                         SEQUENCE LISTING

<110> Council of Scientific and Industrial Research, New Delhi
 
<120>  METHOD AND KIT FOR DETECTION OF POLYNUCLEOTIDE

<130>  -

<160>  80    

<170>  PatentIn version 3.5

<210>  1
<211>  1629
<212>  PRT
<213>  Francisella novicida

<400>  1

Met Asn Phe Lys Ile Leu Pro Ile Ala Ile Asp Leu Gly Val Lys Asn 
1               5                   10                  15      


Thr Gly Val Phe Ser Ala Phe Tyr Gln Lys Gly Thr Ser Leu Glu Arg 
            20                  25                  30          


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


Thr Leu Leu Met Asn Asn Arg Thr Ala Arg Arg His Gln Arg Arg Gly 
    50                  55                  60                  


Ile Asp Arg Lys Gln Leu Val Lys Arg Leu Phe Lys Leu Ile Trp Thr 
65                  70                  75                  80  


Glu Gln Leu Asn Leu Glu Trp Asp Lys Asp Thr Gln Gln Ala Ile Ser 
                85                  90                  95      


Phe Leu Phe Asn Arg Arg Gly Phe Ser Phe Ile Thr Asp Gly Tyr Ser 
            100                 105                 110         


Pro Glu Tyr Leu Asn Ile Val Pro Glu Gln Val Lys Ala Ile Leu Met 
        115                 120                 125             


Asp Ile Phe Asp Asp Tyr Asn Gly Glu Asp Asp Leu Asp Ser Tyr Leu 
    130                 135                 140                 


Lys Leu Ala Thr Glu Gln Glu Ser Lys Ile Ser Glu Ile Tyr Asn Lys 
145                 150                 155                 160 


Leu Met Gln Lys Ile Leu Glu Phe Lys Leu Met Lys Leu Cys Thr Asp 
                165                 170                 175     


Ile Lys Asp Asp Lys Val Ser Thr Lys Thr Leu Lys Glu Ile Thr Ser 
            180                 185                 190         


Tyr Glu Phe Glu Leu Leu Ala Asp Tyr Leu Ala Asn Tyr Ser Glu Ser 
        195                 200                 205             


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


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


Lys Arg His Ala Thr Ile Asn Asp Arg Ile Leu Asp Thr Leu Leu Thr 
                245                 250                 255     


Asp Asp Leu Asp Ile Trp Asn Phe Asn Phe Glu Lys Phe Asp Phe Asp 
            260                 265                 270         


Lys Asn Glu Glu Lys Leu Gln Asn Gln Glu Asp Lys Asp His Ile Gln 
        275                 280                 285             


Ala His Leu His His Phe Val Phe Ala Val Asn Lys Ile Lys Ser Glu 
    290                 295                 300                 


Met Ala Ser Gly Gly Arg His Arg Ser Gln Tyr Phe Gln Glu Ile Thr 
305                 310                 315                 320 


Asn Val Leu Asp Glu Asn Asn His Gln Glu Gly Tyr Leu Lys Asn Phe 
                325                 330                 335     


Cys Glu Asn Leu His Asn Lys Lys Tyr Ser Asn Leu Ser Val Lys Asn 
            340                 345                 350         


Leu Val Asn Leu Ile Gly Asn Leu Ser Asn Leu Glu Leu Lys Pro Leu 
        355                 360                 365             


Arg Lys Tyr Phe Asn Asp Lys Ile His Ala Lys Ala Asp His Trp Asp 
    370                 375                 380                 


Glu Gln Lys Phe Thr Glu Thr Tyr Cys His Trp Ile Leu Gly Glu Trp 
385                 390                 395                 400 


Arg Val Gly Val Lys Asp Gln Asp Lys Lys Asp Gly Ala Lys Tyr Ser 
                405                 410                 415     


Tyr Lys Asp Leu Cys Asn Glu Leu Lys Gln Lys Val Thr Lys Ala Gly 
            420                 425                 430         


Leu Val Asp Phe Leu Leu Glu Leu Asp Pro Cys Arg Thr Ile Pro Pro 
        435                 440                 445             


Tyr Leu Asp Asn Asn Asn Arg Lys Pro Pro Lys Cys Gln Ser Leu Ile 
    450                 455                 460                 


Leu Asn Pro Lys Phe Leu Asp Asn Gln Tyr Pro Asn Trp Gln Gln Tyr 
465                 470                 475                 480 


Leu Gln Glu Leu Lys Lys Leu Gln Ser Ile Gln Asn Tyr Leu Asp Ser 
                485                 490                 495     


Phe Glu Thr Asp Leu Lys Val Leu Lys Ser Ser Lys Asp Gln Pro Tyr 
            500                 505                 510         


Phe Val Glu Tyr Lys Ser Ser Asn Gln Gln Ile Ala Ser Gly Gln Arg 
        515                 520                 525             


Asp Tyr Lys Asp Leu Asp Ala Arg Ile Leu Gln Phe Ile Phe Asp Arg 
    530                 535                 540                 


Val Lys Ala Ser Asp Glu Leu Leu Leu Asn Glu Ile Tyr Phe Gln Ala 
545                 550                 555                 560 


Lys Lys Leu Lys Gln Lys Ala Ser Ser Glu Leu Glu Lys Leu Glu Ser 
                565                 570                 575     


Ser Lys Lys Leu Asp Glu Val Ile Ala Asn Ser Gln Leu Ser Gln Ile 
            580                 585                 590         


Leu Lys Ser Gln His Thr Asn Gly Ile Phe Glu Gln Gly Thr Phe Leu 
        595                 600                 605             


His Leu Val Cys Lys Tyr Tyr Lys Gln Arg Gln Arg Ala Arg Asp Ser 
    610                 615                 620                 


Arg Leu Tyr Ile Met Pro Glu Tyr Arg Tyr Asp Lys Lys Leu His Lys 
625                 630                 635                 640 


Tyr Asn Asn Thr Gly Arg Phe Asp Asp Asp Asn Gln Leu Leu Thr Tyr 
                645                 650                 655     


Cys Asn His Lys Pro Arg Gln Lys Arg Tyr Gln Leu Leu Asn Asp Leu 
            660                 665                 670         


Ala Gly Val Leu Gln Val Ser Pro Asn Phe Leu Lys Asp Lys Ile Gly 
        675                 680                 685             


Ser Asp Asp Asp Leu Phe Ile Ser Lys Trp Leu Val Glu His Ile Arg 
    690                 695                 700                 


Gly Phe Lys Lys Ala Cys Glu Asp Ser Leu Lys Ile Gln Lys Asp Asn 
705                 710                 715                 720 


Arg Gly Leu Leu Asn His Lys Ile Asn Ile Ala Arg Asn Thr Lys Gly 
                725                 730                 735     


Lys Cys Glu Lys Glu Ile Phe Asn Leu Ile Cys Lys Ile Glu Gly Ser 
            740                 745                 750         


Glu Asp Lys Lys Gly Asn Tyr Lys His Gly Leu Ala Tyr Glu Leu Gly 
        755                 760                 765             


Val Leu Leu Phe Gly Glu Pro Asn Glu Ala Ser Lys Pro Glu Phe Asp 
    770                 775                 780                 


Arg Lys Ile Lys Lys Phe Asn Ser Ile Tyr Ser Phe Ala Gln Ile Gln 
785                 790                 795                 800 


Gln Ile Ala Phe Ala Glu Arg Lys Gly Asn Ala Asn Thr Cys Ala Val 
                805                 810                 815     


Cys Ser Ala Asp Asn Ala His Arg Met Gln Gln Ile Lys Ile Thr Glu 
            820                 825                 830         


Pro Val Glu Asp Asn Lys Asp Lys Ile Ile Leu Ser Ala Lys Ala Gln 
        835                 840                 845             


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


Met Ala Thr Ile Leu Ala Lys Asn Ile Val Asp Asp Asn Trp Gln Asn 
865                 870                 875                 880 


Ile Lys Gln Val Leu Ser Ala Lys His Gln Leu His Ile Pro Ile Ile 
                885                 890                 895     


Thr Glu Ser Asn Ala Phe Glu Phe Glu Pro Ala Leu Ala Asp Val Lys 
            900                 905                 910         


Gly Lys Ser Leu Lys Asp Arg Arg Lys Lys Ala Leu Glu Arg Ile Ser 
        915                 920                 925             


Pro Glu Asn Ile Phe Lys Asp Lys Asn Asn Arg Ile Lys Glu Phe Ala 
    930                 935                 940                 


Lys Gly Ile Ser Ala Tyr Ser Gly Ala Asn Leu Thr Asp Gly Asp Phe 
945                 950                 955                 960 


Asp Gly Ala Lys Glu Glu Leu Asp His Ile Ile Pro Arg Ser His Lys 
                965                 970                 975     


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


Gly Asp Asn Lys Asn Lys Gly Asn  Arg Ile Phe Cys Leu  Arg Asp Leu 
        995                 1000                 1005             


Ala Asp  Asn Tyr Lys Leu Lys  Gln Phe Glu Thr Thr  Asp Asp Leu 
    1010                 1015                 1020             


Glu Ile  Glu Lys Lys Ile Ala  Asp Thr Ile Trp Asp  Ala Asn Lys 
    1025                 1030                 1035             


Lys Asp  Phe Lys Phe Gly Asn  Tyr Arg Ser Phe Ile  Asn Leu Thr 
    1040                 1045                 1050             


Pro Gln  Glu Gln Lys Ala Phe  Arg His Ala Leu Phe  Leu Ala Asp 
    1055                 1060                 1065             


Glu Asn  Pro Ile Lys Gln Ala  Val Ile Arg Ala Ile  Asn Asn Arg 
    1070                 1075                 1080             


Asn Arg  Thr Phe Val Asn Gly  Thr Gln Arg Tyr Phe  Ala Glu Val 
    1085                 1090                 1095             


Leu Ala  Asn Asn Ile Tyr Leu  Arg Ala Lys Lys Glu  Asn Leu Asn 
    1100                 1105                 1110             


Thr Asp  Lys Ile Ser Phe Asp  Tyr Phe Gly Ile Pro  Thr Ile Gly 
    1115                 1120                 1125             


Asn Gly  Arg Gly Ile Ala Glu  Ile Arg Gln Leu Tyr  Glu Lys Val 
    1130                 1135                 1140             


Asp Ser  Asp Ile Gln Ala Tyr  Ala Lys Gly Asp Lys  Pro Gln Ala 
    1145                 1150                 1155             


Ser Tyr  Ser His Leu Ile Asp  Ala Met Leu Ala Phe  Cys Ile Ala 
    1160                 1165                 1170             


Ala Asp  Glu His Arg Asn Asp  Gly Ser Ile Gly Leu  Glu Ile Asp 
    1175                 1180                 1185             


Lys Asn  Tyr Ser Leu Tyr Pro  Leu Asp Lys Asn Thr  Gly Glu Val 
    1190                 1195                 1200             


Phe Thr  Lys Asp Ile Phe Ser  Gln Ile Lys Ile Thr  Asp Asn Glu 
    1205                 1210                 1215             


Phe Ser  Asp Lys Lys Leu Val  Arg Lys Lys Ala Ile  Glu Gly Phe 
    1220                 1225                 1230             


Asn Thr  His Arg Gln Met Thr  Arg Asp Gly Ile Tyr  Ala Glu Asn 
    1235                 1240                 1245             


Tyr Leu  Pro Ile Leu Ile His  Lys Glu Leu Asn Glu  Val Arg Lys 
    1250                 1255                 1260             


Gly Tyr  Thr Trp Lys Asn Ser  Glu Glu Ile Lys Ile  Phe Lys Gly 
    1265                 1270                 1275             


Lys Lys  Tyr Asp Ile Gln Gln  Leu Asn Asn Leu Val  Tyr Cys Leu 
    1280                 1285                 1290             


Lys Phe  Val Asp Lys Pro Ile  Ser Ile Asp Ile Gln  Ile Ser Thr 
    1295                 1300                 1305             


Leu Glu  Glu Leu Arg Asn Ile  Leu Thr Thr Asn Asn  Ile Ala Ala 
    1310                 1315                 1320             


Thr Ala  Glu Tyr Tyr Tyr Ile  Asn Leu Lys Thr Gln  Lys Leu His 
    1325                 1330                 1335             


Glu Tyr  Tyr Ile Glu Asn Tyr  Asn Thr Ala Leu Gly  Tyr Lys Lys 
    1340                 1345                 1350             


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


Glu Arg  Val Lys Ile Lys Ser  Ile Asp Asp Val Lys  Gln Val Leu 
    1370                 1375                 1380             


Asp Lys  Asp Ser Asn Phe Ile  Ile Gly Lys Ile Thr  Leu Pro Phe 
    1385                 1390                 1395             


Lys Lys  Glu Trp Gln Arg Leu  Tyr Arg Glu Trp Gln  Asn Thr Thr 
    1400                 1405                 1410             


Ile Lys  Asp Asp Tyr Glu Phe  Leu Lys Ser Phe Phe  Asn Val Lys 
    1415                 1420                 1425             


Ser Ile  Thr Lys Leu His Lys  Lys Val Arg Lys Asp  Phe Ser Leu 
    1430                 1435                 1440             


Pro Ile  Ser Thr Asn Glu Gly  Lys Phe Leu Val Lys  Arg Lys Thr 
    1445                 1450                 1455             


Trp Asp  Asn Asn Phe Ile Tyr  Gln Ile Leu Asn Asp  Ser Asp Ser 
    1460                 1465                 1470             


Arg Ala  Asp Gly Thr Lys Pro  Phe Ile Pro Ala Phe  Asp Ile Ser 
    1475                 1480                 1485             


Lys Asn  Glu Ile Val Glu Ala  Ile Ile Asp Ser Phe  Thr Ser Lys 
    1490                 1495                 1500             


Asn Ile  Phe Trp Leu Pro Lys  Asn Ile Glu Leu Gln  Lys Val Asp 
    1505                 1510                 1515             


Asn Lys  Asn Ile Phe Ala Ile  Asp Thr Ser Lys Trp  Phe Glu Val 
    1520                 1525                 1530             


Glu Thr  Pro Ser Asp Leu Arg  Asp Ile Gly Ile Ala  Thr Ile Gln 
    1535                 1540                 1545             


Tyr Lys  Ile Asp Asn Asn Ser  Arg Pro Lys Val Arg  Val Lys Leu 
    1550                 1555                 1560             


Asp Tyr  Val Ile Asp Asp Asp  Ser Lys Ile Asn Tyr  Phe Met Asn 
    1565                 1570                 1575             


His Ser  Leu Leu Lys Ser Arg  Tyr Pro Asp Lys Val  Leu Glu Ile 
    1580                 1585                 1590             


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


Lys Thr  Ile Lys Glu Met Leu  Gly Met Lys Leu Ala  Gly Ile Tyr 
    1610                 1615                 1620             


Asn Glu  Thr Ser Asn Asn 
    1625                 


<210>  2
<211>  1629
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Protein sequence of recombinant dFnCas9 from 
       Francisella novicida

<400>  2

Met Asn Phe Lys Ile Leu Pro Ile Ala Ile Ala Leu Gly Val Lys Asn 
1               5                   10                  15      


Thr Gly Val Phe Ser Ala Phe Tyr Gln Lys Gly Thr Ser Leu Glu Arg 
            20                  25                  30          


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


Thr Leu Leu Met Asn Asn Arg Thr Ala Arg Arg His Gln Arg Arg Gly 
    50                  55                  60                  


Ile Asp Arg Lys Gln Leu Val Lys Arg Leu Phe Lys Leu Ile Trp Thr 
65                  70                  75                  80  


Glu Gln Leu Asn Leu Glu Trp Asp Lys Asp Thr Gln Gln Ala Ile Ser 
                85                  90                  95      


Phe Leu Phe Asn Arg Arg Gly Phe Ser Phe Ile Thr Asp Gly Tyr Ser 
            100                 105                 110         


Pro Glu Tyr Leu Asn Ile Val Pro Glu Gln Val Lys Ala Ile Leu Met 
        115                 120                 125             


Asp Ile Phe Asp Asp Tyr Asn Gly Glu Asp Asp Leu Asp Ser Tyr Leu 
    130                 135                 140                 


Lys Leu Ala Thr Glu Gln Glu Ser Lys Ile Ser Glu Ile Tyr Asn Lys 
145                 150                 155                 160 


Leu Met Gln Lys Ile Leu Glu Phe Lys Leu Met Lys Leu Cys Thr Asp 
                165                 170                 175     


Ile Lys Asp Asp Lys Val Ser Thr Lys Thr Leu Lys Glu Ile Thr Ser 
            180                 185                 190         


Tyr Glu Phe Glu Leu Leu Ala Asp Tyr Leu Ala Asn Tyr Ser Glu Ser 
        195                 200                 205             


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


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


Lys Arg His Ala Thr Ile Asn Asp Arg Ile Leu Asp Thr Leu Leu Thr 
                245                 250                 255     


Asp Asp Leu Asp Ile Trp Asn Phe Asn Phe Glu Lys Phe Asp Phe Asp 
            260                 265                 270         


Lys Asn Glu Glu Lys Leu Gln Asn Gln Glu Asp Lys Asp His Ile Gln 
        275                 280                 285             


Ala His Leu His His Phe Val Phe Ala Val Asn Lys Ile Lys Ser Glu 
    290                 295                 300                 


Met Ala Ser Gly Gly Arg His Arg Ser Gln Tyr Phe Gln Glu Ile Thr 
305                 310                 315                 320 


Asn Val Leu Asp Glu Asn Asn His Gln Glu Gly Tyr Leu Lys Asn Phe 
                325                 330                 335     


Cys Glu Asn Leu His Asn Lys Lys Tyr Ser Asn Leu Ser Val Lys Asn 
            340                 345                 350         


Leu Val Asn Leu Ile Gly Asn Leu Ser Asn Leu Glu Leu Lys Pro Leu 
        355                 360                 365             


Arg Lys Tyr Phe Asn Asp Lys Ile His Ala Lys Ala Asp His Trp Asp 
    370                 375                 380                 


Glu Gln Lys Phe Thr Glu Thr Tyr Cys His Trp Ile Leu Gly Glu Trp 
385                 390                 395                 400 


Arg Val Gly Val Lys Asp Gln Asp Lys Lys Asp Gly Ala Lys Tyr Ser 
                405                 410                 415     


Tyr Lys Asp Leu Cys Asn Glu Leu Lys Gln Lys Val Thr Lys Ala Gly 
            420                 425                 430         


Leu Val Asp Phe Leu Leu Glu Leu Asp Pro Cys Arg Thr Ile Pro Pro 
        435                 440                 445             


Tyr Leu Asp Asn Asn Asn Arg Lys Pro Pro Lys Cys Gln Ser Leu Ile 
    450                 455                 460                 


Leu Asn Pro Lys Phe Leu Asp Asn Gln Tyr Pro Asn Trp Gln Gln Tyr 
465                 470                 475                 480 


Leu Gln Glu Leu Lys Lys Leu Gln Ser Ile Gln Asn Tyr Leu Asp Ser 
                485                 490                 495     


Phe Glu Thr Asp Leu Lys Val Leu Lys Ser Ser Lys Asp Gln Pro Tyr 
            500                 505                 510         


Phe Val Glu Tyr Lys Ser Ser Asn Gln Gln Ile Ala Ser Gly Gln Arg 
        515                 520                 525             


Asp Tyr Lys Asp Leu Asp Ala Arg Ile Leu Gln Phe Ile Phe Asp Arg 
    530                 535                 540                 


Val Lys Ala Ser Asp Glu Leu Leu Leu Asn Glu Ile Tyr Phe Gln Ala 
545                 550                 555                 560 


Lys Lys Leu Lys Gln Lys Ala Ser Ser Glu Leu Glu Lys Leu Glu Ser 
                565                 570                 575     


Ser Lys Lys Leu Asp Glu Val Ile Ala Asn Ser Gln Leu Ser Gln Ile 
            580                 585                 590         


Leu Lys Ser Gln His Thr Asn Gly Ile Phe Glu Gln Gly Thr Phe Leu 
        595                 600                 605             


His Leu Val Cys Lys Tyr Tyr Lys Gln Arg Gln Arg Ala Arg Asp Ser 
    610                 615                 620                 


Arg Leu Tyr Ile Met Pro Glu Tyr Arg Tyr Asp Lys Lys Leu His Lys 
625                 630                 635                 640 


Tyr Asn Asn Thr Gly Arg Phe Asp Asp Asp Asn Gln Leu Leu Thr Tyr 
                645                 650                 655     


Cys Asn His Lys Pro Arg Gln Lys Arg Tyr Gln Leu Leu Asn Asp Leu 
            660                 665                 670         


Ala Gly Val Leu Gln Val Ser Pro Asn Phe Leu Lys Asp Lys Ile Gly 
        675                 680                 685             


Ser Asp Asp Asp Leu Phe Ile Ser Lys Trp Leu Val Glu His Ile Arg 
    690                 695                 700                 


Gly Phe Lys Lys Ala Cys Glu Asp Ser Leu Lys Ile Gln Lys Asp Asn 
705                 710                 715                 720 


Arg Gly Leu Leu Asn His Lys Ile Asn Ile Ala Arg Asn Thr Lys Gly 
                725                 730                 735     


Lys Cys Glu Lys Glu Ile Phe Asn Leu Ile Cys Lys Ile Glu Gly Ser 
            740                 745                 750         


Glu Asp Lys Lys Gly Asn Tyr Lys His Gly Leu Ala Tyr Glu Leu Gly 
        755                 760                 765             


Val Leu Leu Phe Gly Glu Pro Asn Glu Ala Ser Lys Pro Glu Phe Asp 
    770                 775                 780                 


Arg Lys Ile Lys Lys Phe Asn Ser Ile Tyr Ser Phe Ala Gln Ile Gln 
785                 790                 795                 800 


Gln Ile Ala Phe Ala Glu Arg Lys Gly Asn Ala Asn Thr Cys Ala Val 
                805                 810                 815     


Cys Ser Ala Asp Asn Ala His Arg Met Gln Gln Ile Lys Ile Thr Glu 
            820                 825                 830         


Pro Val Glu Asp Asn Lys Asp Lys Ile Ile Leu Ser Ala Lys Ala Gln 
        835                 840                 845             


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


Met Ala Thr Ile Leu Ala Lys Asn Ile Val Asp Asp Asn Trp Gln Asn 
865                 870                 875                 880 


Ile Lys Gln Val Leu Ser Ala Lys His Gln Leu His Ile Pro Ile Ile 
                885                 890                 895     


Thr Glu Ser Asn Ala Phe Glu Phe Glu Pro Ala Leu Ala Asp Val Lys 
            900                 905                 910         


Gly Lys Ser Leu Lys Asp Arg Arg Lys Lys Ala Leu Glu Arg Ile Ser 
        915                 920                 925             


Pro Glu Asn Ile Phe Lys Asp Lys Asn Asn Arg Ile Lys Glu Phe Ala 
    930                 935                 940                 


Lys Gly Ile Ser Ala Tyr Ser Gly Ala Asn Leu Thr Asp Gly Asp Phe 
945                 950                 955                 960 


Asp Gly Ala Lys Glu Glu Leu Asp Ala Ile Ile Pro Arg Ser His Lys 
                965                 970                 975     


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


Gly Asp Asn Lys Asn Lys Gly Asn  Arg Ile Phe Cys Leu  Arg Asp Leu 
        995                 1000                 1005             


Ala Asp  Asn Tyr Lys Leu Lys  Gln Phe Glu Thr Thr  Asp Asp Leu 
    1010                 1015                 1020             


Glu Ile  Glu Lys Lys Ile Ala  Asp Thr Ile Trp Asp  Ala Asn Lys 
    1025                 1030                 1035             


Lys Asp  Phe Lys Phe Gly Asn  Tyr Arg Ser Phe Ile  Asn Leu Thr 
    1040                 1045                 1050             


Pro Gln  Glu Gln Lys Ala Phe  Arg His Ala Leu Phe  Leu Ala Asp 
    1055                 1060                 1065             


Glu Asn  Pro Ile Lys Gln Ala  Val Ile Arg Ala Ile  Asn Asn Arg 
    1070                 1075                 1080             


Asn Arg  Thr Phe Val Asn Gly  Thr Gln Arg Tyr Phe  Ala Glu Val 
    1085                 1090                 1095             


Leu Ala  Asn Asn Ile Tyr Leu  Arg Ala Lys Lys Glu  Asn Leu Asn 
    1100                 1105                 1110             


Thr Asp  Lys Ile Ser Phe Asp  Tyr Phe Gly Ile Pro  Thr Ile Gly 
    1115                 1120                 1125             


Asn Gly  Arg Gly Ile Ala Glu  Ile Arg Gln Leu Tyr  Glu Lys Val 
    1130                 1135                 1140             


Asp Ser  Asp Ile Gln Ala Tyr  Ala Lys Gly Asp Lys  Pro Gln Ala 
    1145                 1150                 1155             


Ser Tyr  Ser His Leu Ile Asp  Ala Met Leu Ala Phe  Cys Ile Ala 
    1160                 1165                 1170             


Ala Asp  Glu His Arg Asn Asp  Gly Ser Ile Gly Leu  Glu Ile Asp 
    1175                 1180                 1185             


Lys Asn  Tyr Ser Leu Tyr Pro  Leu Asp Lys Asn Thr  Gly Glu Val 
    1190                 1195                 1200             


Phe Thr  Lys Asp Ile Phe Ser  Gln Ile Lys Ile Thr  Asp Asn Glu 
    1205                 1210                 1215             


Phe Ser  Asp Lys Lys Leu Val  Arg Lys Lys Ala Ile  Glu Gly Phe 
    1220                 1225                 1230             


Asn Thr  His Arg Gln Met Thr  Arg Asp Gly Ile Tyr  Ala Glu Asn 
    1235                 1240                 1245             


Tyr Leu  Pro Ile Leu Ile His  Lys Glu Leu Asn Glu  Val Arg Lys 
    1250                 1255                 1260             


Gly Tyr  Thr Trp Lys Asn Ser  Glu Glu Ile Lys Ile  Phe Lys Gly 
    1265                 1270                 1275             


Lys Lys  Tyr Asp Ile Gln Gln  Leu Asn Asn Leu Val  Tyr Cys Leu 
    1280                 1285                 1290             


Lys Phe  Val Asp Lys Pro Ile  Ser Ile Asp Ile Gln  Ile Ser Thr 
    1295                 1300                 1305             


Leu Glu  Glu Leu Arg Asn Ile  Leu Thr Thr Asn Asn  Ile Ala Ala 
    1310                 1315                 1320             


Thr Ala  Glu Tyr Tyr Tyr Ile  Asn Leu Lys Thr Gln  Lys Leu His 
    1325                 1330                 1335             


Glu Tyr  Tyr Ile Glu Asn Tyr  Asn Thr Ala Leu Gly  Tyr Lys Lys 
    1340                 1345                 1350             


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


Glu Arg  Val Lys Ile Lys Ser  Ile Asp Asp Val Lys  Gln Val Leu 
    1370                 1375                 1380             


Asp Lys  Asp Ser Asn Phe Ile  Ile Gly Lys Ile Thr  Leu Pro Phe 
    1385                 1390                 1395             


Lys Lys  Glu Trp Gln Arg Leu  Tyr Arg Glu Trp Gln  Asn Thr Thr 
    1400                 1405                 1410             


Ile Lys  Asp Asp Tyr Glu Phe  Leu Lys Ser Phe Phe  Asn Val Lys 
    1415                 1420                 1425             


Ser Ile  Thr Lys Leu His Lys  Lys Val Arg Lys Asp  Phe Ser Leu 
    1430                 1435                 1440             


Pro Ile  Ser Thr Asn Glu Gly  Lys Phe Leu Val Lys  Arg Lys Thr 
    1445                 1450                 1455             


Trp Asp  Asn Asn Phe Ile Tyr  Gln Ile Leu Asn Asp  Ser Asp Ser 
    1460                 1465                 1470             


Arg Ala  Asp Gly Thr Lys Pro  Phe Ile Pro Ala Phe  Asp Ile Ser 
    1475                 1480                 1485             


Lys Asn  Glu Ile Val Glu Ala  Ile Ile Asp Ser Phe  Thr Ser Lys 
    1490                 1495                 1500             


Asn Ile  Phe Trp Leu Pro Lys  Asn Ile Glu Leu Gln  Lys Val Asp 
    1505                 1510                 1515             


Asn Lys  Asn Ile Phe Ala Ile  Asp Thr Ser Lys Trp  Phe Glu Val 
    1520                 1525                 1530             


Glu Thr  Pro Ser Asp Leu Arg  Asp Ile Gly Ile Ala  Thr Ile Gln 
    1535                 1540                 1545             


Tyr Lys  Ile Asp Asn Asn Ser  Arg Pro Lys Val Arg  Val Lys Leu 
    1550                 1555                 1560             


Asp Tyr  Val Ile Asp Asp Asp  Ser Lys Ile Asn Tyr  Phe Met Asn 
    1565                 1570                 1575             


His Ser  Leu Leu Lys Ser Arg  Tyr Pro Asp Lys Val  Leu Glu Ile 
    1580                 1585                 1590             


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


Lys Thr  Ile Lys Glu Met Leu  Gly Met Lys Leu Ala  Gly Ile Tyr 
    1610                 1615                 1620             


Asn Glu  Thr Ser Asn Asn 
    1625                 


<210>  3
<211>  1899
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Protein sequence of recombinant dFnCas9 from 
       Francisella novicida coupled with Green Fluorescent Protein

<400>  3

Val Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val 
1               5                   10                  15      


Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Arg Gly Glu 
            20                  25                  30          


Gly Glu Gly Asp Ala Thr Asn Gly Lys Leu Thr Leu Lys Phe Ile Cys 
        35                  40                  45              


Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu 
    50                  55                  60                  


Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Gln 
65                  70                  75                  80  


His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg 
                85                  90                  95      


Thr Ile Ser Phe Lys Asp Asp Gly Thr Tyr Lys Thr Arg Ala Glu Val 
            100                 105                 110         


Lys Phe Glu Gly Asp Thr Leu Val Asn Arg Ile Glu Leu Lys Gly Ile 
        115                 120                 125             


Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn 
    130                 135                 140                 


Phe Asn Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly 
145                 150                 155                 160 


Ile Lys Ala Asn Phe Lys Ile Arg His Asn Val Glu Asp Gly Ser Val 
                165                 170                 175     


Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro 
            180                 185                 190         


Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Lys Leu Ser 
        195                 200                 205             


Lys Asp Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val 
    210                 215                 220                 


Thr Ala Ala Gly Ile Thr Leu Gly Met Asp Glu Leu Tyr Lys Gly Ile 
225                 230                 235                 240 


Glu Glu Asn Leu Tyr Phe Gln Ser Asn Ala Met Tyr Pro Tyr Asp Val 
                245                 250                 255     


Pro Asp Tyr Ala Ser Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Asn 
            260                 265                 270         


Phe Lys Ile Leu Pro Ile Ala Ile Ala Leu Gly Val Lys Asn Thr Gly 
        275                 280                 285             


Val Phe Ser Ala Phe Tyr Gln Lys Gly Thr Ser Leu Glu Arg Leu Asp 
    290                 295                 300                 


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


Leu Met Asn Asn Arg Thr Ala Arg Arg His Gln Arg Arg Gly Ile Asp 
                325                 330                 335     


Arg Lys Gln Leu Val Lys Arg Leu Phe Lys Leu Ile Trp Thr Glu Gln 
            340                 345                 350         


Leu Asn Leu Glu Trp Asp Lys Asp Thr Gln Gln Ala Ile Ser Phe Leu 
        355                 360                 365             


Phe Asn Arg Arg Gly Phe Ser Phe Ile Thr Asp Gly Tyr Ser Pro Glu 
    370                 375                 380                 


Tyr Leu Asn Ile Val Pro Glu Gln Val Lys Ala Ile Leu Met Asp Ile 
385                 390                 395                 400 


Phe Asp Asp Tyr Asn Gly Glu Asp Asp Leu Asp Ser Tyr Leu Lys Leu 
                405                 410                 415     


Ala Thr Glu Gln Glu Ser Lys Ile Ser Glu Ile Tyr Asn Lys Leu Met 
            420                 425                 430         


Gln Lys Ile Leu Glu Phe Lys Leu Met Lys Leu Cys Thr Asp Ile Lys 
        435                 440                 445             


Asp Asp Lys Val Ser Thr Lys Thr Leu Lys Glu Ile Thr Ser Tyr Glu 
    450                 455                 460                 


Phe Glu Leu Leu Ala Asp Tyr Leu Ala Asn Tyr Ser Glu Ser Leu Lys 
465                 470                 475                 480 


Thr Gln Lys Phe Ser Tyr Thr Asp Lys Gln Gly Asn Leu Lys Glu Leu 
                485                 490                 495     


Ser Tyr Tyr His His Asp Lys Tyr Asn Ile Gln Glu Phe Leu Lys Arg 
            500                 505                 510         


His Ala Thr Ile Asn Asp Arg Ile Leu Asp Thr Leu Leu Thr Asp Asp 
        515                 520                 525             


Leu Asp Ile Trp Asn Phe Asn Phe Glu Lys Phe Asp Phe Asp Lys Asn 
    530                 535                 540                 


Glu Glu Lys Leu Gln Asn Gln Glu Asp Lys Asp His Ile Gln Ala His 
545                 550                 555                 560 


Leu His His Phe Val Phe Ala Val Asn Lys Ile Lys Ser Glu Met Ala 
                565                 570                 575     


Ser Gly Gly Arg His Arg Ser Gln Tyr Phe Gln Glu Ile Thr Asn Val 
            580                 585                 590         


Leu Asp Glu Asn Asn His Gln Glu Gly Tyr Leu Lys Asn Phe Cys Glu 
        595                 600                 605             


Asn Leu His Asn Lys Lys Tyr Ser Asn Leu Ser Val Lys Asn Leu Val 
    610                 615                 620                 


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


Tyr Phe Asn Asp Lys Ile His Ala Lys Ala Asp His Trp Asp Glu Gln 
                645                 650                 655     


Lys Phe Thr Glu Thr Tyr Cys His Trp Ile Leu Gly Glu Trp Arg Val 
            660                 665                 670         


Gly Val Lys Asp Gln Asp Lys Lys Asp Gly Ala Lys Tyr Ser Tyr Lys 
        675                 680                 685             


Asp Leu Cys Asn Glu Leu Lys Gln Lys Val Thr Lys Ala Gly Leu Val 
    690                 695                 700                 


Asp Phe Leu Leu Glu Leu Asp Pro Cys Arg Thr Ile Pro Pro Tyr Leu 
705                 710                 715                 720 


Asp Asn Asn Asn Arg Lys Pro Pro Lys Cys Gln Ser Leu Ile Leu Asn 
                725                 730                 735     


Pro Lys Phe Leu Asp Asn Gln Tyr Pro Asn Trp Gln Gln Tyr Leu Gln 
            740                 745                 750         


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


Thr Asp Leu Lys Val Leu Lys Ser Ser Lys Asp Gln Pro Tyr Phe Val 
    770                 775                 780                 


Glu Tyr Lys Ser Ser Asn Gln Gln Ile Ala Ser Gly Gln Arg Asp Tyr 
785                 790                 795                 800 


Lys Asp Leu Asp Ala Arg Ile Leu Gln Phe Ile Phe Asp Arg Val Lys 
                805                 810                 815     


Ala Ser Asp Glu Leu Leu Leu Asn Glu Ile Tyr Phe Gln Ala Lys Lys 
            820                 825                 830         


Leu Lys Gln Lys Ala Ser Ser Glu Leu Glu Lys Leu Glu Ser Ser Lys 
        835                 840                 845             


Lys Leu Asp Glu Val Ile Ala Asn Ser Gln Leu Ser Gln Ile Leu Lys 
    850                 855                 860                 


Ser Gln His Thr Asn Gly Ile Phe Glu Gln Gly Thr Phe Leu His Leu 
865                 870                 875                 880 


Val Cys Lys Tyr Tyr Lys Gln Arg Gln Arg Ala Arg Asp Ser Arg Leu 
                885                 890                 895     


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


Asn Thr Gly Arg Phe Asp Asp Asp Asn Gln Leu Leu Thr Tyr Cys Asn 
        915                 920                 925             


His Lys Pro Arg Gln Lys Arg Tyr Gln Leu Leu Asn Asp Leu Ala Gly 
    930                 935                 940                 


Val Leu Gln Val Ser Pro Asn Phe Leu Lys Asp Lys Ile Gly Ser Asp 
945                 950                 955                 960 


Asp Asp Leu Phe Ile Ser Lys Trp Leu Val Glu His Ile Arg Gly Phe 
                965                 970                 975     


Lys Lys Ala Cys Glu Asp Ser Leu Lys Ile Gln Lys Asp Asn Arg Gly 
            980                 985                 990         


Leu Leu Asn His Lys Ile Asn Ile  Ala Arg Asn Thr Lys  Gly Lys Cys 
        995                 1000                 1005             


Glu Lys  Glu Ile Phe Asn Leu  Ile Cys Lys Ile Glu  Gly Ser Glu 
    1010                 1015                 1020             


Asp Lys  Lys Gly Asn Tyr Lys  His Gly Leu Ala Tyr  Glu Leu Gly 
    1025                 1030                 1035             


Val Leu  Leu Phe Gly Glu Pro  Asn Glu Ala Ser Lys  Pro Glu Phe 
    1040                 1045                 1050             


Asp Arg  Lys Ile Lys Lys Phe  Asn Ser Ile Tyr Ser  Phe Ala Gln 
    1055                 1060                 1065             


Ile Gln  Gln Ile Ala Phe Ala  Glu Arg Lys Gly Asn  Ala Asn Thr 
    1070                 1075                 1080             


Cys Ala  Val Cys Ser Ala Asp  Asn Ala His Arg Met  Gln Gln Ile 
    1085                 1090                 1095             


Lys Ile  Thr Glu Pro Val Glu  Asp Asn Lys Asp Lys  Ile Ile Leu 
    1100                 1105                 1110             


Ser Ala  Lys Ala Gln Arg Leu  Pro Ala Ile Pro Thr  Arg Ile Val 
    1115                 1120                 1125             


Asp Gly  Ala Val Lys Lys Met  Ala Thr Ile Leu Ala  Lys Asn Ile 
    1130                 1135                 1140             


Val Asp  Asp Asn Trp Gln Asn  Ile Lys Gln Val Leu  Ser Ala Lys 
    1145                 1150                 1155             


His Gln  Leu His Ile Pro Ile  Ile Thr Glu Ser Asn  Ala Phe Glu 
    1160                 1165                 1170             


Phe Glu  Pro Ala Leu Ala Asp  Val Lys Gly Lys Ser  Leu Lys Asp 
    1175                 1180                 1185             


Arg Arg  Lys Lys Ala Leu Glu  Arg Ile Ser Pro Glu  Asn Ile Phe 
    1190                 1195                 1200             


Lys Asp  Lys Asn Asn Arg Ile  Lys Glu Phe Ala Lys  Gly Ile Ser 
    1205                 1210                 1215             


Ala Tyr  Ser Gly Ala Asn Leu  Thr Asp Gly Asp Phe  Asp Gly Ala 
    1220                 1225                 1230             


Lys Glu  Glu Leu Asp Ala Ile  Ile Pro Arg Ser His  Lys Lys Tyr 
    1235                 1240                 1245             


Gly Thr  Leu Asn Asp Glu Ala  Asn Leu Ile Cys Val  Thr Arg Gly 
    1250                 1255                 1260             


Asp Asn  Lys Asn Lys Gly Asn  Arg Ile Phe Cys Leu  Arg Asp Leu 
    1265                 1270                 1275             


Ala Asp  Asn Tyr Lys Leu Lys  Gln Phe Glu Thr Thr  Asp Asp Leu 
    1280                 1285                 1290             


Glu Ile  Glu Lys Lys Ile Ala  Asp Thr Ile Trp Asp  Ala Asn Lys 
    1295                 1300                 1305             


Lys Asp  Phe Lys Phe Gly Asn  Tyr Arg Ser Phe Ile  Asn Leu Thr 
    1310                 1315                 1320             


Pro Gln  Glu Gln Lys Ala Phe  Arg His Ala Leu Phe  Leu Ala Asp 
    1325                 1330                 1335             


Glu Asn  Pro Ile Lys Gln Ala  Val Ile Arg Ala Ile  Asn Asn Arg 
    1340                 1345                 1350             


Asn Arg  Thr Phe Val Asn Gly  Thr Gln Arg Tyr Phe  Ala Glu Val 
    1355                 1360                 1365             


Leu Ala  Asn Asn Ile Tyr Leu  Arg Ala Lys Lys Glu  Asn Leu Asn 
    1370                 1375                 1380             


Thr Asp  Lys Ile Ser Phe Asp  Tyr Phe Gly Ile Pro  Thr Ile Gly 
    1385                 1390                 1395             


Asn Gly  Arg Gly Ile Ala Glu  Ile Arg Gln Leu Tyr  Glu Lys Val 
    1400                 1405                 1410             


Asp Ser  Asp Ile Gln Ala Tyr  Ala Lys Gly Asp Lys  Pro Gln Ala 
    1415                 1420                 1425             


Ser Tyr  Ser His Leu Ile Asp  Ala Met Leu Ala Phe  Cys Ile Ala 
    1430                 1435                 1440             


Ala Asp  Glu His Arg Asn Asp  Gly Ser Ile Gly Leu  Glu Ile Asp 
    1445                 1450                 1455             


Lys Asn  Tyr Ser Leu Tyr Pro  Leu Asp Lys Asn Thr  Gly Glu Val 
    1460                 1465                 1470             


Phe Thr  Lys Asp Ile Phe Ser  Gln Ile Lys Ile Thr  Asp Asn Glu 
    1475                 1480                 1485             


Phe Ser  Asp Lys Lys Leu Val  Arg Lys Lys Ala Ile  Glu Gly Phe 
    1490                 1495                 1500             


Asn Thr  His Arg Gln Met Thr  Arg Asp Gly Ile Tyr  Ala Glu Asn 
    1505                 1510                 1515             


Tyr Leu  Pro Ile Leu Ile His  Lys Glu Leu Asn Glu  Val Arg Lys 
    1520                 1525                 1530             


Gly Tyr  Thr Trp Lys Asn Ser  Glu Glu Ile Lys Ile  Phe Lys Gly 
    1535                 1540                 1545             


Lys Lys  Tyr Asp Ile Gln Gln  Leu Asn Asn Leu Val  Tyr Cys Leu 
    1550                 1555                 1560             


Lys Phe  Val Asp Lys Pro Ile  Ser Ile Asp Ile Gln  Ile Ser Thr 
    1565                 1570                 1575             


Leu Glu  Glu Leu Arg Asn Ile  Leu Thr Thr Asn Asn  Ile Ala Ala 
    1580                 1585                 1590             


Thr Ala  Glu Tyr Tyr Tyr Ile  Asn Leu Lys Thr Gln  Lys Leu His 
    1595                 1600                 1605             


Glu Tyr  Tyr Ile Glu Asn Tyr  Asn Thr Ala Leu Gly  Tyr Lys Lys 
    1610                 1615                 1620             


Tyr Ser  Lys Glu Met Glu Phe  Leu Arg Ser Leu Ala  Tyr Arg Ser 
    1625                 1630                 1635             


Glu Arg  Val Lys Ile Lys Ser  Ile Asp Asp Val Lys  Gln Val Leu 
    1640                 1645                 1650             


Asp Lys  Asp Ser Asn Phe Ile  Ile Gly Lys Ile Thr  Leu Pro Phe 
    1655                 1660                 1665             


Lys Lys  Glu Trp Gln Arg Leu  Tyr Arg Glu Trp Gln  Asn Thr Thr 
    1670                 1675                 1680             


Ile Lys  Asp Asp Tyr Glu Phe  Leu Lys Ser Phe Phe  Asn Val Lys 
    1685                 1690                 1695             


Ser Ile  Thr Lys Leu His Lys  Lys Val Arg Lys Asp  Phe Ser Leu 
    1700                 1705                 1710             


Pro Ile  Ser Thr Asn Glu Gly  Lys Phe Leu Val Lys  Arg Lys Thr 
    1715                 1720                 1725             


Trp Asp  Asn Asn Phe Ile Tyr  Gln Ile Leu Asn Asp  Ser Asp Ser 
    1730                 1735                 1740             


Arg Ala  Asp Gly Thr Lys Pro  Phe Ile Pro Ala Phe  Asp Ile Ser 
    1745                 1750                 1755             


Lys Asn  Glu Ile Val Glu Ala  Ile Ile Asp Ser Phe  Thr Ser Lys 
    1760                 1765                 1770             


Asn Ile  Phe Trp Leu Pro Lys  Asn Ile Glu Leu Gln  Lys Val Asp 
    1775                 1780                 1785             


Asn Lys  Asn Ile Phe Ala Ile  Asp Thr Ser Lys Trp  Phe Glu Val 
    1790                 1795                 1800             


Glu Thr  Pro Ser Asp Leu Arg  Asp Ile Gly Ile Ala  Thr Ile Gln 
    1805                 1810                 1815             


Tyr Lys  Ile Asp Asn Asn Ser  Arg Pro Lys Val Arg  Val Lys Leu 
    1820                 1825                 1830             


Asp Tyr  Val Ile Asp Asp Asp  Ser Lys Ile Asn Tyr  Phe Met Asn 
    1835                 1840                 1845             


His Ser  Leu Leu Lys Ser Arg  Tyr Pro Asp Lys Val  Leu Glu Ile 
    1850                 1855                 1860             


Leu Lys  Gln Ser Thr Ile Ile  Glu Phe Glu Ser Ser  Gly Phe Asn 
    1865                 1870                 1875             


Lys Thr  Ile Lys Glu Met Leu  Gly Met Lys Leu Ala  Gly Ile Tyr 
    1880                 1885                 1890             


Asn Glu  Thr Ser Asn Asn 
    1895                 


<210>  4
<211>  502
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic DNA containing portion of human gene encoding for 
       Glycogen storage disease type I

<400>  4
agcataggag atggaatttg ttaaataaat gaattatgga taacgaatgg atggtaagat       60

gggtggatgg atggggggtg aacggatgga tggggggtga atggatggat gaatgggtag      120

atgggtggat agggggatgg ctgggtggct gggtagatga tgcactgtct cccagatgag      180

gaccttttca cctttactcc attctctttc ctgcccttta gggagcccct ctggccatgc      240

catgggcaca acaggtgtat actacgtgat ggtcacatct actctttcca tctttcaggg      300

aaagataaag ccgacctaca gatttcggta agaactcacc actggggtgt aggtggtgga      360

gggcaggagg cagctctctc tgtagctgac acaccacgta ttcttcctca catcccccta      420

gcccgctccc acacctgggc agccgctgat taagagttgt ggcactttgg atagggataa      480

acctcagagt cagggaatgt tt                                               502


<210>  5
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic DNA for SgRNA seq.1 against Glycogen storage disease 
       type I

<400>  5
ggccatgcca tgggaacaac                                                   20


<210>  6
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic DNA for SgRNA seq.2 against Glycogen storage disease 
       type I

<400>  6
ggccatgcca tgagcacaac                                                   20


<210>  7
<211>  502
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic DNA containing portion of human gene encoding for 
       Glanzmann thrombosthenia

<400>  7
tcccagggac caagcgtggc ccacagaggc ccacagcaca cccggacccc tctccctcct       60

cccatcccct ctgccccgtg ggtcccgggg aggccgggcc agagaccaga gagcctgctc      120

actacgagaa ctggatcctg aagcctcgag ggctgctcgg gctctggcag gaagatctgt      180

ctgcgatccc gcttgtgatg ggccgggtga atgggggagg ggctggggat gggcagcccc      240

cagtccacct cggggggcaa aggagtggtc aggcccaggt ctcccccgaa ccccagccca      300

cagaggtgcc ccggtggttg gtctggggcc gccttcccag gtctttcttc cacccagctc      360

ttaccttgag agggttgaca ggaggctgtg ggaagcactg aaggcccccc tggggctgta      420

tatccaggat gtagagcagg tcggagggct gggactgtcc cggaaggtgg atgctgaggt      480

gaagaccatt cacagtccca gg                                               502


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

<220>
<223>  Synthetic DNA for SgRNA seq.1 against Glanzmann thrombasthenia

<400>  8
gcagccccca gtccgcctcg                                                   20


<210>  9
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>   Synthetic DNA for SgRNA seq.2 against Glanzmann thrombasthenia

<400>  9
gcagccccca gtacacctcg                                                   20


<210>  10
<211>  500
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic DNA containing portion of human gene encoding for 
       X-linked myotubular myopathy

<400>  10
catggaagta tatattattt cctctctgag aaacttggca tttagaaggc acattgctga       60

attgtattgt cacattttgt gttatatgct ttctcagttt tgtacccatt aattaaaaca      120

aattatcttc atcaatttat tcagcgaata ggtcatggtg ataaaaacca caccgatgct      180

gaccgttctc ctatttttct ccagtttatt gattgtgtgt ggcaaatgtc aaaacaggca      240

aggaatatga gggatgaaaa tacattcaac tcattgttta aattaaacat tttaatataa      300

tgattgggat ttgaccccaa aataatacag ggtagagcta gagatgaagc aagactgggc      360

atgtgtgaat acttgcagaa gccaggtaca tgagcgttca cctttttatt cctgctacta      420

ttgatatatc tgggattttc cctaataaag ttttaaaaat taagtattta actctgatgt      480

ctctactagt ggaattgaca                                                  500


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

<220>
<223>  Synthetic DNA for SgRNA seq.1 against  X-linked myotubular 
       myopathy

<400>  11
ggcaaatgtc aaaaaaggca                                                   20


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

<220>
<223>  Synthetic DNA for SgRNA seq.2 against X-linked myotubular 
       myopathy

<400>  12
ggcaaatgtc aagacaggca                                                   20


<210>  13
<211>  502
<212>  DNA
<213>  Artificial Sequence

<220>
<223>   Synthetic DNA containing portion of human gene encoding for 
       Hereditary Factor VIII deficiency disease(HA)

<400>  13
tgcaggactg atgatagtta attcaggagg cttcaaggca gtgtctgcta ggatttagca       60

caaaggtaga aggcaagcca gggagggaca ctgccctgga gctgaggagg gagaggtgac      120

ggcagtggca ggtgctgcag tggccaccct cagtagaggt cctgtgcctc gcagcccaga      180

acctccatcc tcagggcaat ctggtgcacc caactctggg ggtgaattcg aaggtagcga      240

gtcagtaacg atgggtctag agagttcacc acaggtgtga aggagtcttg atttccctga      300

aaaacctgaa agaggaaaga tagcatttat tggttgtctg acaggacaat ggtcactgca      360

aagcacagac gcttttctca cttctagttg tttccagtag tccaggatta aaccctggaa      420

gatgactaga tgactctgtc ctacgcatct gggacatcta gaattaagag caccctgctt      480

tggaggcctt cactctagct ag                                               502


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

<220>
<223>   Synthetic DNA for SgRNA seq 1 against Hereditary factor VIII 
       deficiency disease(HA)

<400>  14
ggtagcgagt cagtgacgat                                                   20


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

<220>
<223>   Synthetic DNA for SgRNA seq.2 against Hereditary factor VIII 
       deficiency disease(HA)

<400>  15
ggtagcgagt caataacgat                                                   20


<210>  16
<211>  550
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic DNA containing portion of human gene encoding for 
       Sickle Cell Anemia

<400>  16
actcctaagc cagtgccaga agagccaagg acaggtacgg ctgtcatcac ttagacctca       60

ccctgtggag ccacacccta gggttggcca atctactccc aggagcaggg agggcaggag      120

ccagggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctgacac      180

aactgtgttc actagcaacc tcaaacagac accatggtgc atctgactcc tgaggagaag      240

tctgccgtta ctgccctgtg gggcaaggtg aacgtggatg aagttggtgg tgaggccctg      300

ggcaggttgg tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcatgt      360

ggagacagag aagactcttg ggtttctgat aggcactgac tctctctgcc tattggtcta      420

ttttcccacc cttaggctgc tggtggtcta cccttggacc cagaggttct ttgagtcctt      480

tggggatctg tccactcctg atgctgttat gggcaaccct aaggtgaagg ctcatggcaa      540

gaaagtgctc                                                             550


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

<220>
<223>  Synthetic DNA for SgRNA seq.1 against Sickle cell Anemia (SCA)

<400>  17
gtaacggcag acttctcctc                                                   20


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

<220>
<223>  Synthetic DNA for SgRNA seq.2 against Sickle cell Anemia (SCA)

<400>  18
gtaacggcag acttatccac                                                   20


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

<220>
<223>  Synthetic DNA for double mismatch at HBB locus 1

<400>  19
gtaacggcag acttctcaac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 2

<400>  20
gtaacggcag acttctacac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 3

<400>  21
gtaacggcag acttcaccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 4

<400>  22
gtaacggcag acttatccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 5

<400>  23
gtaacggcag actactccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 6

<400>  24
gtaacggcag acatctccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 7

<400>  25
gtaacggcag aattctccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 8

<400>  26
gtaacggcag ccttctccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 9

<400>  27
gtaactgcag acttctccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 10

<400>  28
gtaaaggcag acttctccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 11

<400>  29
gtaccggcag acttctccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 12

<400>  30
gtcacggcag acttctccac                                                   20


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

<220>
<223>   Synthetic DNA for double mismatch at HBB locus 13

<400>  31
gaaacggcag acttctccac                                                   20


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

<220>
<223>  Synthetic WT sgRNA sequence for human HBB locus

<400>  32
gtaacggcag acttctcctc                                                   20


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

<220>
<223>  Synthetic sgRNA sequence with mismatches at PAM proximal 2 and 6 
       positions

<400>  33
gtaacggcag acttatccac                                                   20


<210>  34
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic DNA sequence for HBB mismatched target 1

<400>  34
gaagaaaagt ctgccgttac                                                   20


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

<220>
<223>  Synthetic DNA sequence for HBB mismatched target 2

<400>  35
gaggagaagt ctgccattgc                                                   20


<210>  36
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 1 mismatch  for Glycogen Storage disease type I SNV 
       and 2 mismatch for wild type sequence at 6th and 2nd position

<400>  36
taatacgact cactataggc catgccatgg gaacaacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  37
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR primer for Glycogen storage disease type I

<400>  37
gcataggaga tggaatttgt taaa                                              24


<210>  38
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TCCCTGACTCTGAGGTTTATCC

<400>  38
tccctgactc tgaggtttat cc                                                22


<210>  39
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 1 mismatch  for Glanzman ThRombasthenia SNV and 2 
       mismatch for wild type sequence at 6th and 2nd position

<400>  39
taatacgact cactatagca gcccccagtc cgcctcggtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  40
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR primer for Glanzmann thrombasthenia disease

<400>  40
acagaggccc acagcacac                                                    19


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

<220>
<223>  PCR primer for Glanzmann thrombasthenia disease

<400>  41
cctgggactg tgaatggtct                                                   20


<210>  42
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 1 mismatch  for X-Linked myotubular myopathy SNV and 2
       mismatch for wild type sequence at 6th and 2nd position

<400>  42
taatacgact cactataggc aaatgtcaag acaggcagtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  43
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR primer for X-linked myotubular myopathy disease

<400>  43
cctctctgag aaacttggca tt                                                22


<210>  44
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR primer for X-linked myotubular myopathy disease

<400>  44
tgtcaattcc actagtagag acatca                                            26


<210>  45
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 1 mismatch  for Hemophilia A(FactoR VIII) deficiency 
       SNV and 2 mismatch for wild type sequence at 6th and 2nd position

<400>  45
taatacgact cactataggt agcgagtcag tgacgatgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  46
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR primer for hereditory FactoR VIII deficiency disease

<400>  46
gcaggactga tgatagttaa ttcag                                             25


<210>  47
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR primer for hereditory FactoR VIII deficiency disease

<400>  47
gagtgaaggc ctccaaagca                                                   20


<210>  48
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA sequence for IVC against plasmid containing HBB sequence

<400>  48
taatacgact cactatagta acggcagact tctcctcgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


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

<220>
<223>  PCR primer for 228bp HBB locus amplicon

<400>  49
tctccacatg cccagtttct                                                   20


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

<220>
<223>  PCR primer for 228bp HBB locus amplicon

<400>  50
agtcagggca gagccatcta                                                   20


<210>  51
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 3Rd position of sgRNA

<400>  51
taatacgact cactatagta acggcagact tctcaacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  52
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 4th position of sgRNA

<400>  52
taatacgact cactatagta acggcagact tctacacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  53
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 5th position of sgRNA

<400>  53
taatacgact cactatagta acggcagact tcaccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  54
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 6th position of sgRNA

<400>  54
taatacgact cactatagta acggcagact tatccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  55
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 7th position of sgRNA

<400>  55
taatacgact cactatagta acggcagact actccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  56
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 8th position of sgRNA

<400>  56
taatacgact cactatagta acggcagaca tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  57
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 9th position of sgRNA

<400>  57
taatacgact cactatagta acggcagaat tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  58
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 10th position of sgRNA

<400>  58
taatacgact cactatagta acggcagcct tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  59
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 15th position of sgRNA

<400>  59
taatacgact cactatagta actgcagact tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  60
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 16th position of sgRNA

<400>  60
taatacgact cactatagta aaggcagact tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  61
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 17th position of sgRNA

<400>  61
taatacgact cactatagta ccggcagact tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  62
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 18th position of sgRNA

<400>  62
taatacgact cactatagtc acggcagact tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  63
<211>  72
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  sgRNA with 2 mismatches with Respect to HBB locus sgRNA keeping 
       mismatch at 2nd and 19th position of sgRNA

<400>  63
taatacgact cactatagaa acggcagact tctccacgtt tcagttgcgc cgaaaggcgc       60

tctgtaatca tt                                                           72


<210>  64
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  N gene crRNA sequence feluda

<400>  64
gtggcagtac gtttttgccg                                                   20


<210>  65
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SARS_CoV2 N gene crRNA Forword primer oligo for IVT

<400>  65
taatacgact cactatagtg gcagtacgtt tttgccggtt tcagttgctg aattat           56


<210>  66
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SARS_CoV2 N gene crRNA reverse primer oligo for IVT

<400>  66
ataattcagc aactgaaacc gagggaattt aaggtcttct atagtgagtc gtatta           56


<210>  67
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  S gene crRNA3 feluda

<400>  67
gacattacac catgttcttt                                                   20


<210>  68
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SARS_CoV2 S gene crRNA  Forward primer oligo for IVT feluda

<400>  68
taatacgact cactatagac attacaccat gttctttgtt tcagttgctg aattat           56


<210>  69
<211>  56
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  SARS_CoV2 S gene crRNA reverse primer oligo for IVT feluda

<400>  69
ataattcagc aactgaaacc taattacaaa tgaatctgct atagtgagtc gtatta           56


<210>  70
<211>  39
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  synthetic chimeric guide RNA part 1


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or u

<400>  70
nnnnnnnnnn nnnnnnnnnn guuucaguug cugaauuau                              39


<210>  71
<211>  61
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Syntehtic chimerric guide RNA part 2

<400>  71
guaauuaaug cucuguaauc auuuaaaagu auuuugaacg gaccucuguu ugacacgucu       60

g                                                                       61


<210>  72
<211>  39
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Phosphorothioate modification between positions 1 and 2, and 37 
       and 38

<400>  72
gacauuacac cauguucuuu guuucaguug cugaauuau                              39


<210>  73
<211>  61
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Phosphorothioate modification between positions 1 and 2, and 
       position 59 and 60. FAM modification at 3' position.

<400>  73
guaauuaaug cucuguaauc auuuaaaagu auuuugaacg gaccucuguu ugacacgucu       60

g                                                                       61


<210>  74
<211>  106
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucletide sequence corresponding to sgRNA for a representative 
       locus


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or t

<400>  74
nnnnnnnnnn nnnnnnnnnn agggttttag agctagaaat agcaagttaa aataaggcta       60

gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttt                     106


<210>  75
<211>  106
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  nucleotide sequence corresponding to a representative example of 
       sgRNA

<400>  75
gtaacggcag acttctcctc agggttttag agctagaaat agcaagttaa aataaggcta       60

gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttt                     106


<210>  76
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide sequence corresponding to representative sgRNA for HBB


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or t

<400>  76
nnnnnnnnnn nnnnnnnnnn agggtttcag ttgcgccgaa aggcgctctg taatcattta       60

aaagtatttt gaacggacct ctgtttgaca cgtctg                                 96


<210>  77
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide corresponding to representative sgRNA for HBB locus

<400>  77
gtaacggcag acttctcctc agggtttcag ttgcgccgaa aggcgctctg taatcattta       60

aaagtatttt gaacggacct ctgtttgaca cgtctg                                 96


<210>  78
<211>  99
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Nucleotide corresponding to representative sgRNA sequence for 
       SpCas9

<400>  78
gtaacggcag acttatccac agggttttag agctagaaat agcaagttaa aataaggcta       60

gtccgttatc aacttgaaaa agtggcaccg agtcggtgc                              99


<210>  79
<211>  39
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic representative single guide RNA for FnCas9

<400>  79
gguccaccaa acguaaugcg guuucaguug cugaauuau                              39


<210>  80
<211>  96
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Generic FnCas9 sgRNA sequence for disease detection with one 
       mismatch at positions 16 /17 /18 /19 bp away from PAM


<220>
<221>  misc_feature
<222>  (1)..(20)
<223>  n is a, c, g, or t

<400>  80
nnnnnnnnnn nnnnnnnnnn agggtttcag ttgcgccgaa aggcgctctg taatcattta       60

aaagtatttt gaacggacct ctgtttgaca cgtctg                                 96


