Addgene PX458_RAD21_1 - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: PX458_RAD21_1


Map View

hU6-F (17 .. 37) AflIII - PciI (9) pBR322ori-F (9148 .. 9167) SacII (8350) FspI (8192) PvuI (8046) Amp-R (7846 .. 7865) pBRforEco (7437 .. 7455) pGEX 3' (7377 .. 7399) pRS-marker (7258 .. 7277) PsiI (7144) F1ori-F (7083 .. 7104) F1ori-R (6873 .. 6892) PluTI (6716) SfoI (6714) NarI (6713) KasI (6712) SbfI (6709) NotI (6564) BGH-rev (6349 .. 6366) BsrGI (6314) EGFP-C (6258 .. 6279) EXFP-R (5911 .. 5930) FseI (5519) BsmI (5027) BsaBI * (4885) LKO.1 5' (188 .. 207) XbaI (447) Acc65I (453) KpnI (457) SnaBI (720) chicken β-actin promoter PaqCI (1236) AgeI (1258) SV40 NLS BglII (1637) PspOMI (2742) ApaI (2746) PasI (3156) EcoRV (3240) PmlI (4176) KflI (4764) PX458_RAD21_1 9294 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 n n 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 a a a a c g a g t g t a c a n n 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

120 U6 promoter c a 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 g t 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

180 U6 promoter a a 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 t 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

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' t a 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 a t 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

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

360 gRNA scaffold 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 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 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 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

420 gRNA scaffold g c 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 c g 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

XbaIAcc65IKpnI 480 CMV enhancer g t 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 c a 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

540 CMV enhancer g g 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 c c 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

600 CMV enhancer g g 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 c 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

660 CMV enhancer c a 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 t 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

SnaBI 720 CMV enhancer g c 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 c g 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

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

840 chicken β- actin promoter a t 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 t a 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

900 chicken β- actin promoter g c 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

960 chicken β- actin promoter c g 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 g c 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

1020 chicken β- actin promoter t t 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 a a 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

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

1140 hybrid intron c c c 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 g g g 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

1200 hybrid intron t c c 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 a g g 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

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

1320 1 5 10 15 DYKDHDGDYKDHDIDY 3xFLAG Kozak sequence MDYKDHDGDYKDHDIDY TGGHVILVVVSVVLIMINIV g g t 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 c c 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

1380 20 KDDDDK MA 130 PKKKRKV (in frame with SV40 NLS) GIHGV 3xFLAG SV40 NLS KDDDDKMAPKKKRKVGIHGV FVIVILHGWLLLPLDTDVSD a a a 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 t t t 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

1440 5 10 15 DKKYSIGLDIGTNSVGW Cas9 PAA 5 10 15 DKKYSIGLDIGTNSVGW Cas9 PAADKKYSIGLDIGTNSVGW WCGVLLVADAQVDAGVRHAP c c a 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 g t 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

1500 20 25 30 35 AVITDEYKVPSKKFKVLGNT Cas9 20 25 30 35 AVITDEYKVPSKKFKVLGNT Cas9 AVITDEYKVPSKKFKVLGNT GHDGVLVLHGALFELHQAVG g c c 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 c g g 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

1560 40 45 50 55 DRHSIKKNLIGALLFDSGET Cas9 40 45 50 55 DRHSIKKNLIGALLFDSGET Cas9 DRHSIKKNLIGALLFDSGET VPVADLLVQDSGQQEVAAFC g a c 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 c t g 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

1620 60 65 70 75 AEATRLKRTARRRYTRRKNR Cas9 60 65 70 75 AEATRLKRTARRRYTRRKNR Cas9 AEATRLKRTARRRYTRRKNR GLGGPQLSGGSSSVGSPLVP g c c 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 c g g 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

BglII 1680 80 85 90 95 ICYLQEIFSNEMAKVDDSFF Cas9 80 85 90 95 ICYLQEIFSNEMAKVDDSFF Cas9 ICYLQEIFSNEMAKVDDSFF DAIQLLDEAVLHGLHVVAEE a t c 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 t a g 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

1740 100 105 110 115 HRLEESFLVEEDKKHERHPI Cas9 100 105 110 115 HRLEESFLVEEDKKHERHPI Cas9 HRLEESFLVEEDKKHERHPI VSQFLGEQHFLILLVLPVGD c a c 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 g t g 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

1800 120 125 130 135 FGNIVDEVAYHEKYPTIYHL Cas9 120 125 130 135 FGNIVDEVAYHEKYPTIYHL Cas9 FGNIVDEVAYHEKYPTIYHL EAVDHVLHGVVLLVGGDVVQ t t c 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 a g 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

1860 140 145 150 155 RKKLVDSTDKADLRLIYLAL Cas9 140 145 150 155 RKKLVDSTDKADLRLIYLAL Cas9 RKKLVDSTDKADLRLIYLAL SLFQHVAGVLGVQPQDIQGQ a g a 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 t c t 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

1920 160 165 170 175 AHMIKFRGHFLIEGDLNPDN Cas9 160 165 170 175 AHMIKFRGHFLIEGDLNPDN Cas9 AHMIKFRGHFLIEGDLNPDN GVHDLEPAVEQDLAVQVGVV g c c 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 c g g 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

1980 180 185 190 195 SDVDKLFIQLVQTYNQLFEE Cas9 180 185 190 195 SDVDKLFIQLVQTYNQLFEE Cas9 SDVDKLFIQLVQTYNQLFEE AVHVLQEDLQHLGVVLQELF a g c 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 t c g 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

2040 200 205 210 215 NPINASGVDAKAILSARLSK Cas9 200 205 210 215 NPINASGVDAKAILSARLSK Cas9 NPINASGVDAKAILSARLSK VGDVGAAHVGLGDQRGSQAL a a c 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 t t g 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

2100 220 225 230 235 SRRLENLIAQLPGEKKNGLF Cas9 220 225 230 235 SRRLENLIAQLPGEKKNGLF Cas9 SRRLENLIAQLPGEKKNGLF ASPQFIQDGLQGALLLIAQE a g c 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 t c g 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

2160 240 245 250 255 GNLIALSLGLTPNFKSNFDL Cas9 240 245 250 255 GNLIALSLGLTPNFKSNFDL Cas9 GNLIALSLGLTPNFKSNFDL SVQNGQAQAQGGVELAVEVQ g g a 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 c c t 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

2220 260 265 270 275 AEDAKLQLSKDTYDDDLDNL Cas9 260 265 270 275 AEDAKLQLSKDTYDDDLDNL Cas9 AEDAKLQLSKDTYDDDLDNL GLIGFQLQALVGVVVVQVVQ g c c 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 g 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

2280 280 285 290 295 LAQIGDQYADLFLAAKNLSD Cas9 280 285 290 295 LAQIGDQYADLFLAAKNLSD Cas9 LAQIGDQYADLFLAAKNLSD QGLDAVLVGVQKQGGLVQGV c t g 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 a 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

2340 300 305 310 315 AILLSDILRVNTEITKAPLS Cas9 300 305 310 315 AILLSDILRVNTEITKAPLS Cas9 AILLSDILRVNTEITKAPLS GDQQAVDQSHVGLDGLGGQA g c c 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 c g g 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

2400 320 325 330 335 ASMIKRYDEHHQDLTLLKAL Cas9 320 325 330 335 ASMIKRYDEHHQDLTLLKAL Cas9 ASMIKRYDEHHQDLTLLKAL GRHDLSVVLVVLVQGQQFSE g c c 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 c g 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

2460 340 345 350 355 VRQQLPEKYKEIFFDQSKNG Cas9 340 345 350 355 VRQQLPEKYKEIFFDQSKNG Cas9 VRQQLPEKYKEIFFDQSKNG HPLLQRLLVFLNEEVLALVA g t g 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 c 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

2520 360 365 370 375 YAGYIDGGASQEEFYKFIKP Cas9 360 365 370 375 YAGYIDGGASQEEFYKFIKP Cas9 YAGYIDGGASQEEFYKFIKP VGAVNVASGALFLEVLEDLG t a c 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 g 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

2580 380 385 390 395 ILEKMDGTEELLVKLNREDL Cas9 380 385 390 395 ILEKMDGTEELLVKLNREDL Cas9 ILEKMDGTEELLVKLNREDL DQFLHVAGLFQEHLQVSLVQ a t c 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 t a 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

2640 400 405 410 415 LRKQRTFDNGSIPHQIHLGE Cas9 400 405 410 415 LRKQRTFDNGSIPHQIHLGE Cas9 LRKQRTFDNGSIPHQIHLGE QPLLPGEVVAADGVLDVQSL c t g 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 g a c 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

2700 420 425 430 435 LHAILRRQEDFYPFLKDNRE Cas9 420 425 430 435 LHAILRRQEDFYPFLKDNRE Cas9 LHAILRRQEDFYPFLKDNRE QVGNQPPLFIKVWEQLVVPF c t g 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 g a c 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

PspOMIApaI 2760 440 445 450 455 KIEKILTFRIPYYVGPLARG Cas9 440 445 450 455 KIEKILTFRIPYYVGPLARG Cas9 KIEKILTFRIPYYVGPLARG LDLLDQGEADGVVHARQGPS a a g 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 t t 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

2820 460 465 470 475 NSRFAWMTRKSEETITPWNF Cas9 460 465 470 475 NSRFAWMTRKSEETITPWNF Cas9 NSRFAWMTRKSEETITPWNF VASEGPHGSLALFGDGGPVE a a c 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 t t 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

2880 480 485 490 495 EEVVDKGASAQSFIERMTNF Cas9 480 485 490 495 EEVVDKGASAQSFIERMTNF Cas9 EEVVDKGASAQSFIERMTNF LFHHVLASGGLAEDLPHGVE g a g 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 c t c 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

2940 500 505 510 515 DKNLPNEKVLPKHSLLYEYF Cas9 500 505 510 515 DKNLPNEKVLPKHSLLYEYF Cas9 DKNLPNEKVLPKHSLLYEYF ILVQGVLLHQGLVAQQVLVE g a t 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 c t a 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

3000 520 525 530 535 TVYNELTKVKYVTEGMRKPA Cas9 520 525 530 535 TVYNELTKVKYVTEGMRKPA Cas9 TVYNELTKVKYVTEGMRKPA GHIVLQGFHFVHGLSHSLGG a c c 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 g g 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

3060 540 545 550 555 FLSGEQKKAIVDLLFKTNRK Cas9 540 545 550 555 FLSGEQKKAIVDLLFKTNRK Cas9 FLSGEQKKAIVDLLFKTNRK EQAALLFLGDHVQQELGVPF t t c 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 a a 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

3120 560 565 570 575 VTVKQLKEDYFKKIECFDSV Cas9 560 565 570 575 VTVKQLKEDYFKKIECFDSV Cas9 VTVKQLKEDYFKKIECFDSV HGHLLQFLVVELFDLAEVGH g t g 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 c a c 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

PasI 3180 580 585 590 595 EISGVEDRFNASLGTYHDLL Cas9 580 585 590 595 EISGVEDRFNASLGTYHDLL Cas9 EISGVEDRFNASLGTYHDLL FDGAHFIPEVGGQACVVIQQ g a a 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 c t t 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

EcoRV 3240 600 605 610 615 KIIKDKDFLDNEENEDILED Cas9 600 605 610 615 KIIKDKDFLDNEENEDILED Cas9 KIIKDKDFLDNEENEDILED FNDLVLVEQVILFVLVNQFI a a a 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 t t t 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

3300 620 625 630 635 IVLTLTLFEDREMIEERLKT Cas9 620 625 630 635 IVLTLTLFEDREMIEERLKT Cas9 IVLTLTLFEDREMIEERLKT DHQGQCQKLVSLHDLFPQFG a t c 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 g 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

3360 640 645 650 655 YAHLFDDKVMKQLKRRRYTG Cas9 640 645 650 655 YAHLFDDKVMKQLKRRRYTG Cas9 YAHLFDDKVMKQLKRRRYTG *AWRNSSLTIFCSFRRLYVP IGVQEVVFHHLLQLPPSVGA t a t 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 a t a 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

3420 660 665 670 675 WGRLSRKLINGIRDKQSGKT Cas9 660 665 670 675 WGRLSRKLINGIRDKQSGKT Cas9 WGRLSRKLINGIRDKQSGKT QPLSLRFSILPMRSLCDPLV PAPQAPLQDVADPVLLGALC t g g 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 c 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

3480 680 685 690 695 ILDFLKSDGFANRNFMQLIH Cas9 680 685 690 695 ILDFLKSDGFANRNFMQLIH Cas9 ILDFLKSDGFANRNFMQLIH IRSKRFDSPKALLFKMCSIW DQIEQLGVAEGVSVEHLQDV a t c 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 t a g 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

3540 700 705 710 715 DDSLTFKEDIQKAQVSGQGD Cas9 700 705 710 715 DDSLTFKEDIQKAQVSGQGD Cas9 DDSLTFKEDIQKAQVSGQGD SSLRVKLSSMWFAWTDPWPS VVAQGKFLVDLFGLHGALAI g a c 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 c t g 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

3600 720 725 730 735 SLHEHIANLAGSPAIKKGIL Cas9 720 725 730 735 SLHEHIANLAGSPAIKKGIL Cas9 SLHEHIANLAGSPAIKKGIL LRCSCMALRAPLGAMLFPMR AQVLVNGIQGAAGGNLLADQ a g c 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 t c 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

3660 740 745 750 755 QTVKVVDELVKVMGRHKPEN Cas9 740 745 750 755 QTVKVVDELVKVMGRHKPEN Cas9 QTVKVVDELVKVMGRHKPEN CVTFTTSSSTFTIPRCLGSF LCHLHHVLEHFHHAPVLGLV c a g 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 g 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

3720 760 765 770 775 IVIEMARENQTTQKGQKNSR Cas9 760 765 770 775 IVIEMARENQTTQKGQKNSR Cas9 IVIEMARENQTTQKGQKNSR MTISIALSFWVVWFPCFFLR DHDFHGSLVLGGLLSLLVAA a t c 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 t 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

3780 780 785 790 795 ERMKRIEEGIKELGSQILKE Cas9 780 785 790 795 ERMKRIEEGIKELGSQILKE Cas9 ERMKRIEEGIKELGSQILKE SLIFRISSPM LSHLPDFLADFLQAALDQFF g a g 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 t 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

3840 800 805 810 815 HPVENTQLQNEKLYLYYLQN Cas9 800 805 810 815 HPVENTQLQNEKLYLYYLQN Cas9 HPVENTQLQNEKLYLYYLQN VGHFVGLQLVLLQVQVVQLI c a c 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 t 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

3900 820 825 830 835 GRDMYVDQELDINRLSDYDV Cas9 820 825 830 835 GRDMYVDQELDINRLSDYDV Cas9 GRDMYVDQELDINRLSDYDV PPIHVHVLFQVDVPQGVVIH g g g 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 c c 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

3960 840 845 850 855 DHIVPQSFLKDDSIDNKVLT Cas9 840 845 850 855 DHIVPQSFLKDDSIDNKVLT Cas9 DHIVPQSFLKDDSIDNKVLT VM g a c 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 c t g 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

4020 860 865 870 875 RSDKNRGKSDNVPSEEVVKK Cas9 860 865 870 875 RSDKNRGKSDNVPSEEVVKK Cas9 RSDKNRGKSDNVPSEEVVKK a g a 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 t c t 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

4080 880 885 890 895 MKNYWRQLLNAKLITQRKFD Cas9 880 885 890 895 MKNYWRQLLNAKLITQRKFD Cas9 MKNYWRQLLNAKLITQRKFD a t g 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 t a c 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

4140 900 905 910 915 NLTKAERGGLSELDKAGFIK Cas9 900 905 910 915 NLTKAERGGLSELDKAGFIK Cas9 NLTKAERGGLSELDKAGFIK a a t 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 t t 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

PmlI 4200 920 925 930 935 RQLVETRQITKHVAQILDSR Cas9 920 925 930 935 RQLVETRQITKHVAQILDSR Cas9 RQLVETRQITKHVAQILDSR a g a 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 t c t 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

4260 940 945 950 955 MNTKYDENDKLIREVKVITL Cas9 940 945 950 955 MNTKYDENDKLIREVKVITL Cas9 MNTKYDENDKLIREVKVITL a t g 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 t a c 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

4320 960 965 970 975 KSKLVSDFRKDFQFYKVREI Cas9 960 965 970 975 KSKLVSDFRKDFQFYKVREI Cas9 KSKLVSDFRKDFQFYKVREI a a g 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 t t 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

4380 980 985 990 995 NNYHHAHDAYLNAVVGTALI Cas9 980 985 990 995 NNYHHAHDAYLNAVVGTALI Cas9 NNYHHAHDAYLNAVVGTALI a a c 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 t t g 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

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

4500 1020 1025 1030 1035 RKMIAKSEQEIGKATAKYFF Cas9 1020 1025 1030 1035 RKMIAKSEQEIGKATAKYFF Cas9 RKMIAKSEQEIGKATAKYFF c g g 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 g c 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

4560 1040 1045 1050 1055 YSNIMNFFKTEITLANGEIR Cas9 1040 1045 1050 1055 YSNIMNFFKTEITLANGEIR Cas9 YSNIMNFFKTEITLANGEIR t a c 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 t 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

4620 1060 1065 1070 1075 KRPLIETNGETGEIVWDKGR Cas9 1060 1065 1070 1075 KRPLIETNGETGEIVWDKGR Cas9 KRPLIETNGETGEIVWDKGR a a g 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 t t c 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

4680 1080 1085 1090 1095 DFATVRKVLSMPQVNIVKKT Cas9 1080 1085 1090 1095 DFATVRKVLSMPQVNIVKKT Cas9 DFATVRKVLSMPQVNIVKKT g a t 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 c t a 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

4740 1100 1105 1110 1115 EVQTGGFSKESILPKRNSDK Cas9 1100 1105 1110 1115 EVQTGGFSKESILPKRNSDK Cas9 EVQTGGFSKESILPKRNSDK g a g 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 c 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

KflI 4800 1120 1125 1130 1135 LIARKKDWDPKKYGGFDSPT Cas9 1120 1125 1130 1135 LIARKKDWDPKKYGGFDSPT Cas9 LIARKKDWDPKKYGGFDSPT c t g 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 a c 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

4860 1140 1145 1150 1155 VAYSVLVVAKVEKGKSKKLK Cas9 1140 1145 1150 1155 VAYSVLVVAKVEKGKSKKLK Cas9 VAYSVLVVAKVEKGKSKKLK g t g 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 c a c 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

BsaBI* 4920 1160 1165 1170 1175 SVKELLGITIMERSSFEKNP Cas9 1160 1165 1170 1175 SVKELLGITIMERSSFEKNP Cas9 SVKELLGITIMERSSFEKNP a g t 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 t c a 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

4980 1180 1185 1190 1195 IDFLEAKGYKEVKKDLIIKL Cas9 1180 1185 1190 1195 IDFLEAKGYKEVKKDLIIKL Cas9 IDFLEAKGYKEVKKDLIIKL a t c 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 t a g 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

BsmI 5040 1200 1205 1210 1215 PKYSLFELENGRKRMLASAG Cas9 1200 1205 1210 1215 PKYSLFELENGRKRMLASAG Cas9 PKYSLFELENGRKRMLASAG c c t 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 g 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

5100 1220 1225 1230 1235 ELQKGNELALPSKYVNFLYL Cas9 1220 1225 1230 1235 ELQKGNELALPSKYVNFLYL Cas9 ELQKGNELALPSKYVNFLYL g a a 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 c t t 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

5160 1240 1245 1250 1255 ASHYEKLKGSPEDNEQKQLF Cas9 1240 1245 1250 1255 ASHYEKLKGSPEDNEQKQLF Cas9 ASHYEKLKGSPEDNEQKQLF g c c 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 c g 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

5220 1260 1265 1270 1275 VEQHKHYLDEIIEQISEFSK Cas9 1260 1265 1270 1275 VEQHKHYLDEIIEQISEFSK Cas9 VEQHKHYLDEIIEQISEFSK g t g 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 c a c 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

5280 1280 1285 1290 1295 RVILADANLDKVLSAYNKHR Cas9 1280 1285 1290 1295 RVILADANLDKVLSAYNKHR Cas9 RVILADANLDKVLSAYNKHR a g a 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 t c 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

5340 1300 1305 1310 1315 DKPIREQAENIIHLFTLTNL Cas9 1300 1305 1310 1315 DKPIREQAENIIHLFTLTNL Cas9 DKPIREQAENIIHLFTLTNL g a t 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 c t 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

5400 1320 1325 1330 1335 GAPAAFKYFDTTIDRKRYTS Cas9 1320 1325 1330 1335 GAPAAFKYFDTTIDRKRYTS Cas9 GAPAAFKYFDTTIDRKRYTS g g a 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 c c t 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

5460 1340 1345 1350 1355 TKEVLDATLIHQSITGLYET Cas9 1340 1345 1350 1355 TKEVLDATLIHQSITGLYET Cas9 TKEVLDATLIHQSITGLYET a c c 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 t 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

FseI 5520 1360 1365 RIDLSQLGGD 1 5 10 KRPAATKKAG Cas9 nucleoplasmin NLS 1360 1365 RIDLSQLGGD (in frame with Cas9) KRPAATKKAG Cas9 RIDLSQLGGDKRPAATKKAG c g g 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 g c 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 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

5580 (in frame with Cas9) QAKKKKEFGSG 140 145 EGRGSLLTC T2A 15 QAKKKK (in frame with nucleoplasmin NLS) EFGSGEGRGSLLTC nucleoplasmin NLS QAKKKKEFGSGEGRGSLLTC c a g 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 t c 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

5640 (in frame with nucleoplasmin NLS) GDVEENPGP 1a V 5 10 SKGEELFTGV EGFP 150 155 GDVEENPGP 1a V 5 10 SKGEELFTGV T2A EGFP GDVEENPGPVSKGEELFTGV g g t 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 c c a 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

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

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

5820 55 60 65 70 KLPVPWPTLVTTLTYGVQCF EGFP 55 60 65 70 KLPVPWPTLVTTLTYGVQCF EGFP KLPVPWPTLVTTLTYGVQCF a a g 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 t t 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

5880 75 80 85 90 SRYPDHMKQHDFFKSAMPEG EGFP 75 80 85 90 SRYPDHMKQHDFFKSAMPEG EGFP SRYPDHMKQHDFFKSAMPEG a g c 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 c g 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

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

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

6060 135 140 145 150 EDGNILGHKLEYNYNSHNVY EGFP 135 140 145 150 EDGNILGHKLEYNYNSHNVY EGFP EDGNILGHKLEYNYNSHNVY g a g 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 c 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

6120 155 160 165 170 IMADKQKNGIKVNFKIRHNI EGFP 155 160 165 170 IMADKQKNGIKVNFKIRHNI EGFP IMADKQKNGIKVNFKIRHNI a t c 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 t 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

6180 175 180 185 190 EDGSVQLADHYQQNTPIGDG EGFP 175 180 185 190 EDGSVQLADHYQQNTPIGDG EGFP EDGSVQLADHYQQNTPIGDG g a g 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 t 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

6240 195 200 205 210 PVLLPDNHYLSTQSALSKDP EGFP 195 200 205 210 PVLLPDNHYLSTQSALSKDP EGFP PVLLPDNHYLSTQSALSKDP c c c 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 g g g 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

6300 215 220 225 230 NEKRDHMVLLEFVTAAGITL EGFP 215 220 225 230 NEKRDHMVLLEFVTAAGITL 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 NEKRDHMVLLEFVTAAGITL a a c 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 t t g 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

BsrGI 6360 235 GMDELYK EF* EGFP bGH poly(A) signal 235 GMDELYK EF* EGFP G G A G C T G A C A C G BGH-rev GMDELYKEF* g g c 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 c g 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

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

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

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

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

6660 AAV2 ITR AAV2 ITR c c t 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 g a 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

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

6780 t g c 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 c g 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

6840 f1 ori a t a 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 t a t 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

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

6960 f1 ori c g c 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 g c g 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

7020 f1 ori a t t 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 a a 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

7080 f1 ori t g g 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 a c c 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

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

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

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

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

7380 C C G G pGEX 3' g c c 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 c g 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

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

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

7560 AmpR promoter t g g 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 c c 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

7620 AmpR promoter a a a 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 t t t 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

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

7740 20 signal sequence LPVFA 25 30 35 HPETLVKVKDAEDQL AmpR LPVFAHPETLVKVKDAEDQL c c t 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 a 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

7800 40 45 50 55 GARVGYIELDLNSGKILESF AmpR GARVGYIELDLNSGKILESF g g g 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 c c c 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

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

7920 80 85 90 95 LSRIDAGQEQLGRRIHYSQN AmpR T A A T A Amp-R LSRIDAGQEQLGRRIHYSQN a t t 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 a 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

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

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

PvuI 8100 140 145 150 155 TIGGPKELTAFLHNMGDHVT AmpR TIGGPKELTAFLHNMGDHVT a a c 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 t 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

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

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

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

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

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

8460 260 265 270 275 IYTTGSQATMDERNRQIAEI AmpR IYTTGSQATMDERNRQIAEI t a t 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 t a 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

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

8580 g a t 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 c t a 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

8640 t c t 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 g 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

8700 ori a a a 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 t t t 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

8760 ori a a a 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 t t 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

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

8880 ori g t a 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 a 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

8940 ori c c t 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 g g a 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

9000 ori a c g 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 t g c 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

9060 ori c a g 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 g t 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

9120 ori c g c 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 g c 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

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 g 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 t c c 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

9240 ori g t t 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 c a a 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

3ʹ 5ʹ 9294 ori a t g 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 t a 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.