Addgene pEN35-CDKN2A-Ex2-R - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pEN35-CDKN2A-Ex2-R


Map View

AflIII - PciI (8441) pBR322ori-F (8286 .. 8305) SacII (7488) FspI (7330) PvuI (7184) Amp-R (6984 .. 7003) pBRforEco (6575 .. 6593) pGEX 3' (6515 .. 6537) pRS-marker (6396 .. 6415) PsiI (6282) F1ori-F (6221 .. 6242) F1ori-R (6011 .. 6030) PluTI (5854) SfoI (5852) NarI (5851) KasI (5850) SbfI (5847) NotI (5702) BGH-rev (5487 .. 5504) EcoRI (5463) SV40 NLS BsmI (4963) BsaBI * (4821) KflI (4700) hU6-F (1 .. 21) LKO.1 5' (172 .. 191) XbaI (440) Acc65I (446) KpnI (450) SnaBI (713) NcoI (733) BseRI (1124) hybrid intron PaqCI (1229) AgeI (1251) HA SV40 NLS BglII (1573) PspOMI (2678) ApaI (2682) PasI (3092) EcoRV (3176) PmlI (4112) pEN35-CDKN2A-Ex2-R 8446 bp

Sequence View

60 U6 promoter G A G G G C C T A T T T C C C A T G A T T hU6-F g a g g g c c t a t t t c c c a t g a t t c c t t c a t a t t t g c a t a t a c g a t a c a a g g c t g t t a g a g a g c t c c c g g a t a a a g g g t a c t a a g g a a g t a t a a a c g t a t a t g c t a t g t t c c g a c a a t c t c t c

120 U6 promoter a t a a t t g g a a t t a a t t t g a c t g t a a a c a c a a a g a t a t t a g t a c a a a a t a c g t g a c g t a g a t a t t a a c c t t a a t t a a a c t g a c a t t t g t g t t t c t a t a a t c a t g t t t t a t g c a c t g c a t c t

180 U6 promoter G A C T A T C A T LKO.1 5' a a g t a a t a a t t t c t t g g g t a g t t t g c a g t t t t a a a a t t a t g t t t t a a a a t g g a c t a t c a t t t c a t t a t t a a a g a a c c c a t c a a a c g t c a a a a t t t t a a t a c a a a a t t t t a c c t g a t a g t a

240 U6 promoter A T G C T T A C C G T LKO.1 5' a t g c t t a c c g t a a c t t g a a a g t a t t t c g a t t t c t t g g c t t t a t a t a t c t t g t g g a a a g g a t a c g a a t g g c a t t g a a c t t t c a t a a a g c t a a a g a a c c g a a a t a t a t a g a a c a c c t t t c c t

300 U6 promoter c g a a a c a c c g a c c c g t g c a c g a c g c t g c c g t t t a a g a g c t a t g c t g g a a a c a g c a t a g c a g c t t t g t g g c t g g g c a c g t g c t g c g a c g g c a a a t t c t c g a t a c g a c c t t t g t c g t a t c g t

360 a g t t t a a a t a a g g c t a g t c c g t t a t c a a c t t g a a a a a g t g g c a c c g a g t c g g t g c t t t t t t c a a a t t t a t t c c g a t c a g g c a a t a g t t g a a c t t t t t c a c c g t g g c t c a g c c a c g a a a a a

420 t g t t t t a g a g c t a g a a a t a g c a a g t t a a a a t a a g g c t a g t c c g t t t t t a g c g c g t g c g c c a c a a a a t c t c g a t c t t t a t c g t t c a a t t t t a t t c c g a t c a g g c a a a a a t c g c g c a c g c g g

XbaIAcc65IKpnI 480 CMV enhancer a a t t c t g c a g a c a a a t g g c t c t a g a g g t a c c c g t t a c a t a a c t t a c g g t a a a t g g c c c g c t t a a g a c g t c t g t t t a c c g a g a t c t c c a t g g g c a a t g t a t t g a a t g c c a t t t a c c g g g c g

540 CMV enhancer c t g g c t g a c c g c c c a a c g a c c c c c g c c c a t t g a c g t c a a t a g t a a c g c c a a t a g g g a c t t g a c c g a c t g g c g g g t t g c t g g g g g c g g g t a a c t g c a g t t a t c a t t g c g g t t a t c c c t g a a

600 CMV enhancer t c c a t t g a c g t c a a t g g g t g g a g t a t t t a c g g t a a a c t g c c c a c t t g g c a g t a c a t c a a g a g g t a a c t g c a g t t a c c c a c c t c a t a a a t g c c a t t t g a c g g g t g a a c c g t c a t g t a g t t c

660 CMV enhancer t g t a t c a t a t g c c a a g t a c g c c c c c t a t t g a c g t c a a t g a c g g t a a a t g g c c c g c c t g g c a c a t a g t a t a c g g t t c a t g c g g g g g a t a a c t g c a g t t a c t g c c a t t t a c c g g g c g g a c c g

SnaBI 720 CMV enhancer a t t g t g c c c a g t a c a t g a c c t t a t g g g a c t t t c c t a c t t g g c a g t a c a t c t a c g t a t t a g t a a c a c g g g t c a t g t a c t g g a a t a c c c t g a a a g g a t g a a c c g t c a t g t a g a t g c a t a a t c

NcoI 780 CMV enhancer chicken β- actin promoter t c a t c g c t a t t a c c a t g g t c g a g g t g a g c c c c a c g t t c t g c t t c a c t c t c c c c a t c t c c c a g t a g c g a t a a t g g t a c c a g c t c c a c t c g g g g t g c a a g a c g a a g t g a g a g g g g t a g a g g g

840 chicken β- actin promoter c c c c c t c c c c a c c c c c a a t t t t g t a t t t a t t t a t t t t t t a a t t a t t t t g t g c a g c g a t g g g g g g g a g g g g t g g g g g t t a a a a c a t a a a t a a a t a a a a a a t t a a t a a a a c a c g t c g c t a c c

900 chicken β- actin promoter g g g c g g g g g g g g g g g g g g g g c g c g c g c c a g g c g g g g c g g g g c g g g g c g a g g g g c g g g g c g c c c g c c c c c c c c c c c c c c c c g c g c g c g g t c c g c c c c g c c c c g c c c c g c t c c c c g c c c c g c

960 chicken β- actin promoter g g g c g a g g c g g a g a g g t g c g g c g g c a g c c a a t c a g a g c g g c g c g c t c c g a a a g t t t c c t t c c c g c t c c g c c t c t c c a c g c c g c c g t c g g t t a g t c t c g c c g c g c g a g g c t t t c a a a g g a a

1020 chicken β- actin promoter hybrid intron t t a t g g c g a g g c g g c g g c g g c g g c g g c c c t a t a a a a a g c g a a g c g c g c g g c g g g c g g g a g a a t a c c g c t c c g c c g c c g c c g c c g c c g g g a t a t t t t t c g c t t c g c g c g c c g c c c g c c c t c

1080 hybrid intron t c g c t g c g a c g c t g c c t t c g c c c c g t g c c c c g c t c c g c c g c c g c c t c g c g c c g c c c g c c c a g c g a c g c t g c g a c g g a a g c g g g g c a c g g g g c g a g g c g g c g g c g g a g c g c g g c g g g c g g g

BseRI 1140 hybrid intron c g g c t c t g a c t g a c c g c g t t a c t c c c a c a g g t g a g c g g g c g g g a c g g c c c t t c t c c t c c g g c c g a g a c t g a c t g g c g c a a t g a g g g t g t c c a c t c g c c c g c c c t g c c g g g a a g a g g a g g c

1200 hybrid intron g g c t g t a a t t a g c t g a g c a a g a g g t a a g g g t t t a a g g g a t g g t t g g t t g g t g g g g t a t t a c c g a c a t t a a t c g a c t c g t t c t c c a t t c c c a a a t t c c c t a c c a a c c a a c c a c c c c a t a a t

AgeIPaqCI 1260 hybrid intron *TPGTG a t g t t t a a t t a c c t g g a g c a c c t g c c t g a a a t c a c t t t t t t t c a g g t t g g a c c g g t g c c a t a c a a a t t a a t g g a c c t c g t g g a c g g a c t t t a g t g a a a a a a a g t c c a a c c t g g c c a c g g t

1320 1 5 YPYDVPDYA S 130 PKKKRKV E HA SV40 NLS MYPYDVPDYASPKKKRKVE GHVWVINWIVSRRLFLALDF c c a t g t a c c c a t a c g a t g t t c c a g a t t a c g c t t c g c c g a a g a a a a a g c g c a a g g t c g a a g g g t a c a t g g g t a t g c t a c a a g g t c t a a t g c g a a g c g g c t t c t t t t t c g c g t t c c a g c t t c

1380 5 10 15 DKKYSIGLAIGTNSVGWA Cas9 AS 5 DKKYSIGL 10 A 15 IGTNSVGWA Cas9(D10A) ASDKKYSIGLAIGTNSVGWA RGVLLVADAQGDAGVRHAPG c g t c c g a c a a g a a g t a c a g c a t c g g c c t g g c c a t c g g c a c c a a c t c t g t g g g c t g g g c c g g c a g g c t g t t c t t c a t g t c g t a g c c g g a c c g g t a g c c g t g g t t g a g a c a c c c g a c c c g g c

1440 20 25 30 35 VITDEYKVPSKKFKVLGNTD Cas9 20 25 30 35 VITDEYKVPSKKFKVLGNTD Cas9(D10A) VITDEYKVPSKKFKVLGNTD HDGVLVLHGALFELHQAVGV t g a t c a c c g a c g a g t a c a a g g t g c c c a g c a a g a a a t t c a a g g t g c t g g g c a a c a c c g a c c a c t a g t g g c t g c t c a t g t t c c a c g g g t c g t t c t t t a a g t t c c a c g a c c c g t t g t g g c t g g

1500 40 45 50 55 RHSIKKNLIGALLFDSGETA Cas9 40 45 50 55 RHSIKKNLIGALLFDSGETA Cas9(D10A) RHSIKKNLIGALLFDSGETA PVADLLVQDSGQQEVAAFCG g g c a c a g c a t c a a g a a g a a c c t g a t c g g a g c c c t g c t g t t c g a c a g c g g c g a a a c a g c c g c c g t g t c g t a g t t c t t c t t g g a c t a g c c t c g g g a c g a c a a g c t g t c g c c g c t t t g t c g g c

1560 60 65 70 75 EATRLKRTARRRYTRRKNRI Cas9 60 65 70 75 EATRLKRTARRRYTRRKNRI Cas9(D10A) EATRLKRTARRRYTRRKNRI LGGPQLSGGSSSVGSPLVPD a g g c c a c c c g g c t g a a g a g a a c c g c c a g a a g a a g a t a c a c c a g a c g g a a g a a c c g g a t c t t c c g g t g g g c c g a c t t c t c t t g g c g g t c t t c t t c t a t g t g g t c t g c c t t c t t g g c c t a g a

BglII 1620 80 85 90 95 CYLQEIFSNEMAKVDDSFFH Cas9 80 85 90 95 CYLQEIFSNEMAKVDDSFFH Cas9(D10A) CYLQEIFSNEMAKVDDSFFH AIQLLDEAVLHGLHVVAEEV g c t a t c t g c a a g a g a t c t t c a g c a a c g a g a t g g c c a a g g t g g a c g a c a g c t t c t t c c a c a c g a t a g a c g t t c t c t a g a a g t c g t t g c t c t a c c g g t t c c a c c t g c t g t c g a a g a a g g t g t

1680 100 105 110 115 RLEESFLVEEDKKHERHPIF Cas9 100 105 110 115 RLEESFLVEEDKKHERHPIF Cas9(D10A) RLEESFLVEEDKKHERHPIF SQFLGEQHFLILLVLPVGDE g a c t g g a a g a g t c c t t c c t g g t g g a a g a g g a t a a g a a g c a c g a g c g g c a c c c c a t c t t c g c t g a c c t t c t c a g g a a g g a c c a c c t t c t c c t a t t c t t c g t g c t c g c c g t g g g g t a g a a g c

1740 120 125 130 135 GNIVDEVAYHEKYPTIYHLR Cas9 120 125 130 135 GNIVDEVAYHEKYPTIYHLR Cas9(D10A) GNIVDEVAYHEKYPTIYHLR AVDHVLHGVVLLVGGDVVQS g c a a c a t c g t g g a c g a g g t g g c c t a c c a c g a g a a g t a c c c c a c c a t c t a c c a c c t g a g a a c g t t g t a g c a c c t g c t c c a c c g g a t g g t g c t c t t c a t g g g g t g g t a g a t g g t g g a c t c t t

1800 140 145 150 155 KKLVDSTDKADLRLIYLALA Cas9 140 145 150 155 KKLVDSTDKADLRLIYLALA Cas9(D10A) KKLVDSTDKADLRLIYLALA LFQHVAGVLGVQPQDIQGQG a g a a a c t g g t g g a c a g c a c c g a c a a g g c c g a c c t g c g g c t g a t c t a t c t g g c c c t g g c c c t c t t t g a c c a c c t g t c g t g g c t g t t c c g g c t g g a c g c c g a c t a g a t a g a c c g g g a c c g g g

1860 160 165 170 175 HMIKFRGHFLIEGDLNPDNS Cas9 160 165 170 175 HMIKFRGHFLIEGDLNPDNS Cas9(D10A) HMIKFRGHFLIEGDLNPDNS VHDLEPAVEQDLAVQVGVVA a c a t g a t c a a g t t c c g g g g c c a c t t c c t g a t c g a g g g c g a c c t g a a c c c c g a c a a c a g c g t g t a c t a g t t c a a g g c c c c g g t g a a g g a c t a g c t c c c g c t g g a c t t g g g g c t g t t g t c g c

1920 180 185 190 195 DVDKLFIQLVQTYNQLFEEN Cas9 180 185 190 195 DVDKLFIQLVQTYNQLFEEN Cas9(D10A) DVDKLFIQLVQTYNQLFEEN VHVLQEDLQHLGVVLQELFV a c g t g g a c a a g c t g t t c a t c c a g c t g g t g c a g a c c t a c a a c c a g c t g t t c g a g g a a a a c c t g c a c c t g t t c g a c a a g t a g g t c g a c c a c g t c t g g a t g t t g g t c g a c a a g c t c c t t t t g g

1980 200 205 210 215 PINASGVDAKAILSARLSKS Cas9 200 205 210 215 PINASGVDAKAILSARLSKS Cas9(D10A) PINASGVDAKAILSARLSKS GDVGAAHVGLGDQRGSQALA c c a t c a a c g c c a g c g g c g t g g a c g c c a a g g c c a t c c t g t c t g c c a g a c t g a g c a a g a g c a g g t a g t t g c g g t c g c c g c a c c t g c g g t t c c g g t a g g a c a g a c g g t c t g a c t c g t t c t c g t

2040 220 225 230 235 RRLENLIAQLPGEKKNGLFG Cas9 220 225 230 235 RRLENLIAQLPGEKKNGLFG Cas9(D10A) RRLENLIAQLPGEKKNGLFG SPQFIQDGLQGALLLIAQEA g a c g g c t g g a a a a t c t g a t c g c c c a g c t g c c c g g c g a g a a g a a g a a t g g c c t g t t c g g c a c t g c c g a c c t t t t a g a c t a g c g g g t c g a c g g g c c g c t c t t c t t c t t a c c g g a c a a g c c g t

2100 240 245 250 255 NLIALSLGLTPNFKSNFDLA Cas9 240 245 250 255 NLIALSLGLTPNFKSNFDLA Cas9(D10A) NLIALSLGLTPNFKSNFDLA VQNGQAQAQGGVELAVEVQG a c c t g a t t g c c c t g a g c c t g g g c c t g a c c c c c a a c t t c a a g a g c a a c t t c g a c c t g g c c g t g g a c t a a c g g g a c t c g g a c c c g g a c t g g g g g t t g a a g t t c t c g t t g a a g c t g g a c c g g c

2160 260 265 270 275 EDAKLQLSKDTYDDDLDNLL Cas9 260 265 270 275 EDAKLQLSKDTYDDDLDNLL Cas9(D10A) EDAKLQLSKDTYDDDLDNLL LIGFQLQALVGVVVVQVVQQ a g g a t g c c a a a c t g c a g c t g a g c a a g g a c a c c t a c g a c g a c g a c c t g g a c a a c c t g c t g g t c c t a c g g t t t g a c g t c g a c t c g t t c c t g t g g a t g c t g c t g c t g g a c c t g t t g g a c g a c c

2220 280 285 290 295 AQIGDQYADLFLAAKNLSDA Cas9 280 285 290 295 AQIGDQYADLFLAAKNLSDA Cas9(D10A) AQIGDQYADLFLAAKNLSDA GLDAVLVGVQKQGGLVQGVG c c c a g a t c g g c g a c c a g t a c g c c g a c c t g t t t c t g g c c g c c a a g a a c c t g t c c g a c g c c a g g g t c t a g c c g c t g g t c a t g c g g c t g g a c a a a g a c c g g c g g t t c t t g g a c a g g c t g c g g t

2280 300 305 310 315 ILLSDILRVNTEITKAPLSA Cas9 300 305 310 315 ILLSDILRVNTEITKAPLSA Cas9(D10A) ILLSDILRVNTEITKAPLSA DQQAVDQSHVGLDGLGGQAG t c c t g c t g a g c g a c a t c c t g a g a g t g a a c a c c g a g a t c a c c a a g g c c c c c c t g a g c g c c t a g g a c g a c t c g c t g t a g g a c t c t c a c t t g t g g c t c t a g t g g t t c c g g g g g g a c t c g c g g a

2340 320 325 330 335 SMIKRYDEHHQDLTLLKALV Cas9 320 325 330 335 SMIKRYDEHHQDLTLLKALV Cas9(D10A) SMIKRYDEHHQDLTLLKALV RHDLSVVLVVLVQGQQFSEH c t a t g a t c a a g a g a t a c g a c g a g c a c c a c c a g g a c c t g a c c c t g c t g a a a g c t c t c g t g c g a t a c t a g t t c t c t a t g c t g c t c g t g g t g g t c c t g g a c t g g g a c g a c t t t c g a g a g c a c g

2400 340 345 350 355 RQQLPEKYKEIFFDQSKNGY Cas9 340 345 350 355 RQQLPEKYKEIFFDQSKNGY Cas9(D10A) RQQLPEKYKEIFFDQSKNGY PLLQRLLVFLNEEVLALVAV g g c a g c a g c t g c c t g a g a a g t a c a a a g a g a t t t t c t t c g a c c a g a g c a a g a a c g g c t a c g c c g t c g t c g a c g g a c t c t t c a t g t t t c t c t a a a a g a a g c t g g t c t c g t t c t t g c c g a t g c

2460 360 365 370 375 AGYIDGGASQEEFYKFIKPI Cas9 360 365 370 375 AGYIDGGASQEEFYKFIKPI Cas9(D10A) AGYIDGGASQEEFYKFIKPI GAVNVASGALFLEVLEDLGD c c g g c t a c a t t g a c g g c g g a g c c a g c c a g g a a g a g t t c t a c a a g t t c a t c a a g c c c a t c c g g c c g a t g t a a c t g c c g c c t c g g t c g g t c c t t c t c a a g a t g t t c a a g t a g t t c g g g t a g g

2520 380 385 390 395 LEKMDGTEELLVKLNREDLL Cas9 380 385 390 395 LEKMDGTEELLVKLNREDLL Cas9(D10A) LEKMDGTEELLVKLNREDLL QFLHVAGLFQEHLQVSLVQQ t g g a a a a g a t g g a c g g c a c c g a g g a a c t g c t c g t g a a g c t g a a c a g a g a g g a c c t g c t g c a c c t t t t c t a c c t g c c g t g g c t c c t t g a c g a g c a c t t c g a c t t g t c t c t c c t g g a c g a c g

2580 400 405 410 415 RKQRTFDNGSIPHQIHLGEL Cas9 400 405 410 415 RKQRTFDNGSIPHQIHLGEL Cas9(D10A) RKQRTFDNGSIPHQIHLGEL PLLPGEVVAADGVLDVQSLQ g g a a g c a g c g g a c c t t c g a c a a c g g c a g c a t c c c c c a c c a g a t c c a c c t g g g a g a g c t g c c c t t c g t c g c c t g g a a g c t g t t g c c g t c g t a g g g g g t g g t c t a g g t g g a c c c t c t c g a c g

2640 420 425 430 435 HAILRRQEDFYPFLKDNREK Cas9 420 425 430 435 HAILRRQEDFYPFLKDNREK Cas9(D10A) HAILRRQEDFYPFLKDNREK VGNQPPLFIKVWEQLVVPFL a c g c c a t t c t g c g g c g g c a g g a a g a t t t t t a c c c a t t c c t g a a g g a c a a c c g g g a a a a g a t g c g g t a a g a c g c c g c c g t c c t t c t a a a a a t g g g t a a g g a c t t c c t g t t g g c c c t t t t c t

PspOMIApaI 2700 440 445 450 455 IEKILTFRIPYYVGPLARGN Cas9 440 445 450 455 IEKILTFRIPYYVGPLARGN Cas9(D10A) IEKILTFRIPYYVGPLARGN DLLDQGEADGVVHARQGPSV t c g a g a a g a t c c t g a c c t t c c g c a t c c c c t a c t a c g t g g g c c c t c t g g c c a g g g g a a a c a a g c t c t t c t a g g a c t g g a a g g c g t a g g g g a t g a t g c a c c c g g g a g a c c g g t c c c c t t t g t

2760 460 465 470 475 SRFAWMTRKSEETITPWNFE Cas9 460 465 470 475 SRFAWMTRKSEETITPWNFE Cas9(D10A) SRFAWMTRKSEETITPWNFE ASEGPHGSLALFGDGGPVEL g c a g a t t c g c c t g g a t g a c c a g a a a g a g c g a g g a a a c c a t c a c c c c c t g g a a c t t c g a g g c g t c t a a g c g g a c c t a c t g g t c t t t c t c g c t c c t t t g g t a g t g g g g g a c c t t g a a g c t c c

2820 480 485 490 495 EVVDKGASAQSFIERMTNFD Cas9 480 485 490 495 EVVDKGASAQSFIERMTNFD Cas9(D10A) EVVDKGASAQSFIERMTNFD FHHVLASGGLAEDLPHGVEI a a g t g g t g g a c a a g g g c g c t t c c g c c c a g a g c t t c a t c g a g c g g a t g a c c a a c t t c g a t a t t c a c c a c c t g t t c c c g c g a a g g c g g g t c t c g a a g t a g c t c g c c t a c t g g t t g a a g c t a t

2880 500 505 510 515 KNLPNEKVLPKHSLLYEYFT Cas9 500 505 510 515 KNLPNEKVLPKHSLLYEYFT Cas9(D10A) KNLPNEKVLPKHSLLYEYFT LVQGVLLHQGLVAQQVLVEG a g a a c c t g c c c a a c g a g a a g g t g c t g c c c a a g c a c a g c c t g c t g t a c g a g t a c t t c a c c g t c t t g g a c g g g t t g c t c t t c c a c g a c g g g t t c g t g t c g g a c g a c a t g c t c a t g a a g t g g c

2940 520 525 530 535 VYNELTKVKYVTEGMRKPAF Cas9 520 525 530 535 VYNELTKVKYVTEGMRKPAF Cas9(D10A) VYNELTKVKYVTEGMRKPAF HIVLQGFHFVHGLSHSLGGE t g t a t a a c g a g c t g a c c a a a g t g a a a t a c g t g a c c g a g g g a a t g a g a a a g c c c g c c t t c c a c a t a t t g c t c g a c t g g t t t c a c t t t a t g c a c t g g c t c c c t t a c t c t t t c g g g c g g a a g g

3000 540 545 550 555 LSGEQKKAIVDLLFKTNRKV Cas9 540 545 550 555 LSGEQKKAIVDLLFKTNRKV Cas9(D10A) LSGEQKKAIVDLLFKTNRKV QAALLFLGDHVQQELGVPFH t g a g c g g c g a g c a g a a a a a g g c c a t c g t g g a c c t g c t g t t c a a g a c c a a c c g g a a a g t g a a c t c g c c g c t c g t c t t t t t c c g g t a g c a c c t g g a c g a c a a g t t c t g g t t g g c c t t t c a c t

3060 560 565 570 575 TVKQLKEDYFKKIECFDSVE Cas9 560 565 570 575 TVKQLKEDYFKKIECFDSVE Cas9(D10A) TVKQLKEDYFKKIECFDSVE GHLLQFLVVELFDLAEVGHF c c g t g a a g c a g c t g a a a g a g g a c t a c t t c a a g a a a a t c g a g t g c t t c g a c t c c g t g g a a a g g c a c t t c g t c g a c t t t c t c c t g a t g a a g t t c t t t t a g c t c a c g a a g c t g a g g c a c c t t t

PasI 3120 580 585 590 595 ISGVEDRFNASLGTYHDLLK Cas9 580 585 590 595 ISGVEDRFNASLGTYHDLLK Cas9(D10A) ISGVEDRFNASLGTYHDLLK DGAHFIPEVGGQACVVIQQF t c t c c g g c g t g g a a g a t c g g t t c a a c g c c t c c c t g g g c a c a t a c c a c g a t c t g c t g a a a a a g a g g c c g c a c c t t c t a g c c a a g t t g c g g a g g g a c c c g t g t a t g g t g c t a g a c g a c t t t t

EcoRV 3180 600 605 610 615 IIKDKDFLDNEENEDILEDI Cas9 600 605 610 615 IIKDKDFLDNEENEDILEDI Cas9(D10A) IIKDKDFLDNEENEDILEDI NDLVLVEQVILFVLVNQFID t t a t c a a g g a c a a g g a c t t c c t g g a c a a t g a g g a a a a c g a g g a c a t t c t g g a a g a t a t c g a a t a g t t c c t g t t c c t g a a g g a c c t g t t a c t c c t t t t g c t c c t g t a a g a c c t t c t a t a g c

3240 620 625 630 635 VLTLTLFEDREMIEERLKTY Cas9 620 625 630 635 VLTLTLFEDREMIEERLKTY Cas9(D10A) VLTLTLFEDREMIEERLKTY HQGQCQKLVSLHDLFPQFGI *A t g c t g a c c c t g a c a c t g t t t g a g g a c a g a g a g a t g a t c g a g g a a c g g c t g a a a a c c t a t g a c g a c t g g g a c t g t g a c a a a c t c c t g t c t c t c t a c t a g c t c c t t g c c g a c t t t t g g a t a c

3300 640 645 650 655 AHLFDDKVMKQLKRRRYTGW Cas9 640 645 650 655 AHLFDDKVMKQLKRRRYTGW Cas9(D10A) AHLFDDKVMKQLKRRRYTGW GVQEVVFHHLLQLPPSVGAP WRNSSLTIFCSFRRLYVPQP c c c a c c t g t t c g a c g a c a a a g t g a t g a a g c a g c t g a a g c g g c g g a g a t a c a c c g g c t g g g g g g t g g a c a a g c t g c t g t t t c a c t a c t t c g t c g a c t t c g c c g c c t c t a t g t g g c c g a c c c

3360 660 665 670 675 GRLSRKLINGIRDKQSGKTI Cas9 660 665 670 675 GRLSRKLINGIRDKQSGKTI Cas9(D10A) GRLSRKLINGIRDKQSGKTI APQAPLQDVADPVLLGALCD LSLRFSILPMRSLCDPLVIR g c a g g c t g a g c c g g a a g c t g a t c a a c g g c a t c c g g g a c a a g c a g t c c g g c a a g a c a a t c c c g t c c g a c t c g g c c t t c g a c t a g t t g c c g t a g g c c c t g t t c g t c a g g c c g t t c t g t t a g g

3420 680 685 690 695 LDFLKSDGFANRNFMQLIHD Cas9 680 685 690 695 LDFLKSDGFANRNFMQLIHD Cas9(D10A) LDFLKSDGFANRNFMQLIHD QIEQLGVAEGVSVEHLQDVV SKRFDSPKALLFKMCSIWSS t g g a t t t c c t g a a g t c c g a c g g c t t c g c c a a c a g a a a c t t c a t g c a g c t g a t c c a c g a c g a c c t a a a g g a c t t c a g g c t g c c g a a g c g g t t g t c t t t g a a g t a c g t c g a c t a g g t g c t g c

3480 700 705 710 715 DSLTFKEDIQKAQVSGQGDS Cas9 700 705 710 715 DSLTFKEDIQKAQVSGQGDS Cas9(D10A) DSLTFKEDIQKAQVSGQGDS VAQGKFLVDLFGLHGALAIA LRVKLSSMWFAWTDPWPSLR a c a g c c t g a c c t t t a a a g a g g a c a t c c a g a a a g c c c a g g t g t c c g g c c a g g g c g a t a g c c t g t c g g a c t g g a a a t t t c t c c t g t a g g t c t t t c g g g t c c a c a g g c c g g t c c c g c t a t c g g

3540 720 725 730 735 LHEHIANLAGSPAIKKGILQ Cas9 720 725 730 735 LHEHIANLAGSPAIKKGILQ Cas9(D10A) LHEHIANLAGSPAIKKGILQ QVLVNGIQGAAGGNLLADQL CSCMALRAPLGAMLFPMRCV t g c a c g a g c a c a t t g c c a a t c t g g c c g g c a g c c c c g c c a t t a a g a a g g g c a t c c t g c a g a a c g t g c t c g t g t a a c g g t t a g a c c g g c c g t c g g g g c g g t a a t t c t t c c c g t a g g a c g t c t

3600 740 745 750 755 TVKVVDELVKVMGRHKPENI Cas9 740 745 750 755 TVKVVDELVKVMGRHKPENI Cas9(D10A) TVKVVDELVKVMGRHKPENI CHLHHVLEHFHHAPVLGLVD TFTTSSSTFTIPRCLGSFMT c a g t g a a g g t g g t g g a c g a g c t c g t g a a a g t g a t g g g c c g g c a c a a g c c c g a g a a c a t c g g t c a c t t c c a c c a c c t g c t c g a g c a c t t t c a c t a c c c g g c c g t g t t c g g g c t c t t g t a g c

3660 760 765 770 775 VIEMARENQTTQKGQKNSRE Cas9 760 765 770 775 VIEMARENQTTQKGQKNSRE Cas9(D10A) VIEMARENQTTQKGQKNSRE HDFHGSLVLGGLLSLLVAAL ISIALSFWVVWFPCFFLRSL t g a t c g a a a t g g c c a g a g a g a a c c a g a c c a c c c a g a a g g g a c a g a a g a a c a g c c g c g a g a a c t a g c t t t a c c g g t c t c t c t t g g t c t g g t g g g t c t t c c c t g t c t t c t t g t c g g c g c t c t

3720 780 785 790 795 RMKRIEEGIKELGSQILKEH Cas9 780 785 790 795 RMKRIEEGIKELGSQILKEH Cas9(D10A) RMKRIEEGIKELGSQILKEH SHLPDFLADFLQAALDQFFV IFRISSPM g a a t g a a g c g g a t c g a a g a g g g c a t c a a a g a g c t g g g c a g c c a g a t c c t g a a a g a a c a c c c t t a c t t c g c c t a g c t t c t c c c g t a g t t t c t c g a c c c g t c g g t c t a g g a c t t t c t t g t g g

3780 800 805 810 815 PVENTQLQNEKLYLYYLQNG Cas9 800 805 810 815 PVENTQLQNEKLYLYYLQNG Cas9(D10A) PVENTQLQNEKLYLYYLQNG GHFVGLQLVLLQVQVVQLIP c c g t g g a a a a c a c c c a g c t g c a g a a c g a g a a g c t g t a c c t g t a c t a c c t g c a g a a t g g g c g g c a c c t t t t g t g g g t c g a c g t c t t g c t c t t c g a c a t g g a c a t g a t g g a c g t c t t a c c c g

3840 820 825 830 835 RDMYVDQELDINRLSDYDVD Cas9 820 825 830 835 RDMYVDQELDINRLSDYDVD Cas9(D10A) RDMYVDQELDINRLSDYDVD PIHVHVLFQVDVPQGVVIHV g g g a t a t g t a c g t g g a c c a g g a a c t g g a c a t c a a c c g g c t g t c c g a c t a c g a t g t g g a c c c c c t a t a c a t g c a c c t g g t c c t t g a c c t g t a g t t g g c c g a c a g g c t g a t g c t a c a c c t g g

3900 840 845 850 855 HIVPQSFLKDDSIDNKVLTR Cas9 840 845 850 855 HIVPQSFLKDDSIDNKVLTR Cas9(D10A) HIVPQSFLKDDSIDNKVLTR M a t a t c g t g c c t c a g a g c t t t c t g a a g g a c g a c t c c a t c g a c a a c a a g g t g c t g a c c a g a a t a t a g c a c g g a g t c t c g a a a g a c t t c c t g c t g a g g t a g c t g t t g t t c c a c g a c t g g t c t t

3960 860 865 870 875 SDKNRGKSDNVPSEEVVKKM Cas9 860 865 870 875 SDKNRGKSDNVPSEEVVKKM Cas9(D10A) SDKNRGKSDNVPSEEVVKKM g c g a c a a g a a c c g g g g c a a g a g c g a c a a c g t g c c c t c c g a a g a g g t c g t g a a g a a g a t g a c g c t g t t c t t g g c c c c g t t c t c g c t g t t g c a c g g g a g g c t t c t c c a g c a c t t c t t c t a c t

4020 880 885 890 895 KNYWRQLLNAKLITQRKFDN Cas9 880 885 890 895 KNYWRQLLNAKLITQRKFDN Cas9(D10A) KNYWRQLLNAKLITQRKFDN a g a a c t a c t g g c g g c a g c t g c t g a a c g c c a a g c t g a t t a c c c a g a g a a a g t t c g a c a a t c t c t t g a t g a c c g c c g t c g a c g a c t t g c g g t t c g a c t a a t g g g t c t c t t t c a a g c t g t t a g

4080 900 905 910 915 LTKAERGGLSELDKAGFIKR Cas9 900 905 910 915 LTKAERGGLSELDKAGFIKR Cas9(D10A) LTKAERGGLSELDKAGFIKR t g a c c a a g g c c g a g a g a g g c g g c c t g a g c g a a c t g g a t a a g g c c g g c t t c a t c a a g a g a c a c t g g t t c c g g c t c t c t c c g c c g g a c t c g c t t g a c c t a t t c c g g c c g a a g t a g t t c t c t g

PmlI 4140 920 925 930 935 QLVETRQITKHVAQILDSRM Cas9 920 925 930 935 QLVETRQITKHVAQILDSRM Cas9(D10A) QLVETRQITKHVAQILDSRM a g c t g g t g g a a a c c c g g c a g a t c a c a a a g c a c g t g g c a c a g a t c c t g g a c t c c c g g a t g a t c g a c c a c c t t t g g g c c g t c t a g t g t t t c g t g c a c c g t g t c t a g g a c c t g a g g g c c t a c t

4200 940 945 950 955 NTKYDENDKLIREVKVITLK Cas9 940 945 950 955 NTKYDENDKLIREVKVITLK Cas9(D10A) NTKYDENDKLIREVKVITLK a c a c t a a g t a c g a c g a g a a t g a c a a g c t g a t c c g g g a a g t g a a a g t g a t c a c c c t g a a g t t g t g a t t c a t g c t g c t c t t a c t g t t c g a c t a g g c c c t t c a c t t t c a c t a g t g g g a c t t c a

4260 960 965 970 975 SKLVSDFRKDFQFYKVREIN Cas9 960 965 970 975 SKLVSDFRKDFQFYKVREIN Cas9(D10A) SKLVSDFRKDFQFYKVREIN c c a a g c t g g t g t c c g a t t t c c g g a a g g a t t t c c a g t t t t a c a a a g t g c g c g a g a t c a a c a g g t t c g a c c a c a g g c t a a a g g c c t t c c t a a a g g t c a a a a t g t t t c a c g c g c t c t a g t t g t

4320 980 985 990 995 NYHHAHDAYLNAVVGTALIK Cas9 980 985 990 995 NYHHAHDAYLNAVVGTALIK Cas9(D10A) NYHHAHDAYLNAVVGTALIK a c t a c c a c c a c g c c c a c g a c g c c t a c c t g a a c g c c g t c g t g g g a a c c g c c c t g a t c a a a a t g a t g g t g g t g c g g g t g c t g c g g a t g g a c t t g c g g c a g c a c c c t t g g c g g g a c t a g t t t t

4380 1000 1005 1010 1015 KYPKLESEFVYGDYKVYDVR Cas9 1000 1005 1010 1015 KYPKLESEFVYGDYKVYDVR Cas9(D10A) KYPKLESEFVYGDYKVYDVR a g t a c c c t a a g c t g g a a a g c g a g t t c g t g t a c g g c g a c t a c a a g g t g t a c g a c g t g c g g a t c a t g g g a t t c g a c c t t t c g c t c a a g c a c a t g c c g c t g a t g t t c c a c a t g c t g c a c g c c t

4440 1020 1025 1030 1035 KMIAKSEQEIGKATAKYFFY Cas9 1020 1025 1030 1035 KMIAKSEQEIGKATAKYFFY Cas9(D10A) KMIAKSEQEIGKATAKYFFY a g a t g a t c g c c a a g a g c g a g c a g g a a a t c g g c a a g g c t a c c g c c a a g t a c t t c t t c t a c a t c t a c t a g c g g t t c t c g c t c g t c c t t t a g c c g t t c c g a t g g c g g t t c a t g a a g a a g a t g t

4500 1040 1045 1050 1055 SNIMNFFKTEITLANGEIRK Cas9 1040 1045 1050 1055 SNIMNFFKTEITLANGEIRK Cas9(D10A) SNIMNFFKTEITLANGEIRK g c a a c a t c a t g a a c t t t t t c a a g a c c g a g a t t a c c c t g g c c a a c g g c g a g a t c c g g a a g c c g t t g t a g t a c t t g a a a a a g t t c t g g c t c t a a t g g g a c c g g t t g c c g c t c t a g g c c t t c g

4560 1060 1065 1070 1075 RPLIETNGETGEIVWDKGRD Cas9 1060 1065 1070 1075 RPLIETNGETGEIVWDKGRD Cas9(D10A) RPLIETNGETGEIVWDKGRD g g c c t c t g a t c g a g a c a a a c g g c g a a a c c g g g g a g a t c g t g t g g g a t a a g g g c c g g g a t t c c g g a g a c t a g c t c t g t t t g c c g c t t t g g c c c c t c t a g c a c a c c c t a t t c c c g g c c c t a a

4620 1080 1085 1090 1095 FATVRKVLSMPQVNIVKKTE Cas9 1080 1085 1090 1095 FATVRKVLSMPQVNIVKKTE Cas9(D10A) FATVRKVLSMPQVNIVKKTE t t g c c a c c g t g c g g a a a g t g c t g a g c a t g c c c c a a g t g a a t a t c g t g a a a a a g a c c g a g g a a c g g t g g c a c g c c t t t c a c g a c t c g t a c g g g g t t c a c t t a t a g c a c t t t t t c t g g c t c c

4680 1100 1105 1110 1115 VQTGGFSKESILPKRNSDKL Cas9 1100 1105 1110 1115 VQTGGFSKESILPKRNSDKL Cas9(D10A) VQTGGFSKESILPKRNSDKL t g c a g a c a g g c g g c t t c a g c a a a g a g t c t a t c c t g c c c a a g a g g a a c a g c g a t a a g c t g a a c g t c t g t c c g c c g a a g t c g t t t c t c a g a t a g g a c g g g t t c t c c t t g t c g c t a t t c g a c t

KflI 4740 1120 1125 1130 1135 IARKKDWDPKKYGGFDSPTV Cas9 1120 1125 1130 1135 IARKKDWDPKKYGGFDSPTV Cas9(D10A) IARKKDWDPKKYGGFDSPTV t c g c c a g a a a g a a g g a c t g g g a c c c t a a g a a g t a c g g c g g c t t c g a c a g c c c c a c c g t g g a g c g g t c t t t c t t c c t g a c c c t g g g a t t c t t c a t g c c g c c g a a g c t g t c g g g g t g g c a c c

4800 1140 1145 1150 1155 AYSVLVVAKVEKGKSKKLKS Cas9 1140 1145 1150 1155 AYSVLVVAKVEKGKSKKLKS Cas9(D10A) AYSVLVVAKVEKGKSKKLKS c c t a t t c t g t g c t g g t g g t g g c c a a a g t g g a a a a g g g c a a g t c c a a g a a a c t g a a g a g t g g g a t a a g a c a c g a c c a c c a c c g g t t t c a c c t t t t c c c g t t c a g g t t c t t t g a c t t c t c a c

BsaBI* 4860 1160 1165 1170 1175 VKELLGITIMERSSFEKNPI Cas9 1160 1165 1170 1175 VKELLGITIMERSSFEKNPI Cas9(D10A) VKELLGITIMERSSFEKNPI t g a a a g a g c t g c t g g g g a t c a c c a t c a t g g a a a g a a g c a g c t t c g a g a a g a a t c c c a t c g a c t t t c t c g a c g a c c c c t a g t g g t a g t a c c t t t c t t c g t c g a a g c t c t t c t t a g g g t a g c

4920 1180 1185 1190 1195 DFLEAKGYKEVKKDLIIKLP Cas9 1180 1185 1190 1195 DFLEAKGYKEVKKDLIIKLP Cas9(D10A) DFLEAKGYKEVKKDLIIKLP a c t t t c t g g a a g c c a a g g g c t a c a a a g a a g t g a a a a a g g a c c t g a t c a t c a a g c t g c c t a t g a a a g a c c t t c g g t t c c c g a t g t t t c t t c a c t t t t t c c t g g a c t a g t a g t t c g a c g g a t

BsmI 4980 1200 1205 1210 1215 KYSLFELENGRKRMLASAGE Cas9 1200 1205 1210 1215 KYSLFELENGRKRMLASAGE Cas9(D10A) KYSLFELENGRKRMLASAGE a g t a c t c c c t g t t c g a g c t g g a a a a c g g c c g g a a g a g a a t g c t g g c c t c t g c c g g c g a a c t c a t g a g g g a c a a g c t c g a c c t t t t g c c g g c c t t c t c t t a c g a c c g g a g a c g g c c g c t t g

5040 1220 1225 1230 1235 LQKGNELALPSKYVNFLYLA Cas9 1220 1225 1230 1235 LQKGNELALPSKYVNFLYLA Cas9(D10A) LQKGNELALPSKYVNFLYLA t g c a g a a g g g a a a c g a a c t g g c c c t g c c c t c c a a a t a t g t g a a c t t c c t g t a c c t g g c c a a c g t c t t c c c t t t g c t t g a c c g g g a c g g g a g g t t t a t a c a c t t g a a g g a c a t g g a c c g g t

5100 1240 1245 1250 1255 SHYEKLKGSPEDNEQKQLFV Cas9 1240 1245 1250 1255 SHYEKLKGSPEDNEQKQLFV Cas9(D10A) SHYEKLKGSPEDNEQKQLFV g c c a c t a t g a g a a g c t g a a g g g c t c c c c c g a g g a t a a t g a g c a g a a a c a g c t g t t t g t g g c g g t g a t a c t c t t c g a c t t c c c g a g g g g g c t c c t a t t a c t c g t c t t t g t c g a c a a a c a c c

5160 1260 1265 1270 1275 EQHKHYLDEIIEQISEFSKR Cas9 1260 1265 1270 1275 EQHKHYLDEIIEQISEFSKR Cas9(D10A) EQHKHYLDEIIEQISEFSKR a a c a g c a c a a g c a c t a c c t g g a c g a g a t c a t c g a g c a g a t c a g c g a g t t c t c c a a g a g a g t t g t c g t g t t c g t g a t g g a c c t g c t c t a g t a g c t c g t c t a g t c g c t c a a g a g g t t c t c t c

5220 1280 1285 1290 1295 VILADANLDKVLSAYNKHRD Cas9 1280 1285 1290 1295 VILADANLDKVLSAYNKHRD Cas9(D10A) VILADANLDKVLSAYNKHRD t g a t c c t g g c c g a c g c t a a t c t g g a c a a a g t g c t g t c c g c c t a c a a c a a g c a c c g g g a t a a c t a g g a c c g g c t g c g a t t a g a c c t g t t t c a c g a c a g g c g g a t g t t g t t c g t g g c c c t a t

5280 1300 1305 1310 1315 KPIREQAENIIHLFTLTNLG Cas9 1300 1305 1310 1315 KPIREQAENIIHLFTLTNLG Cas9(D10A) KPIREQAENIIHLFTLTNLG a g c c c a t c a g a g a g c a g g c c g a g a a t a t c a t c c a c c t g t t t a c c c t g a c c a a t c t g g g a g t c g g g t a g t c t c t c g t c c g g c t c t t a t a g t a g g t g g a c a a a t g g g a c t g g t t a g a c c c t c

5340 1320 1325 1330 1335 APAAFKYFDTTIDRKRYTST Cas9 1320 1325 1330 1335 APAAFKYFDTTIDRKRYTST Cas9(D10A) APAAFKYFDTTIDRKRYTST c c c c t g c c g c c t t c a a g t a c t t t g a c a c c a c c a t c g a c c g g a a g a g g t a c a c c a g c a c c a g g g g a c g g c g g a a g t t c a t g a a a c t g t g g t g g t a g c t g g c c t t c t c c a t g t g g t c g t g g t

5400 1340 1345 1350 1355 KEVLDATLIHQSITGLYETR Cas9 1340 1345 1350 1355 KEVLDATLIHQSITGLYETR Cas9(D10A) KEVLDATLIHQSITGLYETR a a g a g g t g c t g g a c g c c a c c c t g a t c c a c c a g a g c a t c a c c g g c c t g t a c g a g a c a c g g a t t c t c c a c g a c c t g c g g t g g g a c t a g g t g g t c t c g t a g t g g c c g g a c a t g c t c t g t g c c t

5460 1360 1365 IDLSQLGGD (in frame with Cas9) SPKKKRKVEAS Cas9 1360 1365 IDLSQLGGD S 130 PKKKRKV EAS Cas9(D10A) SV40 NLS IDLSQLGGDSPKKKRKVEAS t c g a c c t g t c t c a g c t g g g a g g c g a c a g c c c c a a g a a g a a g a g a a a g g t g g a g g c c a g c t a g c t g g a c a g a g t c g a c c c t c c g c t g t c g g g g t t c t t c t t c t c t t t c c a c c t c c g g t c g a

EcoRI 5520 * bGH poly(A) signal * G G A G C T G A C A C G G A A G A T BGH-rev * a a g a a t t c c t a g a g c t c g c t g a t c a g c c t c g a c t g t g c c t t c t a g t t g c c a g c c a t c t g t t t c t t a a g g a t c t c g a g c g a c t a g t c g g a g c t g a c a c g g a a g a t c a a c g g t c g g t a g a c a

5580 bGH poly(A) signal t g t t t g c c c c t c c c c c g t g c c t t c c t t g a c c c t g g a a g g t g c c a c t c c c a c t g t c c t t t c a c a a a c g g g g a g g g g g c a c g g a a g g a a c t g g g a c c t t c c a c g g t g a g g g t g a c a g g a a a g

5640 bGH poly(A) signal c t a a t a a a a t g a g g a a a t t g c a t c g c a t t g t c t g a g t a g g t g t c a t t c t a t t c t g g g g g g g a t t a t t t t a c t c c t t t a a c g t a g c g t a a c a g a c t c a t c c a c a g t a a g a t a a g a c c c c c c

5700 bGH poly(A) signal t g g g g t g g g g c a g g a c a g c a a g g g g g a g g a t t g g g a a g a g a a t a g c a g g c a t g c t g g g g a a c c c c a c c c c g t c c t g t c g t t c c c c c t c c t a a c c c t t c t c t t a t c g t c c g t a c g a c c c c t

NotI 5760 AAV2 ITR AAV2 ITR g c g g c c g c a g g a a c c c c t a g t g a t g g a g t t g g c c a c t c c c t c t c t g c g c g c t c g c t c g c t c g c c g g c g t c c t t g g g g a t c a c t a c c t c a a c c g g t g a g g g a g a g a c g c g c g a g c g a g c g a

5820 AAV2 ITR AAV2 ITR c a c t g a g g c c g g g c g a c c a a a g g t c g c c c g a c g c c c g g g c t t t g c c c g g g c g g c c t c a g t g t g a c t c c g g c c c g c t g g t t t c c a g c g g g c t g c g g g c c c g a a a c g g g c c c g c c g g a g t c a

SbfIKasINarISfoIPluTI 5880 AAV2 ITR AAV2 ITR g a g c g a g c g a g c g c g c a g c t g c c t g c a g g g g c g c c t g a t g c g g t a t t t t c t c c t t a c g c a c t c g c t c g c t c g c g c g t c g a c g g a c g t c c c c g c g g a c t a c g c c a t a a a a g a g g a a t g c g t

5940 f1 ori t c t g t g c g g t a t t t c a c a c c g c a t a c g t c a a a g c a a c c a t a g t a c g c g c c c t g t a g c g g c a g a c a c g c c a t a a a g t g t g g c g t a t g c a g t t t c g t t g g t a t c a t g c g c g g g a c a t c g c c g

6000 f1 ori g c a t t a a g c g c g g c g g g t g t g g t g g t t a c g c g c a g c g t g a c c g c t a c a c t t g c c a g c g c c c g t a a t t c g c g c c g c c c a c a c c a c c a a t g c g c g t c g c a c t g g c g a t g t g a a c g g t c g c g g

6060 f1 ori G A G G A A A G C G A A A G A A G G G A F1ori-R c t a g c g c c c g c t c c t t t c g c t t t c t t c c c t t c c t t t c t c g c c a c g t t c g c c g g c t t t c c c g a t c g c g g g c g a g g a a a g c g a a a g a a g g g a a g g a a a g a g c g g t g c a a g c g g c c g a a a g g g

6120 f1 ori c g t c a a g c t c t a a a t c g g g g g c t c c c t t t a g g g t t c c g a t t t a g t g c t t t a c g g c a c c t c g c a g t t c g a g a t t t a g c c c c c g a g g g a a a t c c c a a g g c t a a a t c a c g a a a t g c c g t g g a g

6180 f1 ori g a c c c c a a a a a a c t t g a t t t g g g t g a t g g t t c a c g t a g t g g g c c a t c g c c c t g a t a g a c g c t g g g g t t t t t t g a a c t a a a c c c a c t a c c a a g t g c a t c a c c c g g t a g c g g g a c t a t c t g c

6240 f1 ori G T G G A C T C T T G T T C C A A A C T F1ori-F g t t t t t c g c c c t t t g a c g t t g g a g t c c a c g t t c t t t a a t a g t g g a c t c t t g t t c c a a a c t c a a a a a g c g g g a a a c t g c a a c c t c a g g t g c a a g a a a t t a t c a c c t g a g a a c a a g g t t t g a

PsiI 6300 f1 ori G G F1ori-F g g a a c a a c a c t c a a c c c t a t c t c g g g c t a t t c t t t t g a t t t a t a a g g g a t t t t g c c g a t t c c t t g t t g t g a g t t g g g a t a g a g c c c g a t a a g a a a a c t a a a t a t t c c c t a a a a c g g c t a a

6360 f1 ori t c g g c c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a a a a t t t a a c g c g a a t t t t a a c a a a a g c c g g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t t t t t a a a t t g c g c t t a a a a t t g t t t

6420 f1 ori A T G T T A G A C G A G A C T A C G G C pRS-marker a t a t t a a c g t t t a c a a t t t t a t g g t g c a c t c t c a g t a c a a t c t g c t c t g a t g c c g c a t a g t a t a a t t g c a a a t g t t a a a a t a c c a c g t g a g a g t c a t g t t a g a c g a g a c t a c g g c g t a t c

6480 t t a a g c c a g c c c c g a c a c c c g c c a a c a c c c g c t g a c g c g c c c t g a c g g g c t t g t c t g c t c a a t t c g g t c g g g g c t g t g g g c g g t t g t g g g c g a c t g c g c g g g a c t g c c c g a a c a g a c g a g

6540 C C G G G A G C T G C A T G T G T C A G A G G pGEX 3' c c g g c a t c c g c t t a c a g a c a a g c t g t g a c c g t c t c c g g g a g c t g c a t g t g t c a g a g g t t t g g c c g t a g g c g a a t g t c t g t t c g a c a c t g g c a g a g g c c c t c g a c g t a c a c a g t c t c c a a a

6600 C G G A G C A C T A T G C G G A T A A pBRforEco t c a c c g t c a t c a c c g a a a c g c g c g a g a c g a a a g g g c c t c g t g a t a c g c c t a t t t t t a t a g a g t g g c a g t a g t g g c t t t g c g c g c t c t g c t t t c c c g g a g c a c t a t g c g g a t a a a a a t a t c

6660 g t t a a t g t c a t g a t a a t a a t g g t t t c t t a g a c g t c a g g t g g c a c t t t t c g g g g a a a t g t g c a a t t a c a g t a c t a t t a t t a c c a a a g a a t c t g c a g t c c a c c g t g a a a a g c c c c t t t a c a c

6720 AmpR promoter c g c g g a a c c c c t a t t t g t t t a t t t t t c t a a a t a c a t t c a a a t a t g t a t c c g c t c a t g a g a g c g c c t t g g g g a t a a a c a a a t a a a a a g a t t t a t g t a a g t t t a t a c a t a g g c g a g t a c t c t

6780 1 5 signal sequence MSIQH AmpR promoter AmpR MSIQH c a a t a a c c c t g a t a a a t g c t t c a a t a a t a t t g a a a a a g g a a g a g t a t g a g t a t t c a a c a t g t t a t t g g g a c t a t t t a c g a a g t t a t t a t a a c t t t t t c c t t c t c a t a c t c a t a a g t t g t a

6840 10 15 20 signal sequence FRVALIPFFAAFCLPVFA 25 HP AmpR FRVALIPFFAAFCLPVFAHP t t c c g t g t c g c c c t t a t t c c c t t t t t t g c g g c a t t t t g c c t t c c t g t t t t t g c t c a c c c a a a g g c a c a g c g g g a a t a a g g g a a a a a a c g c c g t a a a a c g g a a g g a c a a a a a c g a g t g g g t

6900 30 35 40 45 ETLVKVKDAEDQLGARVGYI AmpR ETLVKVKDAEDQLGARVGYI g a a a c g c t g g t g a a a g t a a a a g a t g c t g a a g a t c a g t t g g g t g c a c g a g t g g g t t a c a t c c t t t g c g a c c a c t t t c a t t t t c t a c g a c t t c t a g t c a a c c c a c g t g c t c a c c c a a t g t a g

6960 50 55 60 65 ELDLNSGKILESFRPEERFP AmpR ELDLNSGKILESFRPEERFP g a a c t g g a t c t c a a c a g c g g t a a g a t c c t t g a g a g t t t t c g c c c c g a a g a a c g t t t t c c a c t t g a c c t a g a g t t g t c g c c a t t c t a g g a a c t c t c a a a a g c g g g g c t t c t t g c a a a a g g t

7020 70 75 80 85 MMSTFKVLLCGAVLSRIDAG AmpR C G A T A C A C C G C G C C A T A A T A Amp-R MMSTFKVLLCGAVLSRIDAG a t g a t g a g c a c t t t t a a a g t t c t g c t a t g t g g c g c g g t a t t a t c c c g t a t t g a c g c c g g g t a c t a c t c g t g a a a a t t t c a a g a c g a t a c a c c g c g c c a t a a t a g g g c a t a a c t g c g g c c c

7080 90 95 100 105 QEQLGRRIHYSQNDLVEYSP AmpR QEQLGRRIHYSQNDLVEYSP c a a g a g c a a c t c g g t c g c c g c a t a c a c t a t t c t c a g a a t g a c t t g g t t g a g t a c t c a c c a g t t c t c g t t g a g c c a g c g g c g t a t g t g a t a a g a g t c t t a c t g a a c c a a c t c a t g a g t g g t

7140 110 115 120 125 VTEKHLTDGMTVRELCSAAI AmpR VTEKHLTDGMTVRELCSAAI g t c a c a g a a a a g c a t c t t a c g g a t g g c a t g a c a g t a a g a g a a t t a t g c a g t g c t g c c a t a c a g t g t c t t t t c g t a g a a t g c c t a c c g t a c t g t c a t t c t c t t a a t a c g t c a c g a c g g t a t

PvuI 7200 130 135 140 145 TMSDNTAANLLLTTIGGPKE AmpR TMSDNTAANLLLTTIGGPKE a c c a t g a g t g a t a a c a c t g c g g c c a a c t t a c t t c t g a c a a c g a t c g g a g g a c c g a a g g a g t g g t a c t c a c t a t t g t g a c g c c g g t t g a a t g a a g a c t g t t g c t a g c c t c c t g g c t t c c t c

7260 150 155 160 165 LTAFLHNMGDHVTRLDRWEP AmpR LTAFLHNMGDHVTRLDRWEP c t a a c c g c t t t t t t g c a c a a c a t g g g g g a t c a t g t a a c t c g c c t t g a t c g t t g g g a a c c g g a t t g g c g a a a a a a c g t g t t g t a c c c c c t a g t a c a t t g a g c g g a a c t a g c a a c c c t t g g c

7320 170 175 180 185 ELNEAIPNDERDTTMPVAMA AmpR ELNEAIPNDERDTTMPVAMA g a g c t g a a t g a a g c c a t a c c a a a c g a c g a g c g t g a c a c c a c g a t g c c t g t a g c a a t g g c a c t c g a c t t a c t t c g g t a t g g t t t g c t g c t c g c a c t g t g g t g c t a c g g a c a t c g t t a c c g t

FspI 7380 190 195 200 205 TTLRKLLTGELLTLASRQQL AmpR TTLRKLLTGELLTLASRQQL a c a a c g t t g c g c a a a c t a t t a a c t g g c g a a c t a c t t a c t c t a g c t t c c c g g c a a c a a t t a t g t t g c a a c g c g t t t g a t a a t t g a c c g c t t g a t g a a t g a g a t c g a a g g g c c g t t g t t a a t

7440 210 215 220 225 IDWMEADKVAGPLLRSALPA AmpR IDWMEADKVAGPLLRSALPA a t a g a c t g g a t g g a g g c g g a t a a a g t t g c a g g a c c a c t t c t g c g c t c g g c c c t t c c g g c t t a t c t g a c c t a c c t c c g c c t a t t t c a a c g t c c t g g t g a a g a c g c g a g c c g g g a a g g c c g a

SacII 7500 230 235 240 245 GWFIADKSGAGERGSRGIIA AmpR GWFIADKSGAGERGSRGIIA g g c t g g t t t a t t g c t g a t a a a t c t g g a g c c g g t g a g c g t g g a a g c c g c g g t a t c a t t g c a c c g a c c a a a t a a c g a c t a t t t a g a c c t c g g c c a c t c g c a c c t t c g g c g c c a t a g t a a c g t

7560 250 255 260 265 ALGPDGKPSRIVVIYTTGSQ AmpR ALGPDGKPSRIVVIYTTGSQ g c a c t g g g g c c a g a t g g t a a g c c c t c c c g t a t c g t a g t t a t c t a c a c g a c g g g g a g t c a g c g t g a c c c c g g t c t a c c a t t c g g g a g g g c a t a g c a t c a a t a g a t g t g c t g c c c c t c a g t c

7620 270 275 280 285 ATMDERNRQIAEIGASLIKH AmpR ATMDERNRQIAEIGASLIKH g c a a c t a t g g a t g a a c g a a a t a g a c a g a t c g c t g a g a t a g g t g c c t c a c t g a t t a a g c a t c g t t g a t a c c t a c t t g c t t t a t c t g t c t a g c g a c t c t a t c c a c g g a g t g a c t a a t t c g t a

7680 W* AmpR W* t g g t a a c t g t c a g a c c a a g t t t a c t c a t a t a t a c t t t a g a t t g a t t t a a a a c t t c a t t t t a c c a t t g a c a g t c t g g t t c a a a t g a g t a t a t a t g a a a t c t a a c t a a a t t t t g a a g t a a a a

7740 t a a t t t a a a a g g a t c t a g g t g a a g a t c c t t t t t g a t a a t c t c a t g a c c a a a a t c c c t t a a a t t a a a t t t t c c t a g a t c c a c t t c t a g g a a a a a c t a t t a g a g t a c t g g t t t t a g g g a a t t

7800 ori c g t g a g t t t t c g t t c c a c t g a g c g t c a g a c c c c g t a g a a a a g a t c a a a g g a t c t t c t t g a g c a c t c a a a a g c a a g g t g a c t c g c a g t c t g g g g c a t c t t t t c t a g t t t c c t a g a a g a a c t

7860 ori g a t c c t t t t t t t c t g c g c g t a a t c t g c t g c t t g c a a a c a a a a a a a c c a c c g c t a c c a g c g c t a g g a a a a a a a g a c g c g c a t t a g a c g a c g a a c g t t t g t t t t t t t g g t g g c g a t g g t c g c

7920 ori g t g g t t t g t t t g c c g g a t c a a g a g c t a c c a a c t c t t t t t c c g a a g g t a a c t g g c t t c a g c c a c c a a a c a a a c g g c c t a g t t c t c g a t g g t t g a g a a a a a g g c t t c c a t t g a c c g a a g t c g

7980 ori a g a g c g c a g a t a c c a a a t a c t g t c c t t c t a g t g t a g c c g t a g t t a g g c c a c c a c t t c a a g t c t c g c g t c t a t g g t t t a t g a c a g g a a g a t c a c a t c g g c a t c a a t c c g g t g g t g a a g t t c

8040 ori a a c t c t g t a g c a c c g c c t a c a t a c c t c g c t c t g c t a a t c c t g t t a c c a g t g g c t g c t g c c t t g a g a c a t c g t g g c g g a t g t a t g g a g c g a g a c g a t t a g g a c a a t g g t c a c c g a c g a c g g

8100 ori a g t g g c g a t a a g t c g t g t c t t a c c g g g t t g g a c t c a a g a c g a t a g t t a c c g g a t a a g g c g t c a c c g c t a t t c a g c a c a g a a t g g c c c a a c c t g a g t t c t g c t a t c a a t g g c c t a t t c c g c

8160 ori c a g c g g t c g g g c t g a a c g g g g g g t t c g t g c a c a c a g c c c a g c t t g g a g c g a a c g a c c t a c g t c g c c a g c c c g a c t t g c c c c c c a a g c a c g t g t g t c g g g t c g a a c c t c g c t t g c t g g a t g

8220 ori a c c g a a c t g a g a t a c c t a c a g c g t g a g c t a t g a g a a a g c g c c a c g c t t c c c g a a g g g a g a t g g c t t g a c t c t a t g g a t g t c g c a c t c g a t a c t c t t t c g c g g t g c g a a g g g c t t c c c t c t

8280 ori a a g g c g g a c a g g t a t c c g g t a a g c g g c a g g g t c g g a a c a g g a g a g c g c a c g a g g g a g c t t t t c c g c c t g t c c a t a g g c c a t t c g c c g t c c c a g c c t t g t c c t c t c g c g t g c t c c c t c g a a

8340 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F c c a g g g g g a a a c g c c t g g t a t c t t t a t a g t c c t g t c g g g t t t c g c c a c c t c t g a c t t g a g g g t c c c c c t t t g c g g a c c a t a g a a a t a t c a g g a c a g c c c a a a g c g g t g g a g a c t g a a c t c

8400 ori c g t c g a t t t t t g t g a t g c t c g t c a g g g g g g c g g a g c c t a t g g a a a a a c g c c a g c a a c g c g g c a g c t a a a a a c a c t a c g a g c a g t c c c c c c g c c t c g g a t a c c t t t t t g c g g t c g t t g c g c

AflIIIPciI 8446 g c c t t t t t a c g g t t c c t g g c c t t t t g c t g g c c t t t t g c t c a c a t g t c g g a a a a a t g c c a a g g a c c g g a a a a c g a c c g g a a a a c g a g t g t a c a

Restriction Enzymes

Instructions: By default, all cutters are shown. Filter on number of cut sites or search by enzyme name.

Filter

Features

Primers

BLAST

BLAST (Basic Local Alignment Search Tool) finds regions of similarity between biological sequences. Click on the buttons below to submit a BLAST search to NCBI. The results will appear in a new window. See your recent BLAST results on NCBI's website.

  • Nucleotide-Nucleotide BLAST (BLASTN)

  • Translated Nucleotide-Protein BLAST (BLASTX)

  • Sequence alignment using BLAST (BLAST2)

Sequence Analyzer Guide

Map

Displays a graphical map based on nucleotide sequence data labeled with restriction enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site)

To select a portion of sequence, click one location on the plasmid and then a second location to display the sequence between the two locations.

Sequence

Displays both strands of base paired nucleotide sequences with annotated enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site).

To select a portion of sequence, click one location on the sequence and then a second location to display the sequence between the two locations.

Enzymes

List of restriction enzymes that can cut a given nucleotide sequence. Table lists enzyme name and the sequence location of the cut.

Features

List of common features detected in a given nucleotide sequence. Table lists feature name, location, size, color used to indicate its position on the map, and direction (if relevant).

Primers

List of commonly used primers detected in a given nucleotide sequence. Table lists primer name, sequence, length, binding site location, and direction.

BLAST

Use Basic Local Alignment Search Tool (BLAST) via the NCBI website to determine similarity between a given sequence and nucleotide (BLASTN) or protein (BLASTX) sequence databases. Additionally, align a custom nucleotide sequence against a given sequence using BLAST2.

File Downloads

GenBank

File contains the nucleotide sequence and annotated features in GenBank flat file format. Open the file with a text editor or plasmid mapping software to view the sequence.

SnapGene

File contains the nucleotide sequence and enhanced annotations from SnapGene Server. Open the file with SnapGene software or the free Viewer to view the plasmid map, sequence, and perform additional sequence analysis.