Gln Leu Gly Gly Asp  
    1850                 1855                 1860 


<210>  6
<211>  1368
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: dCas9 (D10A, H840A)

<400>  6

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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


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


<210>  7
<211>  1775
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Himar1C9-dCas9 fusion protein with N-terminus 3xFLAG 
       and SV40 mammalian NLS

<400>  7

Met Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp 
1               5                   10                  15      


Tyr Lys Asp Asp Asp Asp Lys Met Ala Pro Lys Lys Lys Arg Lys Val 
            20                  25                  30          


Gly Ile His Arg Gly Val Pro Gly Gly Ser Gly Ser Met Glu Lys Lys 
        35                  40                  45              


Glu Phe Arg Val Leu Ile Lys Tyr Cys Phe Leu Lys Gly Lys Asn Thr 
    50                  55                  60                  


Val Glu Ala Lys Thr Trp Leu Asp Asn Glu Phe Pro Asp Ser Ala Pro 
65                  70                  75                  80  


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


Met Ser Thr Glu Asp Gly Glu Arg Ser Gly Arg Pro Lys Glu Val Val 
            100                 105                 110         


Thr Asp Glu Asn Ile Lys Lys Ile His Lys Met Ile Leu Asn Asp Arg 
        115                 120                 125             


Lys Met Lys Leu Ile Glu Ile Ala Glu Ala Leu Lys Ile Ser Lys Glu 
    130                 135                 140                 


Arg Val Gly His Ile Ile His Gln Tyr Leu Asp Met Arg Lys Leu Cys 
145                 150                 155                 160 


Ala Lys Trp Val Pro Arg Glu Leu Thr Phe Asp Gln Lys Gln Arg Arg 
                165                 170                 175     


Val Asp Asp Ser Lys Arg Cys Leu Gln Leu Leu Thr Arg Asn Thr Pro 
            180                 185                 190         


Glu Phe Phe Arg Arg Tyr Val Thr Met Asp Glu Thr Trp Leu His His 
        195                 200                 205             


Tyr Thr Pro Glu Ser Asn Arg Gln Ser Ala Glu Trp Thr Ala Thr Gly 
    210                 215                 220                 


Glu Pro Ser Pro Lys Arg Gly Lys Thr Gln Lys Ser Ala Gly Lys Val 
225                 230                 235                 240 


Met Ala Ser Val Phe Trp Asp Ala His Gly Ile Ile Phe Ile Asp Tyr 
                245                 250                 255     


Leu Glu Lys Gly Lys Thr Ile Asn Ser Asp Tyr Tyr Met Ala Leu Leu 
            260                 265                 270         


Glu Arg Leu Lys Val Glu Ile Ala Ala Lys Arg Pro His Met Lys Lys 
        275                 280                 285             


Lys Lys Val Leu Phe His Gln Asp Asn Ala Pro Cys His Lys Ser Leu 
    290                 295                 300                 


Arg Thr Met Ala Lys Ile His Glu Leu Gly Phe Glu Leu Leu Pro His 
305                 310                 315                 320 


Pro Pro Tyr Ser Pro Asp Leu Ala Pro Ser Asp Phe Phe Leu Phe Ser 
                325                 330                 335     


Asp Leu Lys Arg Met Leu Ala Gly Lys Lys Phe Gly Cys Asn Glu Glu 
            340                 345                 350         


Val Ile Ala Glu Thr Glu Ala Tyr Phe Glu Ala Lys Pro Lys Glu Tyr 
        355                 360                 365             


Tyr Gln Asn Gly Ile Lys Lys Leu Glu Gly Arg Tyr Asn Arg Cys Ile 
    370                 375                 380                 


Ala Leu Glu Gly Asn Tyr Val Glu Ser Gly Ser Glu Thr Pro Gly Thr 
385                 390                 395                 400 


Ser Glu Ser Ala Thr Pro Glu Ser Asp Lys Lys Tyr Ser Ile Gly Leu 
                405                 410                 415     


Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr 
            420                 425                 430         


Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His 
        435                 440                 445             


Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu 
    450                 455                 460                 


Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr 
465                 470                 475                 480 


Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu 
                485                 490                 495     


Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe 
            500                 505                 510         


Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn 
        515                 520                 525             


Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His 
    530                 535                 540                 


Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu 
545                 550                 555                 560 


Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu 
                565                 570                 575     


Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe 
            580                 585                 590         


Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile 
        595                 600                 605             


Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser 
    610                 615                 620                 


Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys 
625                 630                 635                 640 


Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr 
                645                 650                 655     


Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln 
            660                 665                 670         


Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln 
        675                 680                 685             


Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser 
    690                 695                 700                 


Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr 
705                 710                 715                 720 


Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His 
                725                 730                 735     


Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu 
            740                 745                 750         


Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly 
        755                 760                 765             


Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys 
    770                 775                 780                 


Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu 
785                 790                 795                 800 


Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser 
                805                 810                 815     


Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg 
            820                 825                 830         


Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu 
        835                 840                 845             


Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg 
    850                 855                 860                 


Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile 
865                 870                 875                 880 


Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln 
                885                 890                 895     


Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu 
            900                 905                 910         


Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr 
        915                 920                 925             


Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro 
    930                 935                 940                 


Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe 
945                 950                 955                 960 


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


Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp 
            980                 985                 990         


Arg Phe Asn Ala Ser Leu Gly Thr  Tyr His Asp Leu Leu  Lys Ile Ile 
        995                 1000                 1005             


Lys Asp  Lys Asp Phe Leu Asp  Asn Glu Glu Asn Glu  Asp Ile Leu 
    1010                 1015                 1020             


Glu Asp  Ile Val Leu Thr Leu  Thr Leu Phe Glu Asp  Arg Glu Met 
    1025                 1030                 1035             


Ile Glu  Glu Arg Leu Lys Thr  Tyr Ala His Leu Phe  Asp Asp Lys 
    1040                 1045                 1050             


Val Met  Lys Gln Leu Lys Arg  Arg Arg Tyr Thr Gly  Trp Gly Arg 
    1055                 1060                 1065             


Leu Ser  Arg Lys Leu Ile Asn  Gly Ile Arg Asp Lys  Gln Ser Gly 
    1070                 1075                 1080             


Lys Thr  Ile Leu Asp Phe Leu  Lys Ser Asp Gly Phe  Ala Asn Arg 
    1085                 1090                 1095             


Asn Phe  Met Gln Leu Ile His  Asp Asp Ser Leu Thr  Phe Lys Glu 
    1100                 1105                 1110             


Asp Ile  Gln Lys Ala Gln Val  Ser Gly Gln Gly Asp  Ser Leu His 
    1115                 1120                 1125             


Glu His  Ile Ala Asn Leu Ala  Gly Ser Pro Ala Ile  Lys Lys Gly 
    1130                 1135                 1140             


Ile Leu  Gln Thr Val Lys Val  Val Asp Glu Leu Val  Lys Val Met 
    1145                 1150                 1155             


Gly Arg  His Lys Pro Glu Asn  Ile Val Ile Glu Met  Ala Arg Glu 
    1160                 1165                 1170             


Asn Gln  Thr Thr Gln Lys Gly  Gln Lys Asn Ser Arg  Glu Arg Met 
    1175                 1180                 1185             


Lys Arg  Ile Glu Glu Gly Ile  Lys Glu Leu Gly Ser  Gln Ile Leu 
    1190                 1195                 1200             


Lys Glu  His Pro Val Glu Asn  Thr Gln Leu Gln Asn  Glu Lys Leu 
    1205                 1210                 1215             


Tyr Leu  Tyr Tyr Leu Gln Asn  Gly Arg Asp Met Tyr  Val Asp Gln 
    1220                 1225                 1230             


Glu Leu  Asp Ile Asn Arg Leu  Ser Asp Tyr Asp Val  Asp Ala Ile 
    1235                 1240                 1245             


Val Pro  Gln Ser Phe Leu Lys  Asp Asp Ser Ile Asp  Asn Lys Val 
    1250                 1255                 1260             


Leu Thr  Arg Ser Asp Lys Asn  Arg Gly Lys Ser Asp  Asn Val Pro 
    1265                 1270                 1275             


Ser Glu  Glu Val Val Lys Lys  Met Lys Asn Tyr Trp  Arg Gln Leu 
    1280                 1285                 1290             


Leu Asn  Ala Lys Leu Ile Thr  Gln Arg Lys Phe Asp  Asn Leu Thr 
    1295                 1300                 1305             


Lys Ala  Glu Arg Gly Gly Leu  Ser Glu Leu Asp Lys  Ala Gly Phe 
    1310                 1315                 1320             


Ile Lys  Arg Gln Leu Val Glu  Thr Arg Gln Ile Thr  Lys His Val 
    1325                 1330                 1335             


Ala Gln  Ile Leu Asp Ser Arg  Met Asn Thr Lys Tyr  Asp Glu Asn 
    1340                 1345                 1350             


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


Leu Val  Ser Asp Phe Arg Lys  Asp Phe Gln Phe Tyr  Lys Val Arg 
    1370                 1375                 1380             


Glu Ile  Asn Asn Tyr His His  Ala His Asp Ala Tyr  Leu Asn Ala 
    1385                 1390                 1395             


Val Val  Gly Thr Ala Leu Ile  Lys Lys Tyr Pro Lys  Leu Glu Ser 
    1400                 1405                 1410             


Glu Phe  Val Tyr Gly Asp Tyr  Lys Val Tyr Asp Val  Arg Lys Met 
    1415                 1420                 1425             


Ile Ala  Lys Ser Glu Gln Glu  Ile Gly Lys Ala Thr  Ala Lys Tyr 
    1430                 1435                 1440             


Phe Phe  Tyr Ser Asn Ile Met  Asn Phe Phe Lys Thr  Glu Ile Thr 
    1445                 1450                 1455             


Leu Ala  Asn Gly Glu Ile Arg  Lys Arg Pro Leu Ile  Glu Thr Asn 
    1460                 1465                 1470             


Gly Glu  Thr Gly Glu Ile Val  Trp Asp Lys Gly Arg  Asp Phe Ala 
    1475                 1480                 1485             


Thr Val  Arg Lys Val Leu Ser  Met Pro Gln Val Asn  Ile Val Lys 
    1490                 1495                 1500             


Lys Thr  Glu Val Gln Thr Gly  Gly Phe Ser Lys Glu  Ser Ile Leu 
    1505                 1510                 1515             


Pro Lys  Arg Asn Ser Asp Lys  Leu Ile Ala Arg Lys  Lys Asp Trp 
    1520                 1525                 1530             


Asp Pro  Lys Lys Tyr Gly Gly  Phe Asp Ser Pro Thr  Val Ala Tyr 
    1535                 1540                 1545             


Ser Val  Leu Val Val Ala Lys  Val Glu Lys Gly Lys  Ser Lys Lys 
    1550                 1555                 1560             


Leu Lys  Ser Val Lys Glu Leu  Leu Gly Ile Thr Ile  Met Glu Arg 
    1565                 1570                 1575             


Ser Ser  Phe Glu Lys Asn Pro  Ile Asp Phe Leu Glu  Ala Lys Gly 
    1580                 1585                 1590             


Tyr Lys  Glu Val Lys Lys Asp  Leu Ile Ile Lys Leu  Pro Lys Tyr 
    1595                 1600                 1605             


Ser Leu  Phe Glu Leu Glu Asn  Gly Arg Lys Arg Met  Leu Ala Ser 
    1610                 1615                 1620             


Ala Gly  Glu Leu Gln Lys Gly  Asn Glu Leu Ala Leu  Pro Ser Lys 
    1625                 1630                 1635             


Tyr Val  Asn Phe Leu Tyr Leu  Ala Ser His Tyr Glu  Lys Leu Lys 
    1640                 1645                 1650             


Gly Ser  Pro Glu Asp Asn Glu  Gln Lys Gln Leu Phe  Val Glu Gln 
    1655                 1660                 1665             


His Lys  His Tyr Leu Asp Glu  Ile Ile Glu Gln Ile  Ser Glu Phe 
    1670                 1675                 1680             


Ser Lys  Arg Val Ile Leu Ala  Asp Ala Asn Leu Asp  Lys Val Leu 
    1685                 1690                 1695             


Ser Ala  Tyr Asn Lys His Arg  Asp Lys Pro Ile Arg  Glu Gln Ala 
    1700                 1705                 1710             


Glu Asn  Ile Ile His Leu Phe  Thr Leu Thr Asn Leu  Gly Ala Pro 
    1715                 1720                 1725             


Ala Ala  Phe Lys Tyr Phe Asp  Thr Thr Ile Asp Arg  Lys Arg Tyr 
    1730                 1735                 1740             


Thr Ser  Thr Lys Glu Val Leu  Asp Ala Thr Leu Ile  His Gln Ser 
    1745                 1750                 1755             


Ile Thr  Gly Leu Tyr Glu Thr  Arg Ile Asp Leu Ser  Gln Leu Gly 
    1760                 1765                 1770             


Gly Asp  
    1775 


<210>  8
<211>  1745
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: Himar1C9-dCas9 fusion protein with C-terminal E. coli 
       SsrA degradation tag

<400>  8

Met Glu Lys Lys Glu Phe Arg Val Leu Ile Lys Tyr Cys Phe Leu Lys 
1               5                   10                  15      


Gly Lys Asn Thr Val Glu Ala Lys Thr Trp Leu Asp Asn Glu Phe Pro 
            20                  25                  30          


Asp Ser Ala Pro Gly Lys Ser Thr Ile Ile Asp Trp Tyr Ala Lys Phe 
        35                  40                  45              


Lys Arg Gly Glu Met Ser Thr Glu Asp Gly Glu Arg Ser Gly Arg Pro 
    50                  55                  60                  


Lys Glu Val Val Thr Asp Glu Asn Ile Lys Lys Ile His Lys Met Ile 
65                  70                  75                  80  


Leu Asn Asp Arg Lys Met Lys Leu Ile Glu Ile Ala Glu Ala Leu Lys 
                85                  90                  95      


Ile Ser Lys Glu Arg Val Gly His Ile Ile His Gln Tyr Leu Asp Met 
            100                 105                 110         


Arg Lys Leu Cys Ala Lys Trp Val Pro Arg Glu Leu Thr Phe Asp Gln 
        115                 120                 125             


Lys Gln Arg Arg Val Asp Asp Ser Lys Arg Cys Leu Gln Leu Leu Thr 
    130                 135                 140                 


Arg Asn Thr Pro Glu Phe Phe Arg Arg Tyr Val Thr Met Asp Glu Thr 
145                 150                 155                 160 


Trp Leu His His Tyr Thr Pro Glu Ser Asn Arg Gln Ser Ala Glu Trp 
                165                 170                 175     


Thr Ala Thr Gly Glu Pro Ser Pro Lys Arg Gly Lys Thr Gln Lys Ser 
            180                 185                 190         


Ala Gly Lys Val Met Ala Ser Val Phe Trp Asp Ala His Gly Ile Ile 
        195                 200                 205             


Phe Ile Asp Tyr Leu Glu Lys Gly Lys Thr Ile Asn Ser Asp Tyr Tyr 
    210                 215                 220                 


Met Ala Leu Leu Glu Arg Leu Lys Val Glu Ile Ala Ala Lys Arg Pro 
225                 230                 235                 240 


His Met Lys Lys Lys Lys Val Leu Phe His Gln Asp Asn Ala Pro Cys 
                245                 250                 255     


His Lys Ser Leu Arg Thr Met Ala Lys Ile His Glu Leu Gly Phe Glu 
            260                 265                 270         


Leu Leu Pro His Pro Pro Tyr Ser Pro Asp Leu Ala Pro Ser Asp Phe 
        275                 280                 285             


Phe Leu Phe Ser Asp Leu Lys Arg Met Leu Ala Gly Lys Lys Phe Gly 
    290                 295                 300                 


Cys Asn Glu Glu Val Ile Ala Glu Thr Glu Ala Tyr Phe Glu Ala Lys 
305                 310                 315                 320 


Pro Lys Glu Tyr Tyr Gln Asn Gly Ile Lys Lys Leu Glu Gly Arg Tyr 
                325                 330                 335     


Asn Arg Cys Ile Ala Leu Glu Gly Asn Tyr Val Glu Ser Gly Ser Glu 
            340                 345                 350         


Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Met Asp Lys Lys 
        355                 360                 365             


Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val 
    370                 375                 380                 


Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly 
385                 390                 395                 400 


Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu 
                405                 410                 415     


Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala 
            420                 425                 430         


Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu 
        435                 440                 445             


Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe His Arg 
    450                 455                 460                 


Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu Arg His 
465                 470                 475                 480 


Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr 
                485                 490                 495     


Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys 
            500                 505                 510         


Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe 
        515                 520                 525             


Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp 
    530                 535                 540                 


Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe 
545                 550                 555                 560 


Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu 
                565                 570                 575     


Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln 
            580                 585                 590         


Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu 
        595                 600                 605             


Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu 
    610                 615                 620                 


Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp 
625                 630                 635                 640 


Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala 
                645                 650                 655     


Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val 
            660                 665                 670         


Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg 
        675                 680                 685             


Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg 
    690                 695                 700                 


Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys 
705                 710                 715                 720 


Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe 
                725                 730                 735     


Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu 
            740                 745                 750         


Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr 
        755                 760                 765             


Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu His 
    770                 775                 780                 


Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn 
785                 790                 795                 800 


Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val 
                805                 810                 815     


Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys 
            820                 825                 830         


Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys 
        835                 840                 845             


Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys 
    850                 855                 860                 


Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu 
865                 870                 875                 880 


Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu 
                885                 890                 895     


Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile 
            900                 905                 910         


Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu 
        915                 920                 925             


Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile 
    930                 935                 940                 


Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp 
945                 950                 955                 960 


Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn 
                965                 970                 975     


Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp 
            980                 985                 990         


Arg Glu Met Ile Glu Glu Arg Leu  Lys Thr Tyr Ala His  Leu Phe Asp 
        995                 1000                 1005             


Asp Lys  Val Met Lys Gln Leu  Lys Arg Arg Arg Tyr  Thr Gly Trp 
    1010                 1015                 1020             


Gly Arg  Leu Ser Arg Lys Leu  Ile Asn Gly Ile Arg  Asp Lys Gln 
    1025                 1030                 1035             


Ser Gly  Lys Thr Ile Leu Asp  Phe Leu Lys Ser Asp  Gly Phe Ala 
    1040                 1045                 1050             


Asn Arg  Asn Phe Met Gln Leu  Ile His Asp Asp Ser  Leu Thr Phe 
    1055                 1060                 1065             


Lys Glu  Asp Ile Gln Lys Ala  Gln Val Ser Gly Gln  Gly Asp Ser 
    1070                 1075                 1080             


Leu His  Glu His Ile Ala Asn  Leu Ala Gly Ser Pro  Ala Ile Lys 
    1085                 1090                 1095             


Lys Gly  Ile Leu Gln Thr Val  Lys Val Val Asp Glu  Leu Val Lys 
    1100                 1105                 1110             


Val Met  Gly Arg His Lys Pro  Glu Asn Ile Val Ile  Glu Met Ala 
    1115                 1120                 1125             


Arg Glu  Asn Gln Thr Thr Gln  Lys Gly Gln Lys Asn  Ser Arg Glu 
    1130                 1135                 1140             


Arg Met  Lys Arg Ile Glu Glu  Gly Ile Lys Glu Leu  Gly Ser Gln 
    1145                 1150                 1155             


Ile Leu  Lys Glu His Pro Val  Glu Asn Thr Gln Leu  Gln Asn Glu 
    1160                 1165                 1170             


Lys Leu  Tyr Leu Tyr Tyr Leu  Gln Asn Gly Arg Asp  Met Tyr Val 
    1175                 1180                 1185             


Asp Gln  Glu Leu Asp Ile Asn  Arg Leu Ser Asp Tyr  Asp Val Asp 
    1190                 1195                 1200             


Ala Ile  Val Pro Gln Ser Phe  Leu Lys Asp Asp Ser  Ile Asp Asn 
    1205                 1210                 1215             


Lys Val  Leu Thr Arg Ser Asp  Lys Asn Arg Gly Lys  Ser Asp Asn 
    1220                 1225                 1230             


Val Pro  Ser Glu Glu Val Val  Lys Lys Met Lys Asn  Tyr Trp Arg 
    1235                 1240                 1245             


Gln Leu  Leu Asn Ala Lys Leu  Ile Thr Gln Arg Lys  Phe Asp Asn 
    1250                 1255                 1260             


Leu Thr  Lys Ala Glu Arg Gly  Gly Leu Ser Glu Leu  Asp Lys Ala 
    1265                 1270                 1275             


Gly Phe  Ile Lys Arg Gln Leu  Val Glu Thr Arg Gln  Ile Thr Lys 
    1280                 1285                 1290             


His Val  Ala Gln Ile Leu Asp  Ser Arg Met Asn Thr  Lys Tyr Asp 
    1295                 1300                 1305             


Glu Asn  Asp Lys Leu Ile Arg  Glu Val Lys Val Ile  Thr Leu Lys 
    1310                 1315                 1320             


Ser Lys  Leu Val Ser Asp Phe  Arg Lys Asp Phe Gln  Phe Tyr Lys 
    1325                 1330                 1335             


Val Arg  Glu Ile Asn Asn Tyr  His His Ala His Asp  Ala Tyr Leu 
    1340                 1345                 1350             


Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys Lys Tyr  Pro Lys Leu 
    1355                 1360                 1365             


Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys Val Tyr  Asp Val Arg 
    1370                 1375                 1380             


Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile Gly Lys  Ala Thr Ala 
    1385                 1390                 1395             


Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn Phe Phe  Lys Thr Glu 
    1400                 1405                 1410             


Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys Arg Pro  Leu Ile Glu 
    1415                 1420                 1425             


Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp Asp Lys  Gly Arg Asp 
    1430                 1435                 1440             


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


Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly Phe Ser  Lys Glu Ser 
    1460                 1465                 1470             


Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu Ile Ala  Arg Lys Lys 
    1475                 1480                 1485             


Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe Asp Ser  Pro Thr Val 
    1490                 1495                 1500             


Ala Tyr  Ser Val Leu Val Val  Ala Lys Val Glu Lys  Gly Lys Ser 
    1505                 1510                 1515             


Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu Gly Ile  Thr Ile Met 
    1520                 1525                 1530             


Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile Asp Phe  Leu Glu Ala 
    1535                 1540                 1545             


Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu Ile Ile  Lys Leu Pro 
    1550                 1555                 1560             


Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly Arg Lys  Arg Met Leu 
    1565                 1570                 1575             


Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn Glu Leu  Ala Leu Pro 
    1580                 1585                 1590             


Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala Ser His  Tyr Glu Lys 
    1595                 1600                 1605             


Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln Lys Gln  Leu Phe Val 
    1610                 1615                 1620             


Glu Gln  His Lys His Tyr Leu  Asp Glu Ile Ile Glu  Gln Ile Ser 
    1625                 1630                 1635             


Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp Ala Asn  Leu Asp Lys 
    1640                 1645                 1650             


Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp Lys Pro  Ile Arg Glu 
    1655                 1660                 1665             


Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr Leu Thr  Asn Leu Gly 
    1670                 1675                 1680             


Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr Thr Ile  Asp Arg Lys 
    1685                 1690                 1695             


Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp Ala Thr  Leu Ile His 
    1700                 1705                 1710             


Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg Ile Asp  Leu Ser Gln 
    1715                 1720                 1725             


Leu Gly  Gly Asp Arg Pro Ala  Ala Asn Asp Glu Asn  Tyr Ala Leu 
    1730                 1735                 1740             


Ala Ala  
    1745 


<210>  9
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Himar1 Transposon inverted repeat

<400>  9
acaggttgga tgataagtcc ccggtct                                           27


<210>  10
<211>  1375
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Himar1 mini-transposon containing chloramphenicol 
       resistance cassette as payload (from plasmid pHimar6)

<400>  10
acaggttgga tgataagtcc ccggtcttcg tatgccgtct tctgcttggc gcgccctcga       60

gcaattgccg accgaatttt tatgtcgtaa agaggggctt tgcagggggt ggactcagaa      120

agatgagaat agatgactat tgtagttgaa acacatagaa agttgctgat atacagaccg      180

atacgcatat cgggatgaac catgagtacg ttcttttctc aaaaaacata aatattcgaa      240

aagagatgca ataaattaag gagaggttat actctagagt agtagattat tttaggaatt      300

tagatgtttt gtatgaaata gatgcttcgt atggaattaa tgaaattttt agtcaggtaa      360

aaaaggtaat aggagaatat tatggagaaa aaaatcactg gatataccac cgttgatata      420

tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat      480

aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac      540

aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt      600

cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc      660

gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc      720

cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat      780

ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc      840

accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg      900

ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat      960

gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat     1020

gagtggcagg gcggggcgta aaaacaatag gccacatgca actgtctaga atgcgagagt     1080

agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt     1140

ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt     1200

tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca     1260

ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacctgcag     1320

ggcgcgccaa gcagaagacg gcatacgaag accggggact tatcatccaa cctgt          1375


<210>  11
<211>  5199
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: DNA coding sequence for Himar1C9-dCas9 fusion protein 
       with XTEN linker

<400>  11
atggaaaaaa aggaatttcg tgttttgata aaatactgtt ttctgaaggg aaaaaataca       60

gtggaagcaa aaacttggct tgataatgag tttccggact ctgccccagg gaaatcaaca      120

ataattgatt ggtatgcaaa attcaagcgt ggtgaaatga gcacggagga cggtgaacgc      180

agtggacgcc cgaaagaggt ggttaccgac gaaaacatca aaaaaatcca caaaatgatt      240

ttgaatgacc gtaaaatgaa gttgatcgag atagcagagg ccttaaagat atcaaaggaa      300

cgtgttggtc atatcattca tcaatatttg gatatgcgga agctctgtgc gaaatgggtg      360

ccgcgcgagc tcacatttga ccaaaaacaa cgacgtgttg atgattctaa gcggtgtttg      420

cagctgttaa ctcgtaatac acccgagttt ttccgtcgat atgtgacaat ggatgaaaca      480

tggctccatc actacactcc tgagtccaat cgacagtcgg ctgagtggac agcgaccggt      540

gaaccgtctc cgaagcgtgg aaagactcaa aagtccgctg gcaaagtaat ggcctctgtt      600

ttttgggatg cgcatggaat aatttttatc gattatcttg agaagggaaa aaccatcaac      660

agtgactatt atatggcgtt attggagcgt ttgaaggtcg aaatcgcggc aaaacggccc      720

cacatgaaga agaaaaaagt gttgttccac caagacaacg caccgtgcca caagtcattg      780

agaacgatgg caaaaattca tgaattgggc ttcgaattgc ttccccaccc gccgtattct      840

ccagatctgg cccccagcga ctttttcttg ttctcagacc tcaaaaggat gctcgcaggg      900

aaaaaatttg gctgcaatga agaggtgatc gccgaaactg aggcctattt tgaggcaaaa      960

ccgaaggagt actaccaaaa tggtatcaaa aaattggaag gtcgttataa tcgttgtatc     1020

gctcttgaag ggaactatgt tgaaagcggt tccgaaactc ccggtacatc agaaagcgcg     1080

acccccgaaa gcatggataa aaagtattct attggtttag ctatcggcac aaatagcgtc     1140

ggatgggcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa ggttctggga     1200

aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt tgacagtgga     1260

gagacagcgg aagcgactcg tctcaaacgg acagctcgta gaaggtatac acgtcggaag     1320

aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt agatgatagt     1380

ttctttcatc gacttgaaga gtcttttttg gtggaagaag acaagaagca tgaacgtcat     1440

cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc aactatctat     1500

catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt aatctatttg     1560

gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga tttaaatcct     1620

gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa tcaattattt     1680

gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc tgcacgattg     1740

agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc     1800

ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa atcaaatttt     1860

gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga tgatttagat     1920

aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc taagaattta     1980

tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac taaggctccc     2040

ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac tcttttaaaa     2100

gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga tcaatcaaaa     2160

aacggatatg caggttatat tgatggggga gctagccaag aagaatttta taaatttatc     2220

aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact aaatcgtgaa     2280

gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca aattcacttg     2340

ggtgagctgc atgctatttt gagaagacaa gaagactttt atccattttt aaaagacaat     2400

cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt attatgttgg tccattggcg     2460

cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat taccccatgg     2520

aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga acgcatgaca     2580

aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt gctttatgag     2640

tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa     2700

ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt caaaacaaat     2760

cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga atgttttgat     2820

agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac ctaccatgat     2880

ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga agatatctta     2940

gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga ggaaagactt     3000

aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg tcgccgttat     3060

actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa gcaatctggc     3120

aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt tatgcagctg     3180

atccatgatg atagtttgac atttaaagaa gacattcaaa aagcacaagt gtctggacaa     3240

ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat taaaaaaggt     3300

attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg gcataagcca     3360

gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat     3420

tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag tcagattctt     3480

aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct ctattatctc     3540

caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt aagtgattat     3600

gatgtcgatg ccattgttcc acaaagtttc cttaaagacg attcaataga caataaggtc     3660

ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga agaagtagtc     3720

aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac tcaacgtaag     3780

tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa agctggtttt     3840

atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca aattttggat     3900

agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt taaagtgatt     3960

accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta taaagtacgt     4020

gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt tggaactgct     4080

ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta taaagtttat     4140

gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac cgcaaaatat     4200

ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc aaatggagag     4260

attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt ctgggataaa     4320

gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag     4380

aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa aagaaattcg     4440

gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg ttttgatagt     4500

ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag     4560

ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc ctttgaaaaa     4620

aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga cttaatcatt     4680

aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat gctggctagt     4740

gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt gaatttttta     4800

tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga acaaaaacaa     4860

ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat cagtgaattt     4920

tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc atataacaaa     4980

catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt tacgttgacg     5040

aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg taaacgatat     5100

acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac tggtctttat     5160

gaaacacgca ttgatttgag tcagctagga ggtgactaa                            5199


<210>  12
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Tn5 transposon inverted repeat

<400>  12
ctgtctctta tacacatct                                                    19


<210>  13
<211>  1041
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Tn5 mini-transposon containing chloramphenicol 
       resistance cassette as payload

<400>  13
ctgtctctta tacacatctc aaccatcatc gatgaatttt ctcgggtgtt ctcgcatatt       60

ggctcgaatt cctgcagccc ctctagagta gtagattatt ttaggaattt agatgttttg      120

tatgaaatag atgcttcgta tggaattaat gaaattttta gtcaggtaaa aaaggtaata      180

ggagaatatt atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca      240

tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt      300

tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc      360

ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat      420

gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga      480

gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct      540

acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg      600

gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga      660

tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta      720

tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga      780

tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg      840

cggggcgtaa aaacaatagg ccacatgcaa ctgtctagaa tgcgagagta gggaactgcc      900

aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tattgaacgg      960

tagcatcttg acgacgcagc ttgccaacga ctacgcacta gccaacaaga gcttcagggt     1020

tgagatgtgt ataagagaca g                                               1041


<210>  14
<211>  120
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Fig. 8 sequence

<400>  14
catgacttct tcaagtccgc catgccggaa ggctatgtgc aggacgcacg atttccttta       60

aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa      120


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

<220>
<223>  Synthetic: peptide linker


<220>
<221>  misc_feature
<222>  (4)..(30)
<223>  Any "Gly Gly Ser" may or may not be present

<400>  15

Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly 
1               5                   10                  15      


Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 
            20                  25                  30  


<210>  16
<211>  18
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: peptide linker

<400>  16

Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly 
1               5                   10                  15      


Gly Ser 
        


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

<220>
<223>  Synthetic: gRNA_1

<400>  17
gtcgttacca gagtcggcca                                                   20


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

<220>
<223>  Synthetic: gRNA_2

<400>  18
tcagtgcttt gctcgttatc                                                   20


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

<220>
<223>  Synthetic: gRNA_3

<400>  19
cgttcctgca catagccttc                                                   20


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

<220>
<223>  Synthetic: gRNA_4

<400>  20
cggcacgtac aaaacgcgtg                                                   20


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

<220>
<223>  Synthetic: gRNA_5

<400>  21
gtcggcgggg tgcttcacgt                                                   20


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

<220>
<223>  Synthetic: gRNA_7

<400>  22
accagagtcg gccaaggtac                                                   20


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

<220>
<223>  Synthetic: gRNA_8

<400>  23
ctgcacatag ccttccggca                                                   20


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

<220>
<223>  Synthetic: gRNA_9

<400>  24
caatgccttt cagctcaatg                                                   20


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

<220>
<223>  Synthetic: gRNA_10

<400>  25
cagctcaatg cggtttacca                                                   20


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

<220>
<223>  Synthetic: gRNA_11

<400>  26
gtaaaccgca ttgagctgaa                                                   20


<210>  27
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: gRNA_12

<400>  27
caatatcctg ggccataagc                                                   20


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

<220>
<223>  Synthetic: gRNA_13

<400>  28
agaacaggac catcaccgat                                                   20


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

<220>
<223>  Synthetic: gRNA_14

<400>  29
gtgctcagat agtgattgtc                                                   20


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

<220>
<223>  Synthetic: gRNA_15

<400>  30
gaactggatg gtgatgtcaa                                                   20


<210>  31
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: gRNA_16

<400>  31
ccttccccga gggcttcaag                                                   20


<210>  32
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: gRNA_18

<400>  32
acgcgatcac atggttctgc                                                   20


<210>  33
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p433

<400>  33
cgcttacaat ttccattcgc cattc                                             25


<210>  34
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p415

<400>  34
ccctgcaaag cccctcttta cg                                                22


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

<220>
<223>  Synthetic: p828

<400>  35
ctgcgcaacc caagtgctac                                                   20


<210>  36
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p829

<400>  36
cagtccagag aaatcggcat tca                                               23


<210>  37
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p923


<220>
<221>  modified_base
<222>  (1)..(1)
<223>  5' biotin

<400>  37
gccataaact gccaggcatc aa                                                22


<210>  38
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p922

<400>  38
ccttcttgcg catctcacg                                                    19


<210>  39
<211>  34
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Adapter_T


<220>
<221>  modified_base
<222>  (1)..(1)
<223>  5' Phosphate

<400>  39
agatcggaag agcacacgtc tgaactccag tcac                                   34


<210>  40
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: Adapter_B


<220>
<221>  misc_feature
<222>  (29)..(30)
<223>  n is a, c, g, or t

<400>  40
gtctcgtggg ctcgggctct tccgatctnn                                        30


<210>  41
<211>  68
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p790

<400>  41
aatgatacgg cgaccaccga gatctacact agatcgccgc cagaccgggg acttatcatc       60

caacctgt                                                                68


<210>  42
<211>  68
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p791

<400>  42
aatgatacgg cgaccaccga gatctacacc tctctatcgc cagaccgggg acttatcatc       60

caacctgt                                                                68


<210>  43
<211>  68
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p792

<400>  43
aatgatacgg cgaccaccga gatctacact atcctctcgc cagaccgggg acttatcatc       60

caacctgt                                                                68


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

<220>
<223>  Synthetic: p793

<400>  44
aatgatacgg cgaccaccga gatctacaca gagtagacgc cagaccgggg acttatcatc       60

caacctgt                                                                68


<210>  45
<211>  68
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p794

<400>  45
aatgatacgg cgaccaccga gatctacacg taaggagcgc cagaccgggg acttatcatc       60

caacctgt                                                                68


<210>  46
<211>  68
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p795

<400>  46
aatgatacgg cgaccaccga gatctacaca ctgcatacgc cagaccgggg acttatcatc       60

caacctgt                                                                68


<210>  47
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p712

<400>  47
cgccagaccg gggacttatc atccaacctg t                                      31


<210>  48
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p713

<400>  48
cggaagagcc cgagcccacg agac                                              24


<210>  49
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p898

<400>  49
tttgagtgag ctgataccgc tc                                                22


<210>  50
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p899

<400>  50
gagcggtatc agctcactca aa                                                22


<210>  51
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic: p900

<400>  51
tcccttaacg tgagttttcg ttcc                                              24


<210>  52
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic: flexible protein linker XTEN35

<400>  52

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


