Addgene PX458_AHR_2 Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: PX458_AHR_2 Sequencing Result


Map View

hU6-F (15 .. 35) AflIII - PciI (9) pBR322ori-F (9145 .. 9164) SacII (8347) FspI (8189) PvuI (8043) Amp-R (7843 .. 7862) pBRforEco (7434 .. 7452) pGEX 3' (7374 .. 7396) pRS-marker (7255 .. 7274) PsiI (7141) F1ori-F (7080 .. 7101) F1ori-R (6870 .. 6889) PluTI (6713) SfoI (6711) NarI (6710) KasI (6709) SbfI (6706) NotI (6561) BGH-rev (6346 .. 6363) BsrGI (6311) EGFP-C (6255 .. 6276) EXFP-R (5908 .. 5927) T2A FseI (5516) BsmI (5024) BsaBI * (4882) LKO.1 5' (186 .. 205) XbaI (445) Acc65I (451) KpnI (455) SnaBI (718) chicken β-actin promoter PaqCI (1233) AgeI (1255) SV40 NLS BglII (1634) PspOMI (2739) ApaI (2743) PasI (3153) EcoRV (3237) PmlI (4173) KflI (4761) PX458_AHR_2 9291 bp

Sequence View

AflIIIPciI 5ʹ 3ʹ 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 t t t t g c t c a c a t g t 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 a a a c g a g t g t a c a 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

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

180 U6 promoter a t a c g t g a c g t a g a 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 t a t g c a c t g c a t c 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

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

300 U6 promoter gRNA scaffold t c t t g t g g a a a g g a c g a a a c a c c g c a a a a a c c a g g g t c a a a a t t g t t t t a g a g c t a g a a a a g a a c a c c t t t c c t g c t t t g t g g c g t t t t t g g t c c c a g t t t t a a c a a a a t c t c g a t c t t t

360 gRNA scaffold 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 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 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 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

420 t t t t t 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 a a a a a 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

XbaIAcc65IKpnI 480 CMV enhancer g c g c c 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 g c g g 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

540 CMV enhancer c c c g c 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 g g c g 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

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

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

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

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

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

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

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

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

1080 chicken β- actin promoter hybrid intron g g g a g t c g c t g c g 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 c c c t c a g c g a c g c 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

1140 hybrid intron g c c c 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 c g g 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

1200 hybrid intron t c c g 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 g g c 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

AgeIPaqCI 1260 hybrid intron *TPGT a t t a 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 t a a t 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

1320 1 5 10 15 DYKDHDGDYKDHDIDYK 3xFLAG Kozak sequence MDYKDHDGDYKDHDIDYK GGHVILVVVSVVLIMINIVF g c c a c c a t g g a c t a t a a g g a c c a c g a c g g a g a c t a c a a g g a t c a t g a t a t t g a t t a c a a a c g g t g g t a c c t g a t a t t c c t g g t g c t g c c t c t g a t g t t c c t a g t a c t a t a a c t a a t g t t t

1380 20 DDDDK MA 130 PKKKRKV (in frame with SV40 NLS) GIHGVP 3xFLAG SV40 NLS DDDDKMAPKKKRKVGIHGVP VIVILHGWLLLPLDTDVSDW g a c g a t g a c g a t a a g a t g g c c c c a a a g a a g a a g c g g a a g g t c g g t a t c c a c g g a g t c c c a c t g c t a c t g c t a t t c t a c c g g g g t t t c t t c t t c g c c t t c c a g c c a t a g g t g c c t c a g g g t

1440 5 10 15 DKKYSIGLDIGTNSVGWA Cas9 AA 5 10 15 DKKYSIGLDIGTNSVGWA Cas9 AADKKYSIGLDIGTNSVGWA CGVLLVADAQVDAGVRHAPG g c a g 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 a 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 c g t c 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 t 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

1500 20 25 30 35 VITDEYKVPSKKFKVLGNTD Cas9 20 25 30 35 VITDEYKVPSKKFKVLGNTD Cas9 VITDEYKVPSKKFKVLGNTD HDGVLVLHGALFELHQAVGV g 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

1560 40 45 50 55 RHSIKKNLIGALLFDSGETA Cas9 40 45 50 55 RHSIKKNLIGALLFDSGETA Cas9 RHSIKKNLIGALLFDSGETA PVADLLVQDSGQQEVAAFCG c 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

1620 60 65 70 75 EATRLKRTARRRYTRRKNRI Cas9 60 65 70 75 EATRLKRTARRRYTRRKNRI Cas9 EATRLKRTARRRYTRRKNRI LGGPQLSGGSSSVGSPLVPD g 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 c 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

BglII 1680 80 85 90 95 CYLQEIFSNEMAKVDDSFFH Cas9 80 85 90 95 CYLQEIFSNEMAKVDDSFFH Cas9 CYLQEIFSNEMAKVDDSFFH AIQLLDEAVLHGLHVVAEEV t 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

1740 100 105 110 115 RLEESFLVEEDKKHERHPIF Cas9 100 105 110 115 RLEESFLVEEDKKHERHPIF Cas9 RLEESFLVEEDKKHERHPIF SQFLGEQHFLILLVLPVGDE a 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 t 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

1800 120 125 130 135 GNIVDEVAYHEKYPTIYHLR Cas9 120 125 130 135 GNIVDEVAYHEKYPTIYHLR Cas9 GNIVDEVAYHEKYPTIYHLR AVDHVLHGVVLLVGGDVVQS g 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 c 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

1860 140 145 150 155 KKLVDSTDKADLRLIYLALA Cas9 140 145 150 155 KKLVDSTDKADLRLIYLALA Cas9 KKLVDSTDKADLRLIYLALA LFQHVAGVLGVQPQDIQGQG a 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 t 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

1920 160 165 170 175 HMIKFRGHFLIEGDLNPDNS Cas9 160 165 170 175 HMIKFRGHFLIEGDLNPDNS Cas9 HMIKFRGHFLIEGDLNPDNS VHDLEPAVEQDLAVQVGVVA c 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

1980 180 185 190 195 DVDKLFIQLVQTYNQLFEEN Cas9 180 185 190 195 DVDKLFIQLVQTYNQLFEEN Cas9 DVDKLFIQLVQTYNQLFEEN VHVLQEDLQHLGVVLQELFV g 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

2040 200 205 210 215 PINASGVDAKAILSARLSKS Cas9 200 205 210 215 PINASGVDAKAILSARLSKS Cas9 PINASGVDAKAILSARLSKS GDVGAAHVGLGDQRGSQALA c 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 g 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

2100 220 225 230 235 RRLENLIAQLPGEKKNGLFG Cas9 220 225 230 235 RRLENLIAQLPGEKKNGLFG Cas9 RRLENLIAQLPGEKKNGLFG SPQFIQDGLQGALLLIAQES a 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 a t 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 t

2160 240 245 250 255 NLIALSLGLTPNFKSNFDLA Cas9 240 245 250 255 NLIALSLGLTPNFKSNFDLA Cas9 NLIALSLGLTPNFKSNFDLA VQNGQAQAQGGVELAVEVQG a 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 t 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

2220 260 265 270 275 EDAKLQLSKDTYDDDLDNLL Cas9 260 265 270 275 EDAKLQLSKDTYDDDLDNLL Cas9 EDAKLQLSKDTYDDDLDNLL LIGFQLQALVGVVVVQVVQQ g 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 c 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

2280 280 285 290 295 AQIGDQYADLFLAAKNLSDA Cas9 280 285 290 295 AQIGDQYADLFLAAKNLSDA Cas9 AQIGDQYADLFLAAKNLSDA GLDAVLVGVQKQGGLVQGVG g 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 c 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

2340 300 305 310 315 ILLSDILRVNTEITKAPLSA Cas9 300 305 310 315 ILLSDILRVNTEITKAPLSA Cas9 ILLSDILRVNTEITKAPLSA DQQAVDQSHVGLDGLGGQAG a 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

2400 320 325 330 335 SMIKRYDEHHQDLTLLKALV Cas9 320 325 330 335 SMIKRYDEHHQDLTLLKALV Cas9 SMIKRYDEHHQDLTLLKALV RHDLSVVLVVLVQGQQFSEH t 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 a 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

2460 340 345 350 355 RQQLPEKYKEIFFDQSKNGY Cas9 340 345 350 355 RQQLPEKYKEIFFDQSKNGY Cas9 RQQLPEKYKEIFFDQSKNGY PLLQRLLVFLNEEVLALVAV c 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

2520 360 365 370 375 AGYIDGGASQEEFYKFIKPI Cas9 360 365 370 375 AGYIDGGASQEEFYKFIKPI Cas9 AGYIDGGASQEEFYKFIKPI GAVNVASGALFLEVLEDLGD g 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

2580 380 385 390 395 LEKMDGTEELLVKLNREDLL Cas9 380 385 390 395 LEKMDGTEELLVKLNREDLL Cas9 LEKMDGTEELLVKLNREDLL QFLHVAGLFQEHLQVSLVQQ c 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 g 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

2640 400 405 410 415 RKQRTFDNGSIPHQIHLGEL Cas9 400 405 410 415 RKQRTFDNGSIPHQIHLGEL Cas9 RKQRTFDNGSIPHQIHLGEL PLLPGEVVAADGVLDVQSLQ c 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 g 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

2700 420 425 430 435 HAILRRQEDFYPFLKDNREK Cas9 420 425 430 435 HAILRRQEDFYPFLKDNREK Cas9 HAILRRQEDFYPFLKDNREK VGNQPPLFIKVWEQLVVPFL c 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 g 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

PspOMIApaI 2760 440 445 450 455 IEKILTFRIPYYVGPLARGN Cas9 440 445 450 455 IEKILTFRIPYYVGPLARGN Cas9 IEKILTFRIPYYVGPLARGN DLLDQGEADGVVHARQGPSV a 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 t 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

2820 460 465 470 475 SRFAWMTRKSEETITPWNFE Cas9 460 465 470 475 SRFAWMTRKSEETITPWNFE Cas9 SRFAWMTRKSEETITPWNFE ASEGPHGSLALFGDGGPVEL a 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 t 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

2880 480 485 490 495 EVVDKGASAQSFIERMTNFD Cas9 480 485 490 495 EVVDKGASAQSFIERMTNFD Cas9 EVVDKGASAQSFIERMTNFD FHHVLASGGLAEDLPHGVEI g 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 c 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

2940 500 505 510 515 KNLPNEKVLPKHSLLYEYFT Cas9 500 505 510 515 KNLPNEKVLPKHSLLYEYFT Cas9 KNLPNEKVLPKHSLLYEYFT LVQGVLLHQGLVAQQVLVEG a 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 t 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

3000 520 525 530 535 VYNELTKVKYVTEGMRKPAF Cas9 520 525 530 535 VYNELTKVKYVTEGMRKPAF Cas9 VYNELTKVKYVTEGMRKPAF HIVLQGFHFVHGLSHSLGGE g 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

3060 540 545 550 555 LSGEQKKAIVDLLFKTNRKV Cas9 540 545 550 555 LSGEQKKAIVDLLFKTNRKV Cas9 LSGEQKKAIVDLLFKTNRKV QAALLFLGDHVQQELGVPFH c 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 g 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

3120 560 565 570 575 TVKQLKEDYFKKIECFDSVE Cas9 560 565 570 575 TVKQLKEDYFKKIECFDSVE Cas9 TVKQLKEDYFKKIECFDSVE GHLLQFLVVELFDLAEVGHF a 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 t 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

PasI 3180 580 585 590 595 ISGVEDRFNASLGTYHDLLK Cas9 580 585 590 595 ISGVEDRFNASLGTYHDLLK Cas9 ISGVEDRFNASLGTYHDLLK DGAHFIPEVGGQACVVIQQF a 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 t 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

EcoRV 3240 600 605 610 615 IIKDKDFLDNEENEDILEDI Cas9 600 605 610 615 IIKDKDFLDNEENEDILEDI Cas9 IIKDKDFLDNEENEDILEDI NDLVLVEQVILFVLVNQFID a 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 t 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

3300 620 625 630 635 VLTLTLFEDREMIEERLKTY Cas9 620 625 630 635 VLTLTLFEDREMIEERLKTY Cas9 VLTLTLFEDREMIEERLKTY * HQGQCQKLVSLHDLFPQFGI g 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 c 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

3360 640 645 650 655 AHLFDDKVMKQLKRRRYTGW Cas9 640 645 650 655 AHLFDDKVMKQLKRRRYTGW Cas9 AHLFDDKVMKQLKRRRYTGW AWRNSSLTIFCSFRRLYVPQ GVQEVVFHHLLQLPPSVGAP g 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 c 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

3420 660 665 670 675 GRLSRKLINGIRDKQSGKTI Cas9 660 665 670 675 GRLSRKLINGIRDKQSGKTI Cas9 GRLSRKLINGIRDKQSGKTI PLSLRFSILPMRSLCDPLVI APQAPLQDVADPVLLGALCD g 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

3480 680 685 690 695 LDFLKSDGFANRNFMQLIHD Cas9 680 685 690 695 LDFLKSDGFANRNFMQLIHD Cas9 LDFLKSDGFANRNFMQLIHD RSKRFDSPKALLFKMCSIWS QIEQLGVAEGVSVEHLQDVV c 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

3540 700 705 710 715 DSLTFKEDIQKAQVSGQGDS Cas9 700 705 710 715 DSLTFKEDIQKAQVSGQGDS Cas9 DSLTFKEDIQKAQVSGQGDS SLRVKLSSMWFAWTDPWPSL VAQGKFLVDLFGLHGALAIA g 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

3600 720 725 730 735 LHEHIANLAGSPAIKKGILQ Cas9 720 725 730 735 LHEHIANLAGSPAIKKGILQ Cas9 LHEHIANLAGSPAIKKGILQ RCSCMALRAPLGAMLFPMRC QVLVNGIQGAAGGNLLADQL c 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 g 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

3660 740 745 750 755 TVKVVDELVKVMGRHKPENI Cas9 740 745 750 755 TVKVVDELVKVMGRHKPENI Cas9 TVKVVDELVKVMGRHKPENI VTFTTSSSTFTIPRCLGSFM CHLHHVLEHFHHAPVLGLVD a 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 t 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

3720 760 765 770 775 VIEMARENQTTQKGQKNSRE Cas9 760 765 770 775 VIEMARENQTTQKGQKNSRE Cas9 VIEMARENQTTQKGQKNSRE TISIALSFWVVWFPCFFLRS HDFHGSLVLGGLLSLLVAAL g 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 c 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

3780 780 785 790 795 RMKRIEEGIKELGSQILKEH Cas9 780 785 790 795 RMKRIEEGIKELGSQILKEH Cas9 RMKRIEEGIKELGSQILKEH LIFRISSPM SHLPDFLADFLQAALDQFFV a 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 t 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

3840 800 805 810 815 PVENTQLQNEKLYLYYLQNG Cas9 800 805 810 815 PVENTQLQNEKLYLYYLQNG Cas9 PVENTQLQNEKLYLYYLQNG GHFVGLQLVLLQVQVVQLIP c 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 g 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

3900 820 825 830 835 RDMYVDQELDINRLSDYDVD Cas9 820 825 830 835 RDMYVDQELDINRLSDYDVD Cas9 RDMYVDQELDINRLSDYDVD PIHVHVLFQVDVPQGVVIHV c 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 g 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

3960 840 845 850 855 HIVPQSFLKDDSIDNKVLTR Cas9 840 845 850 855 HIVPQSFLKDDSIDNKVLTR Cas9 HIVPQSFLKDDSIDNKVLTR M c 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 g 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

4020 860 865 870 875 SDKNRGKSDNVPSEEVVKKM Cas9 860 865 870 875 SDKNRGKSDNVPSEEVVKKM Cas9 SDKNRGKSDNVPSEEVVKKM a 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 t 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

4080 880 885 890 895 KNYWRQLLNAKLITQRKFDN Cas9 880 885 890 895 KNYWRQLLNAKLITQRKFDN Cas9 KNYWRQLLNAKLITQRKFDN a 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 t 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

4140 900 905 910 915 LTKAERGGLSELDKAGFIKR Cas9 900 905 910 915 LTKAERGGLSELDKAGFIKR Cas9 LTKAERGGLSELDKAGFIKR c 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 g 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

PmlI 4200 920 925 930 935 QLVETRQITKHVAQILDSRM Cas9 920 925 930 935 QLVETRQITKHVAQILDSRM Cas9 QLVETRQITKHVAQILDSRM c 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 g 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

4260 940 945 950 955 NTKYDENDKLIREVKVITLK Cas9 940 945 950 955 NTKYDENDKLIREVKVITLK Cas9 NTKYDENDKLIREVKVITLK a 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

4320 960 965 970 975 SKLVSDFRKDFQFYKVREIN Cas9 960 965 970 975 SKLVSDFRKDFQFYKVREIN Cas9 SKLVSDFRKDFQFYKVREIN t 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

4380 980 985 990 995 NYHHAHDAYLNAVVGTALIK Cas9 980 985 990 995 NYHHAHDAYLNAVVGTALIK Cas9 NYHHAHDAYLNAVVGTALIK a 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 t 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

4440 1000 1005 1010 1015 KYPKLESEFVYGDYKVYDVR Cas9 1000 1005 1010 1015 KYPKLESEFVYGDYKVYDVR Cas9 KYPKLESEFVYGDYKVYDVR a 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 t 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

4500 1020 1025 1030 1035 KMIAKSEQEIGKATAKYFFY Cas9 1020 1025 1030 1035 KMIAKSEQEIGKATAKYFFY Cas9 KMIAKSEQEIGKATAKYFFY a 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 t 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

4560 1040 1045 1050 1055 SNIMNFFKTEITLANGEIRK Cas9 1040 1045 1050 1055 SNIMNFFKTEITLANGEIRK Cas9 SNIMNFFKTEITLANGEIRK a 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 t 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

4620 1060 1065 1070 1075 RPLIETNGETGEIVWDKGRD Cas9 1060 1065 1070 1075 RPLIETNGETGEIVWDKGRD Cas9 RPLIETNGETGEIVWDKGRD c 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 g 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

4680 1080 1085 1090 1095 FATVRKVLSMPQVNIVKKTE Cas9 1080 1085 1090 1095 FATVRKVLSMPQVNIVKKTE Cas9 FATVRKVLSMPQVNIVKKTE t 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 a 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

4740 1100 1105 1110 1115 VQTGGFSKESILPKRNSDKL Cas9 1100 1105 1110 1115 VQTGGFSKESILPKRNSDKL Cas9 VQTGGFSKESILPKRNSDKL g 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 c 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

KflI 4800 1120 1125 1130 1135 IARKKDWDPKKYGGFDSPTV Cas9 1120 1125 1130 1135 IARKKDWDPKKYGGFDSPTV Cas9 IARKKDWDPKKYGGFDSPTV a 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 t 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

4860 1140 1145 1150 1155 AYSVLVVAKVEKGKSKKLKS Cas9 1140 1145 1150 1155 AYSVLVVAKVEKGKSKKLKS Cas9 AYSVLVVAKVEKGKSKKLKS g 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 c 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

BsaBI* 4920 1160 1165 1170 1175 VKELLGITIMERSSFEKNPI Cas9 1160 1165 1170 1175 VKELLGITIMERSSFEKNPI Cas9 VKELLGITIMERSSFEKNPI g 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 c 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

4980 1180 1185 1190 1195 DFLEAKGYKEVKKDLIIKLP Cas9 1180 1185 1190 1195 DFLEAKGYKEVKKDLIIKLP Cas9 DFLEAKGYKEVKKDLIIKLP g 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 c 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

BsmI 5040 1200 1205 1210 1215 KYSLFELENGRKRMLASAGE Cas9 1200 1205 1210 1215 KYSLFELENGRKRMLASAGE Cas9 KYSLFELENGRKRMLASAGE a 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 t 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

5100 1220 1225 1230 1235 LQKGNELALPSKYVNFLYLA Cas9 1220 1225 1230 1235 LQKGNELALPSKYVNFLYLA Cas9 LQKGNELALPSKYVNFLYLA c 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 g 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

5160 1240 1245 1250 1255 SHYEKLKGSPEDNEQKQLFV Cas9 1240 1245 1250 1255 SHYEKLKGSPEDNEQKQLFV Cas9 SHYEKLKGSPEDNEQKQLFV a 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 t 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

5220 1260 1265 1270 1275 EQHKHYLDEIIEQISEFSKR Cas9 1260 1265 1270 1275 EQHKHYLDEIIEQISEFSKR Cas9 EQHKHYLDEIIEQISEFSKR g 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 c 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

5280 1280 1285 1290 1295 VILADANLDKVLSAYNKHRD Cas9 1280 1285 1290 1295 VILADANLDKVLSAYNKHRD Cas9 VILADANLDKVLSAYNKHRD g 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 c 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

5340 1300 1305 1310 1315 KPIREQAENIIHLFTLTNLG Cas9 1300 1305 1310 1315 KPIREQAENIIHLFTLTNLG Cas9 KPIREQAENIIHLFTLTNLG a 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 t 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

5400 1320 1325 1330 1335 APAAFKYFDTTIDRKRYTST Cas9 1320 1325 1330 1335 APAAFKYFDTTIDRKRYTST Cas9 APAAFKYFDTTIDRKRYTST g 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 c 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

5460 1340 1345 1350 1355 KEVLDATLIHQSITGLYETR Cas9 1340 1345 1350 1355 KEVLDATLIHQSITGLYETR Cas9 KEVLDATLIHQSITGLYETR a 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 t 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

FseI 5520 1360 1365 IDLSQLGGD (in frame with Cas9) KRPAATKKAGQ Cas9 1360 1365 IDLSQLGGD 1 5 10 KRPAATKKAGQ Cas9 nucleoplasmin NLS IDLSQLGGDKRPAATKKAGQ a 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 a a a g g c c g g c g g c c a c g a a a a a g g c c g g c c a g 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 t t t c c g g c c g c c g g t g c t t t t t c c g g c c g g t c

5580 (in frame with Cas9) AKKKKEFGSGEGRGSLLTCG 15 AKKKK EFGSG 140 145 EGRGSLLTCG nucleoplasmin NLS T2A AKKKKEFGSGEGRGSLLTCG g c a a a a a a g a a a a a g g a a t t c g g c a g t g g a g a g g g c a g a g g a a g t c t g c t a a c a t g c g g t c g t t t t t t c t t t t t c c t t a a g c c g t c a c c t c t c c c g t c t c c t t c a g a c g a t t g t a c g c c a

5640 (in frame with Cas9) DVEENPGP 1a V 5 10 SKGEELFTGVV EGFP 150 155 DVEENPGP 1a V 5 10 SKGEELFTGVV T2A EGFP DVEENPGPVSKGEELFTGVV g a c g t c g a g g a g a a t c c t g g c c c a g t g a g c a a g g g c g a g g a g c t g t t c a c c g g g g t g g t g c t g c a g c t c c t c t t a g g a c c g g g t c a c t c g t t c c c g c t c c t c g a c a a g t g g c c c c a c c a c

5700 15 20 25 30 PILVELDGDVNGHKFSVSGE EGFP 15 20 25 30 PILVELDGDVNGHKFSVSGE EGFP PILVELDGDVNGHKFSVSGE c c c a t c c t g g t c g a g c t g g a c g g c g a c g t a a a c g g c c a c a a g t t c a g c g t g t c c g g c g a g g g g t a g g a c c a g c t c g a c c t g c c g c t g c a t t t g c c g g t g t t c a a g t c g c a c a g g c c g c t c

5760 35 40 45 50 GEGDATYGKLTLKFICTTGK EGFP 35 40 45 50 GEGDATYGKLTLKFICTTGK EGFP GEGDATYGKLTLKFICTTGK g g c g a g g g c g a t g c c a c c t a c g g c a a g c t g a c c c t g a a g t t c a t c t g c a c c a c c g g c a a g c c g c t c c c g c t a c g g t g g a t g c c g t t c g a c t g g g a c t t c a a g t a g a c g t g g t g g c c g t t c

5820 55 60 65 70 LPVPWPTLVTTLTYGVQCFS EGFP 55 60 65 70 LPVPWPTLVTTLTYGVQCFS EGFP LPVPWPTLVTTLTYGVQCFS c t g c c c g t g c c c t g g c c c a c c c t c g t g a c c a c c c t g a c c t a c g g c g t g c a g t g c t t c a g c g a c g g g c a c g g g a c c g g g t g g g a g c a c t g g t g g g a c t g g a t g c c g c a c g t c a c g a a g t c g

5880 75 80 85 90 RYPDHMKQHDFFKSAMPEGY EGFP 75 80 85 90 RYPDHMKQHDFFKSAMPEGY EGFP RYPDHMKQHDFFKSAMPEGY c g c t a c c c c g a c c a c a t g a a g c a g c a c g a c t t c t t c a a g t c c g c c a t g c c c g a a g g c t a c g c g a t g g g g c t g g t g t a c t t c g t c g t g c t g a a g a a g t t c a g g c g g t a c g g g c t t c c g a t g

5940 95 100 105 110 VQERTIFFKDDGNYKTRAEV EGFP 95 100 105 110 VQERTIFFKDDGNYKTRAEV EGFP C T G C T G C C G T T G A T G T T C T G EXFP-R VQERTIFFKDDGNYKTRAEV g t c c a g g a g c g c a c c a t c t t c t t c a a g g a c g a c g g c a a c t a c a a g a c c c g c g c c g a g g t g c a g g t c c t c g c g t g g t a g a a g a a g t t c c t g c t g c c g t t g a t g t t c t g g g c g c g g c t c c a c

6000 115 120 125 130 KFEGDTLVNRIELKGIDFKE EGFP 115 120 125 130 KFEGDTLVNRIELKGIDFKE EGFP KFEGDTLVNRIELKGIDFKE a a g t t c g a g g g c g a c a c c c t g g t g a a c c g c a t c g a g c t g a a g g g c a t c g a c t t c a a g g a g t t c a a g c t c c c g c t g t g g g a c c a c t t g g c g t a g c t c g a c t t c c c g t a g c t g a a g t t c c t c

6060 135 140 145 150 DGNILGHKLEYNYNSHNVYI EGFP 135 140 145 150 DGNILGHKLEYNYNSHNVYI EGFP DGNILGHKLEYNYNSHNVYI g a c g g c a a c a t c c t g g g g c a c a a g c t g g a g t a c a a c t a c a a c a g c c a c a a c g t c t a t a t c c t g c c g t t g t a g g a c c c c g t g t t c g a c c t c a t g t t g a t g t t g t c g g t g t t g c a g a t a t a g

6120 155 160 165 170 MADKQKNGIKVNFKIRHNIE EGFP 155 160 165 170 MADKQKNGIKVNFKIRHNIE EGFP MADKQKNGIKVNFKIRHNIE a t g g c c g a c a a g c a g a a g a a c g g c a t c a a g g t g a a c t t c a a g a t c c g c c a c a a c a t c g a g t a c c g g c t g t t c g t c t t c t t g c c g t a g t t c c a c t t g a a g t t c t a g g c g g t g t t g t a g c t c

6180 175 180 185 190 DGSVQLADHYQQNTPIGDGP EGFP 175 180 185 190 DGSVQLADHYQQNTPIGDGP EGFP DGSVQLADHYQQNTPIGDGP g a c g g c a g c g t g c a g c t c g c c g a c c a c t a c c a g c a g a a c a c c c c c a t c g g c g a c g g c c c c c t g c c g t c g c a c g t c g a g c g g c t g g t g a t g g t c g t c t t g t g g g g g t a g c c g c t g c c g g g g

6240 195 200 205 210 VLLPDNHYLSTQSALSKDPN EGFP 195 200 205 210 VLLPDNHYLSTQSALSKDPN EGFP VLLPDNHYLSTQSALSKDPN g t g c t g c t g c c c g a c a a c c a c t a c c t g a g c a c c c a g t c c g c c c t g a g c a a a g a c c c c a a c c a c g a c g a c g g g c t g t t g g t g a t g g a c t c g t g g g t c a g g c g g g a c t c g t t t c t g g g g t t g

6300 215 220 225 230 EKRDHMVLLEFVTAAGITLG EGFP 215 220 225 230 EKRDHMVLLEFVTAAGITLG EGFP C A T G G T C C T G C T G G A G T T C G T G EGFP-C EKRDHMVLLEFVTAAGITLG g a g a a g c g c g a t c a c a t g g t c c t g c t g g a g t t c g t g a c c g c c g c c g g g a t c a c t c t c g g c c t c t t c g c g c t a g t g t a c c a g g a c g a c c t c a a g c a c t g g c g g c g g c c c t a g t g a g a g c c g

BsrGI 6360 235 MDELYK EF* EGFP bGH poly(A) signal 235 MDELYK EF* EGFP G G A G C T G A C A C G G A A BGH-rev MDELYKEF* a t g g a c g a g c t g t a c a a g g a a t t c t a a 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 t a c c t g c t c g a c a t g t t c c t t a a g a t t 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

6420 bGH poly(A) signal G A T BGH-rev c t a g t t g c c a g c c a t c t g t 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 g a t c a a c g g t c g g t a g a c a 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

6480 bGH poly(A) signal c c a c t c c c a c t g t c c t t t c 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 g t g a g g g t g a c a g g a a a 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

6540 bGH poly(A) signal g t c a t t c t a t t c t g g g g g g 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 c a g t a a g a t a a g a c c c c c c 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

NotI 6600 bGH poly(A) signal AAV2 ITR AAV2 ITR a t a g c a g g c a t g c t g g g g a 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 t a t c g t c c g t a c g a c c c 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

6660 AAV2 ITR AAV2 ITR c t c t g c g c g c t c g c t c g c t 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 g a g a c g c g c g a g c g a g c g a 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

SbfIKasINarISfoIPluTI 6720 AAV2 ITR AAV2 ITR t t g c c c g g g c g g c c t c a g t 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 a a c g g g c c c g c c g g a g t 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

6780 g g t a t t t t c t c c t t a c g c a 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 c c a t a a a a g a g g a a t g c g t 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

6840 f1 ori g t a c g c g c c c t g t a g c g g c 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 a t g c g c g g g a c a t c g c c g 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

6900 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 g c t a c a c t t g c c a g c g c c t 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 g c g a t g t g a a c g g t c g c g g a 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

6960 f1 ori c a c g t t c g c c g g c t t t c c c 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 g t g c a a g c g g c c g a a a g g g 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

7020 f1 ori t a g t g c t t t a c g g c a c c t c 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 a t c a c g a a a t g c c g t g g a 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

7080 f1 ori G F1ori-F g c c a t c g c c c t g a t a g a c g 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 c g g t a g c g g g a c t a t c t g c 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

7140 f1 ori T G G A C T C T T G T T C C A A A C T G G F1ori-F t g g a c t c t t g t t c c a a a c t g g a a c a a c a c t c a a c t 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 c c t g a g a a c a a g g t t t g a c c t t g t t g t g a g t t g a 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

PsiI 7200 f1 ori a t a a g g g a t t t t g c c g a t t t c g g t 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 t t c c c t a a a a c g g c t a a a g c c a 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

7260 f1 ori A T G T T A pRS-marker t a a c g c g a a t t t t a a c a a a 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 a t t g c g c t t a a a a t t g t t t 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

7320 G A C G A G A C T A C G G C pRS-marker c t g c t c t g a t g c c g c a t a g 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 g a c g a g a c t a c g g c g t a 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

7380 C C G G G A G pGEX 3' c t g a c g g g c t t g t c t g c t c 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 g a c t g c c c g a a c a g a c g a g 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

7440 C T G C A T G T G T C A G A G G pGEX 3' C G G A G C A pBRforEco c t g c a t g t g t c a g a g g t t t 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 c g t a c a c a g t c t c c a a a 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

7500 C T A T G C G G A T A A pBRforEco g a t a c g c c t a t t t t t a t a g 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 t a t g c g g a t a a a a a t a t c 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

7560 AmpR promoter c a c t t t t c g g g g a a a t g t g 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 g t g a a a a g c c c c t t t a c a c 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

7620 AmpR promoter t a t g t a t c c g c t c a t g a g a 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 a t a c a t a g g c g a g t a c t c 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

7680 1 5 10 15 signal sequence MSIQHFRVALIPFFAAFCL AmpR promoter AmpR MSIQHFRVALIPFFAAFCL g a g t a t g a g t a t t c a a c a t 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 c t c a t a c t c a t a a g t t g t 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

7740 20 signal sequence PVFA 25 30 35 HPETLVKVKDAEDQLG AmpR PVFAHPETLVKVKDAEDQLG t c c t g t t t t t g c t c a c c c a 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 a g g a c a a a a a c g a g t g g g t 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

7800 40 45 50 55 ARVGYIELDLNSGKILESFR AmpR ARVGYIELDLNSGKILESFR t g c a c g a g t g g g t t a c a t c 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 a c g t g c t c a c c c a a t g t a g 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

7860 60 65 70 75 PEERFPMMSTFKVLLCGAVL AmpR C G A T A C A C C G C G C C A T A A Amp-R PEERFPMMSTFKVLLCGAVL c c c c g a a g a a c g t t t t c c a 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 g g g g c t t c t t g c a a a a g g t 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

7920 80 85 90 95 SRIDAGQEQLGRRIHYSQND AmpR T A Amp-R SRIDAGQEQLGRRIHYSQND a t c c c g t a t t g a c g c c g g g 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 t a g g g c a t a a c t g c g g c c c 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

7980 100 105 110 115 LVEYSPVTEKHLTDGMTVRE AmpR LVEYSPVTEKHLTDGMTVRE c t t g g t t g a g t a c t c a c c a 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 g a a c c a a c t c a t g a g t g g t 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

8040 120 125 130 135 LCSAAITMSDNTAANLLLTT AmpR LCSAAITMSDNTAANLLLTT a t t a t g c a g t g c t g c c a t a 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 t a a t a c g t c a c g a c g g t a t 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

PvuI 8100 140 145 150 155 IGGPKELTAFLHNMGDHVTR AmpR IGGPKELTAFLHNMGDHVTR g a t c g g a g g a c c g a a g g a g 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 t a g c c t c c t g g c t t c c t c 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

8160 160 165 170 175 LDRWEPELNEAIPNDERDTT AmpR LDRWEPELNEAIPNDERDTT c c t t g a t c g t t g g g a a c c g 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 g a a c t a g c a a c c c t t g g c 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

FspI 8220 180 185 190 195 MPVAMATTLRKLLTGELLTL AmpR MPVAMATTLRKLLTGELLTL g a t g c c t g t a g c a a t g g c a 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 c t a c g g a c a t c g t t a c c g t 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

8280 200 205 210 215 ASRQQLIDWMEADKVAGPLL AmpR ASRQQLIDWMEADKVAGPLL a g c t t c c c g g c a a c a a t t a 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 t c g a a g g g c c g t t g t t a a 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

8340 220 225 230 235 RSALPAGWFIADKSGAGERG AmpR RSALPAGWFIADKSGAGERG g c g c t c g g c c c t t c c g g c t 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 c g c g a g c c g g g a a g g c c g 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

SacII 8400 240 245 250 255 SRGIIAALGPDGKPSRIVVI AmpR SRGIIAALGPDGKPSRIVVI a a g c c g c g g t a t c a t t g c a 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 t t c g g c g c c a t a g t a a c g t 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

8460 260 265 270 275 YTTGSQATMDERNRQIAEIG AmpR YTTGSQATMDERNRQIAEIG c t a c a c g a c g g g g a g t c a g 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 g a t g t g c t g c c c c t c a g t c 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

8520 280 285 ASLIKHW* AmpR ASLIKHW* t g c c t c a c t g a t t a a g c a t 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 a c g g a g t g a c t a a t t c g t a 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

8580 t g a t t t a a a a c t t c a t t t t 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 a c t a a a t t t t g a a g t a a 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

8640 c a t g a c c a a a a t c c c t t a a 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 t a c t g g t t t t a g g g a a t t 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

8700 ori g a t c a a a g g a t c t t c t t g a 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 c t a g t t t c c t a g a a g a a c t 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

8760 ori a a a a c c a c c g c t a c c a g c g 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 t t t t g g t g g c g a t g g t c 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

8820 ori g a a g g t a a c t g g c t t c a g c a g a g c g c a g a t a c c a a a t a c t g t t c t t c t a g t g t a g c c g t a c t t c c a t t g a c c g a a g t c g t c t c g c g t c t a t g g t t t a t g a c a a g a a g a t c a c a t c g g c a t

8880 ori g t t a g g c c a c c a c t t c a a g 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 c a a t c c g g t g g t g a a g t t 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

8940 ori g t t a c c a g t g g c t g c t g c c 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 c a a t g g t c a c c g a c g a c g 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

9000 ori a t a g t t a c c g g a t a a g g c g 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 t a t c a a t g g c c t a t t c c g 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

9060 ori c t t g g a g c g a a c g a c c t a c 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 g a a c c t c g c t t g c t g g a t g 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

9120 ori c a c g c t t c c c g a a g g g a g a 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 g t g c g a a g g g c t t c c c t c 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

9180 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F a g a g c g c a c g a g g g a g c t t 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 t c g c g t g c t c c c t c g a a 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

9240 ori t c g c c a c c t c t g a c t t g a g 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 a g c g g t g g a g a c t g a a c t c 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

3ʹ 5ʹ 9291 ori g a a a a a c g c c a g c a a c g c g 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 c t t t t t g c g g t c g t t g c g c 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

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.