Addgene lentiCRISPRv2-7SK-Blast Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: lentiCRISPRv2-7SK-Blast Sequencing Result


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Acc65I (12,805) PacI (12,801) gp41 peptide AleI (12,157) BbvCI (12,004) NotI (11,728) PluTI (11,225) SfoI (11,223) NarI (11,222) KasI (11,221) CMV-F (10,975 .. 10,995) SnaBI (10,796) NdeI (10,690) SpeI (10,455) pRS-marker (10,246 .. 10,265) SgrDI (10,200) AmpR promoter SspI (10,083) Amp-R (9829 .. 9848) FspI (9501) pBR322ori-F (8527 .. 8546) PciI (8386) L4440 (8276 .. 8293) CAP binding site lac promoter M13/pUC Reverse (8058 .. 8080) lac operator M13 Reverse (8045 .. 8061) M13 rev BstZ17I (8007) EBV-rev (7966 .. 7985) SV40pA-R (7912 .. 7931) FseI (7647) SgrAI (7485) EM7 promoter AvrII (7242) StuI (7241) SV40pro-F (7170 .. 7189) pBABE 3' (6923 .. 6943) F1ori-F (6783 .. 6804) F1ori-R (6573 .. 6592) KpnI (12,809) HpaI (253) EcoRI (347) NheI (353) BmtI (357) AgeI (615) XbaI (621) AfeI (628) PasI (2407) EcoRV (2491) nucleoplasmin NLS FLAG BamHI (4814) MluI (5276) WPRE-R (5344 .. 5364) SacII (5794) PmeI (6194) BGH-rev (6210 .. 6227) lentiCRISPRv2-7SK-Blast 12,810 bp

Sequence View

5ʹ 3ʹ 60 c t g c a g t a t t t a g c a t g c c c c a c c c a t c t g c a a g g c a t t c t g g a t a g t g t c a a a a c a g c c g a c g t c a t a a a t c g t a c g g g g t g g g t a g a c g t t c c g t a a g a c c t a t c a c a g t t t t g t c g g

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

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

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

HpaI 300 gRNA scaffold * a c c g g a g a c g g t t a a c a c g t c t c 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 t g g c c t c t g c c a a t t g t g c a g a g 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

BmtINheIEcoRI 360 gRNA scaffold DTILKFFHCRTPAKKFESAL a g t c c g t t a t c a a c t t g a a a a a g t g g c a c c g a g t c g g t g c t t t t t t g a a t t c g c t a g c t a t c a g g c a a t a g t t g a a c t t t t t c a c c g t g g c t c a g c c a c g a a a a a a c t t a a g c g a t c g a t

420 EF-1 α core promoter DQFSHSNAGTGTLPCLACRG g g t c t t g a a a g g a g t g g g a a t t g g c t c c g g t g c c c g t c a g t g g g c a g a g c g c a c a t c g c c c c a g a a c t t t c c t c a c c c t t a a c c g a g g c c a c g g g c a g t c a c c c g t c t c g c g t g t a g c g g

480 EF-1 α core promoter CDGLLQPSPDAISGTGLSPP c a c a g t c c c c g a g a a g t t g g g g g g a g g g g t c g g c a a t t g a t c c g g t g c c t a g a g a a g g t g g t g t c a g g g g c t c t t c a a c c c c c c t c c c c a g c c g t t a a c t a g g c c a c g g a t c t c t t c c a c

540 EF-1 α core promoter APYVPFTIDHVPEAKKGLTP g c g c g g g g t a a a c t g g g a a a g t g a t g t c g t g t a c t g g c t c c g c c t t t t t c c c g a g g g t g g c g c g c c c c a t t t g a c c c t t t c a c t a c a g c a c a t g a c c g a g g c g g a a a a a g g g c t c c c a c c

600 EF-1 α core promoter SFRIYTCYDGHVNKKAVPKG g g g a g a a c c g t a t a t a a g t g c a g t a g t c g c c g t g a a c g t t c t t t t t c g c a a c g g g t t t g c c c c t c t t g g c a t a t a t t c a c g t c a t c a g c g g c a c t t g c a a g a a a a a g c g t t g c c c a a a c g

AfeIXbaIAgeI 660 1 5 MDKKYSIG EF-1 α core promoter Cas9 1 5 MDKKYSIG Cas9 Kozak sequence MDKKYSIG GSCLVPELASGGHVLLVADA c g c c a g a a c a c a g g a c c g g t t c t a g a g c g c t g c c a c c a t g g a c a a g a a g t a c a g c a t c g g g c g g t c t t g t g t c c t g g c c a a g a t c t c g c g a c g g t g g t a c c t g t t c t t c a t g t c g t a g c c

720 10 15 20 25 LDIGTNSVGWAVITDEYKVP Cas9 10 15 20 25 LDIGTNSVGWAVITDEYKVP Cas9 LDIGTNSVGWAVITDEYKVP QVDAGVRHAPGHDGVLVLHG 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 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 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 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

780 30 35 40 45 SKKFKVLGNTDRHSIKKNLI Cas9 30 35 40 45 SKKFKVLGNTDRHSIKKNLI Cas9 SKKFKVLGNTDRHSIKKNLI ALFELHQAVGVPVADLLVQD 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 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 g t c g t t c t t t a a g t t c c a c g a c c c g t t g t g g c t g g 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

840 50 55 60 65 GALLFDSGETAEATRLKRTA Cas9 50 55 60 65 GALLFDSGETAEATRLKRTA Cas9 GALLFDSGETAEATRLKRTA SGQQEVAAFCGLGGPQLSGG 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 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 g c c t c g g g a c g a c a a g c t g t c g c c g c t t t g t c g g c 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

900 70 75 80 85 RRRYTRRKNRICYLQEIFSN Cas9 70 75 80 85 RRRYTRRKNRICYLQEIFSN Cas9 RRRYTRRKNRICYLQEIFSN SSSVGSPLVPDAIQLLDEAV c a g a a g a a g a t a c a c c a g a c g g a a g a a c c g g a t c t 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 g t c t t c t t c t a t g t g g t c t g c c t t c t t g g c c t a g a 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

960 90 95 100 105 EMAKVDDSFFHRLEESFLVE Cas9 90 95 100 105 EMAKVDDSFFHRLEESFLVE Cas9 EMAKVDDSFFHRLEESFLVE LHGLHVVAEEVSQFLGEQHF 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 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 g c t c t a c c g g t t c c a c c t g c t g t c g a a g a a g g t g t 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

1020 110 115 120 125 EDKKHERHPIFGNIVDEVAY Cas9 110 115 120 125 EDKKHERHPIFGNIVDEVAY Cas9 EDKKHERHPIFGNIVDEVAY LILLVLPVGDEAVDHVLHGV a g a g g a t a a g a a g c a c g a g c g g c a c c c c a t c t t c g 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 t c t c c t a t t c t t c g t g c t c g c c g t g g g g t a g a a g c 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

1080 130 135 140 145 HEKYPTIYHLRKKLVDSTDK Cas9 130 135 140 145 HEKYPTIYHLRKKLVDSTDK Cas9 HEKYPTIYHLRKKLVDSTDK VLLVGGDVVQSLFQHVAGVL c c a c g a g a a g t a c c c c a c c a t c t a c c a c c t g a g a a 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 t g c t c t t c a t g g g g t g g t a g a t g g t g g a c t c t t 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

1140 150 155 160 165 ADLRLIYLALAHMIKFRGHF Cas9 150 155 160 165 ADLRLIYLALAHMIKFRGHF Cas9 ADLRLIYLALAHMIKFRGHF GVQPQDIQGQGVHDLEPAVE g g c c g a c c t g c g g c t g a t c t a t c t g g c c c t g g c c c 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 g g c t g g a c g c c g a c t a g a t a g a c c g g g a c c g g g 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

1200 170 175 180 185 LIEGDLNPDNSDVDKLFIQL Cas9 170 175 180 185 LIEGDLNPDNSDVDKLFIQL Cas9 LIEGDLNPDNSDVDKLFIQL QDLAVQVGVVAVHVLQEDLQ 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 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 a c t a g c t c c c g c t g g a c t t g g g g c t g t t g t c g c 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

1260 190 195 200 205 VQTYNQLFEENPINASGVDA Cas9 190 195 200 205 VQTYNQLFEENPINASGVDA Cas9 VQTYNQLFEENPINASGVDA HLGVVLQELFVGDVGAAHVG 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 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 c a c g t c t g g a t g t t g g t c g a c a a g c t c c t t t t g g 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

1320 210 215 220 225 KAILSARLSKSRRLENLIAQ Cas9 210 215 220 225 KAILSARLSKSRRLENLIAQ Cas9 KAILSARLSKSRRLENLIAQ LGDQRGSQALASPQFIQDGL c a a g g c c a t c c t g t c t g c c a g a c t g a g c a a g a g c a g 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 t t c c g g t a g g a c a g a c g g t c t g a c t c g t t c t c g t 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

1380 230 235 240 245 LPGEKKNGLFGNLIALSLGL Cas9 230 235 240 245 LPGEKKNGLFGNLIALSLGL Cas9 LPGEKKNGLFGNLIALSLGL QGALLLIAQESVQNGQAQAQ 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 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 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 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

1440 250 255 260 265 TPNFKSNFDLAEDAKLQLSK Cas9 250 255 260 265 TPNFKSNFDLAEDAKLQLSK Cas9 TPNFKSNFDLAEDAKLQLSK GGVELAVEVQGLIGFQLQAL g a c c c c c a a c t t c a a g a g c a a c t t c g a c c t g g c c g 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 c t g g g g g t t g a a g t t c t c g t t g a a g c t g g a c c g g c 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

1500 270 275 280 285 DTYDDDLDNLLAQIGDQYAD Cas9 270 275 280 285 DTYDDDLDNLLAQIGDQYAD Cas9 DTYDDDLDNLLAQIGDQYAD VGVVVVQVVQQGLDAVLVGV g g a c a c c t a c g a c g a c g a c c t g g a c a a c c t g c t g g 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 g g a t g c t g c t g c t g g a c c t g t t g g a c g a c c 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

1560 290 295 300 305 LFLAAKNLSDAILLSDILRV Cas9 290 295 300 305 LFLAAKNLSDAILLSDILRV Cas9 LFLAAKNLSDAILLSDILRV QKQGGLVQGVGDQQAVDQSH c c t g t t t c t g g c c g c c a a g a a c c t g t c c g a c g c c a 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 g a c a a a g a c c g g c g g t t c t t g g a c a g g c t g c g g t 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

1620 310 315 320 325 NTEITKAPLSASMIKRYDEH Cas9 310 315 320 325 NTEITKAPLSASMIKRYDEH Cas9 NTEITKAPLSASMIKRYDEH VGLDGLGGQAGRHDLSVVLV 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 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 t t g t g g c t c t a g t g g t t c c g g g g g g a c t c g c g g a 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

1680 330 335 340 345 HQDLTLLKALVRQQLPEKYK Cas9 330 335 340 345 HQDLTLLKALVRQQLPEKYK Cas9 HQDLTLLKALVRQQLPEKYK VLVQGQQFSEHPLLQRLLVF c c a c c a g g a c c t g a c c c t g c t g a a a g c t c t c g t g c g g c a g c a g c t g c c t g a g a a g t a c a a g g t g g t c c t g g a c t g g g a c g a c t t t c g a g a g c a c g 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

1740 350 355 360 365 EIFFDQSKNGYAGYIDGGAS Cas9 350 355 360 365 EIFFDQSKNGYAGYIDGGAS Cas9 EIFFDQSKNGYAGYIDGGAS LNEEVLALVAVGAVNVASGA 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 g c t a c a t t g a c g g c g g a g c c a g t c t c t a a a a g a a g c t g g t c t c g t t c t t g c c g a t g c 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

1800 370 375 380 385 QEEFYKFIKPILEKMDGTEE Cas9 370 375 380 385 QEEFYKFIKPILEKMDGTEE Cas9 QEEFYKFIKPILEKMDGTEE LFLEVLEDLGDQFLHVAGLF 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 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 g g t c c t t c t c a a g a t g t t c a a g t a g t t c g g g t a g g 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

1860 390 395 400 405 LLVKLNREDLLRKQRTFDNG Cas9 390 395 400 405 LLVKLNREDLLRKQRTFDNG Cas9 LLVKLNREDLLRKQRTFDNG QEHLQVSLVQQPLLPGEVVA a c t g c t c g t g a a g c t g a a c a g a g a g g a c c t g c t g c 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 t g a c g a g c a c t t c g a c t t g t c t c t c c t g g a c g a c g 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

1920 410 415 420 425 SIPHQIHLGELHAILRRQED Cas9 410 415 420 425 SIPHQIHLGELHAILRRQED Cas9 SIPHQIHLGELHAILRRQED ADGVLDVQSLQVGNQPPLFI c a g c a t c c c c c a c c a g a t c c a c c t g g g a g a g c t g c 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 g t c g t a g g g g g t g g t c t a g g t g g a c c c t c t c g a c g 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

1980 430 435 440 445 FYPFLKDNREKIEKILTFRI Cas9 430 435 440 445 FYPFLKDNREKIEKILTFRI Cas9 FYPFLKDNREKIEKILTFRI KVWEQLVVPFLDLLDQGEAD t t t t t a c c c a t t c c t g a a g g a c a a c c g g g a a a a g a t c g a g a a g a t c c t g a c c t t c c g c a t a a a a a t g g g t a a g g a c t t c c t g t t g g c c c t t t t c t 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

2040 450 455 460 465 PYYVGPLARGNSRFAWMTRK Cas9 450 455 460 465 PYYVGPLARGNSRFAWMTRK Cas9 PYYVGPLARGNSRFAWMTRK GVVHARQGPSVASEGPHGSL c c c c t a c t a c g t g g g c c c t c t g g c c a g g g g a a a c a 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 g g g a t g a t g c a c c c g g g a g a c c g g t c c c c t t t g t 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

2100 470 475 480 485 SEETITPWNFEEVVDKGASA Cas9 470 475 480 485 SEETITPWNFEEVVDKGASA Cas9 SEETITPWNFEEVVDKGASA ALFGDGGPVELFHHVLASGG g a g c g a g g a a a c c a t c a c c c c c t g g a a c t t c g a g g 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 t c g c t c c t t t g g t a g t g g g g g a c c t t g a a g c t c c 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

2160 490 495 500 505 QSFIERMTNFDKNLPNEKVL Cas9 490 495 500 505 QSFIERMTNFDKNLPNEKVL Cas9 QSFIERMTNFDKNLPNEKVL LAEDLPHGVEILVQGVLLHQ c c a g a g c t t c a t c g a g c g g a t g a c c a a c t t c g a t a 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 g t c t c g a a g t a g c t c g c c t a c t g g t t g a a g c t a t 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

2220 510 515 520 525 PKHSLLYEYFTVYNELTKVK Cas9 510 515 520 525 PKHSLLYEYFTVYNELTKVK Cas9 PKHSLLYEYFTVYNELTKVK GLVAQQVLVEGHIVLQGFHF g c c c a a g c a c a g c c t g c t g t a c g a g t a c t t c a c c g t g t a t a a c g a g c t g a c c a a a g t g a a c g g g t t c g t g t c g g a c g a c a t g c t c a t g a a g t g g c 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

2280 530 535 540 545 YVTEGMRKPAFLSGEQKKAI Cas9 530 535 540 545 YVTEGMRKPAFLSGEQKKAI Cas9 YVTEGMRKPAFLSGEQKKAI VHGLSHSLGGEQAALLFLGD 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 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 t a t g c a c t g g c t c c c t t a c t c t t t c g g g c g g a a g g 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

2340 550 555 560 565 VDLLFKTNRKVTVKQLKEDY Cas9 550 555 560 565 VDLLFKTNRKVTVKQLKEDY Cas9 VDLLFKTNRKVTVKQLKEDY HVQQELGVPFHGHLLQFLVV c g t g g a c c t g c t g t t c a a g a c c a a c c g g a a a g t g a 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 g c a c c t g g a c g a c a a g t t c t g g t t g g c c t t t c a c t 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

2400 570 575 580 585 FKKIECFDSVEISGVEDRFN Cas9 570 575 580 585 FKKIECFDSVEISGVEDRFN Cas9 FKKIECFDSVEISGVEDRFN ELFDLAEVGHFDGAHFIPEV c t t c a a g a a a a t c g a g t g c t t c g a c t c c g t g g a a a 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 g a a g t t c t t t t a g c t c a c g a a g c t g a g g c a c c t t t 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

PasI 2460 590 595 600 605 ASLGTYHDLLKIIKDKDFLD Cas9 590 595 600 605 ASLGTYHDLLKIIKDKDFLD Cas9 ASLGTYHDLLKIIKDKDFLD GGQACVVIQQFNDLVLVEQV c g c c t c c c t g g g c a c a t a c c a c g a t c t g c t g a a a a 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 g c g g a g g g a c c c g t g t a t g g t g c t a g a c g a c t t t t 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

EcoRV 2520 610 615 620 625 NEENEDILEDIVLTLTLFED Cas9 610 615 620 625 NEENEDILEDIVLTLTLFED Cas9 NEENEDILEDIVLTLTLFED ILFVLVNQFIDHQGQCQKLV c a a t g a g g a a a a c g a g g a c a t t c t g g a a g a t a t c g 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 g t t a c t c c t t t t g c t c c t g t a a g a c c t t c t a t a g c 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

2580 630 635 640 645 REMIEERLKTYAHLFDDKVM Cas9 630 635 640 645 REMIEERLKTYAHLFDDKVM Cas9 REMIEERLKTYAHLFDDKVM SLHDLFPQFGIGVQEVVFHH *AWRNSSLTI c a g a g a g a t g a t c g a g g a a c g g c t g a a a a c c t a t g 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 t c t c t c t a c t a g c t c c t t g c c g a c t t t t g g a t a c 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

2640 650 655 660 665 KQLKRRRYTGWGRLSRKLIN Cas9 650 655 660 665 KQLKRRRYTGWGRLSRKLIN Cas9 KQLKRRRYTGWGRLSRKLIN LLQLPPSVGAPAPQAPLQDV FCSFRRLYVPQPLSLRFSIL g a a g c a g c t g a a g c g g c g g a g a t a c a c c g g c t g g g g 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 t t c g t c g a c t t c g c c g c c t c t a t g t g g c c g a c c c 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

2700 670 675 680 685 GIRDKQSGKTILDFLKSDGF Cas9 670 675 680 685 GIRDKQSGKTILDFLKSDGF Cas9 GIRDKQSGKTILDFLKSDGF ADPVLLGALCDQIEQLGVAE PMRSLCDPLVIRSKRFDSPK 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 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 g c c g t a g g c c c t g t t c g t c a g g c c g t t c t g t t a g g 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

2760 690 695 700 705 ANRNFMQLIHDDSLTFKEDI Cas9 690 695 700 705 ANRNFMQLIHDDSLTFKEDI Cas9 ANRNFMQLIHDDSLTFKEDI GVSVEHLQDVVVAQGKFLVD ALLFKMCSIWSSLRVKLSSM 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 a g c c t g a c c t t t a a a g a g g a c a t g c g g t t g t c t t t g a a g t a c g t c g a c t a g g t g c t g c 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

2820 710 715 720 725 QKAQVSGQGDSLHEHIANLA Cas9 710 715 720 725 QKAQVSGQGDSLHEHIANLA Cas9 QKAQVSGQGDSLHEHIANLA LFGLHGALAIAQVLVNGIQG WFAWTDPWPSLRCSCMALRA 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 c a c g a g c a c a t t g c c a a t c t g g c g g t c t t t c g g g t c c a c a g g c c g g t c c c g c t a t c g g 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

2880 730 735 740 745 GSPAIKKGILQTVKVVDELV Cas9 730 735 740 745 GSPAIKKGILQTVKVVDELV Cas9 GSPAIKKGILQTVKVVDELV AAGGNLLADQLCHLHHVLEH PLGAMLFPMRCVTFTTSSST c g g c a g c c c c g c c a t t a a g a a g g g c a t c c t g c a g a 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 c c g t c g g g g c g g t a a t t c t t c c c g t a g g a c g t c t 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

2940 750 755 760 765 KVMGRHKPENIVIEMARENQ Cas9 750 755 760 765 KVMGRHKPENIVIEMARENQ Cas9 KVMGRHKPENIVIEMARENQ FHHAPVLGLVDHDFHGSLVL FTIPRCLGSFMTISIALSFW g a a a g t g a t g g g c c g g c a c a a g c c c g a g a a c a t c g 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 c t t t c a c t a c c c g g c c g t g t t c g g g c t c t t g t a g c 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

3000 770 775 780 785 TTQKGQKNSRERMKRIEEGI Cas9 770 775 780 785 TTQKGQKNSRERMKRIEEGI Cas9 TTQKGQKNSRERMKRIEEGI GGLLSLLVAALSHLPDFLAD VVWFPCFFLRSLIFRISSPM g a c c a c c c a g a a g g g a c a g a a g a a c a g c c g c g a g a 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 t g g t g g g t c t t c c c t g t c t t c t t g t c g g c g c t c t 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

3060 790 795 800 805 KELGSQILKEHPVENTQLQN Cas9 790 795 800 805 KELGSQILKEHPVENTQLQN Cas9 KELGSQILKEHPVENTQLQN FLQAALDQFFVGHFVGLQLV c a a a g a g c t g g g c a g c c a g a t c c t g a a a g a a c a c c c c g t g g a a a a c a c c c a g c t g c a g a a g t t t c t c g a c c c g t c g g t c t a g g a c t t t c t t g t g g 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

3120 810 815 820 825 EKLYLYYLQNGRDMYVDQEL Cas9 810 815 820 825 EKLYLYYLQNGRDMYVDQEL Cas9 EKLYLYYLQNGRDMYVDQEL LLQVQVVQLIPPIHVHVLFQ c g a g a a g c t g t a c c t g t a c t a c c t g c a g a a t g g g c g g g a t a t g t a c g t g g a c c a g g a a c t g c t c t t c g a c a t g g a c a t g a t g g a c g t c t t a c c c g 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

3180 830 835 840 845 DINRLSDYDVDHIVPQSFLK Cas9 830 835 840 845 DINRLSDYDVDHIVPQSFLK Cas9 DINRLSDYDVDHIVPQSFLK VDVPQGVVIHVMDHRLAKQL g g a c a t c a a c c g g c t g t c c g a c t a c g a t g t g g a c c 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 c c t g t a g t t g g c c g a c a g g c t g a t g c t a c a c c t g g 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

3240 850 855 860 865 DDSIDNKVLTRSDKNRGKSD Cas9 850 855 860 865 DDSIDNKVLTRSDKNRGKSD Cas9 DDSIDNKVLTRSDKNRGKSD VVGDVVLHQGSAVLVPALAV g g a c g a c t c c a t c g a c a a c a a g g t g c t g a c c a g a a 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 c t g c t g a g g t a g c t g t t g t t c c a c g a c t g g t c t t 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

3300 870 875 880 885 NVPSEEVVKKMKNYWRQLLN Cas9 870 875 880 885 NVPSEEVVKKMKNYWRQLLN Cas9 NVPSEEVVKKMKNYWRQLLN VHGGFLDHLLHLVVPPLQQV c a a c g t g c c c t c c g a a g a g g t c g t g a a g a a g a t g a 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 g t t g c a c g g g a g g c t t c t c c a g c a c t t c t t c t a c t 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

3360 890 895 900 905 AKLITQRKFDNLTKAERGGL Cas9 890 895 900 905 AKLITQRKFDNLTKAERGGL Cas9 AKLITQRKFDNLTKAERGGL GLQNGLSLEVIQGLGLSAAQ c g c c a a g c t g a t t a c c c a g a g a a a g t t c g a c a a t c t 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 c g g t t c g a c t a a t g g g t c t c t t t c a a g c t g t t a g 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

3420 910 915 920 925 SELDKAGFIKRQLVETRQIT Cas9 910 915 920 925 SELDKAGFIKRQLVETRQIT Cas9 SELDKAGFIKRQLVETRQIT AFQILGAEDLSLQHFGPLDC g a g c g a a c t g g a t a a g g c c g g c t t c a t c a a g a g a c a g c t g g t g g a a a c c c g g c a g a t c a c c t c g c t t g a c c t a t t c c g g c c g a a g t a g t t c t c t g 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

3480 930 935 940 945 KHVAQILDSRMNTKYDENDK Cas9 930 935 940 945 KHVAQILDSRMNTKYDENDK Cas9 KHVAQILDSRMNTKYDENDK LVHCLDQVGPHVSLVVLIVL a a a g c a c g t g g c a c a g a t c c t g g a c t c c c g g a t g a 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 t t t c g t g c a c c g t g t c t a g g a c c t g a g g g c c t a c t 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

3540 950 955 960 965 LIREVKVITLKSKLVSDFRK Cas9 950 955 960 965 LIREVKVITLKSKLVSDFRK Cas9 LIREVKVITLKSKLVSDFRK QDPFHFHDGQLGLQHGIEPL 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 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 c g a c t a g g c c c t t c a c t t t c a c t a g t g g g a c t t c a 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

3600 970 975 980 985 DFQFYKVREINNYHHAHDAY Cas9 970 975 980 985 DFQFYKVREINNYHHAHDAY Cas9 DFQFYKVREINNYHHAHDAY IELKVFHALDVVVVVGVVGV 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 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 a a a g g t c a a a a t g t t t c a c g c g c t c t a g t t g t 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

3660 990 995 1000 1005 LNAVVGTALIKKYPKLESEF Cas9 990 995 1000 1005 LNAVVGTALIKKYPKLESEF Cas9 LNAVVGTALIKKYPKLESEF QVGDHSGGQDFLVRLQFALE c c t g a a c g c c g t c g t g g g a a c c g c c c t g a t c a a a a 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 g g a c t t g c g g c a g c a c c c t t g g c g g g a c t a g t t t t 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

3720 1010 1015 1020 1025 VYGDYKVYDVRKMIAKSEQE Cas9 1010 1015 1020 1025 VYGDYKVYDVRKMIAKSEQE Cas9 VYGDYKVYDVRKMIAKSEQE HVAVVLHVVHPLHDGLALLF c g t g t a c g g c g a c t a c a a g g t g t a c g a c g t g c g g a 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 g c a c a t g c c g c t g a t g t t c c a c a t g c t g c a c g c c t 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

3780 1030 1035 1040 1045 IGKATAKYFFYSNIMNFFKT Cas9 1030 1035 1040 1045 IGKATAKYFFYSNIMNFFKT Cas9 IGKATAKYFFYSNIMNFFKT DALSGGLVEEVAVDHVKELG a a t c g g c a a g g c t a c c g c c a a g t a c t t c t t c t a c a 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 t t a g c c g t t c c g a t g g c g g t t c a t g a a g a a g a t g t 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

3840 1050 1055 1060 1065 EITLANGEIRKRPLIETNGE Cas9 1050 1055 1060 1065 EITLANGEIRKRPLIETNGE Cas9 EITLANGEIRKRPLIETNGE LNGQGVALDPLPRQDLCVAF c g a g a t t a c c c t g g c c a a c g g c g a g a t c c g g a a g c 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 g c t c t a a t g g g a c c g g t t g c c g c t c t a g g c c t t c g 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

3900 1070 1075 1080 1085 TGEIVWDKGRDFATVRKVLS Cas9 1070 1075 1080 1085 TGEIVWDKGRDFATVRKVLS Cas9 TGEIVWDKGRDFATVRKVLS GPLDHPILAPIKGGHPFHQA a a c c g g g g a g a t c g t g t g g g a t a a g g g c c g g g a t t 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 t t g g c c c c t c t a g c a c a c c c t a t t c c c g g c c c t a a 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

3960 1090 1095 1100 1105 MPQVNIVKKTEVQTGGFSKE Cas9 1090 1095 1100 1105 MPQVNIVKKTEVQTGGFSKE Cas9 MPQVNIVKKTEVQTGGFSKE HGLHIDHFLGLHLCAAEAFL c a t g c c c c a a g t g a a t a t c g t g a a a a a g a c c g a g g 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 a c g g g g t t c a c t t a t a g c a c t t t t t c t g g c t c c 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

4020 1110 1115 1120 1125 SILPKRNSDKLIARKKDWDP Cas9 1110 1115 1120 1125 SILPKRNSDKLIARKKDWDP Cas9 SILPKRNSDKLIARKKDWDP RDQGLPVAILQDGSLLVPVR g t c t a t c c t g c c c a a g a g g a a c a g c g a t a a g c t g a 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 c a g a t a g g a c g g g t t c t c c t t g t c g c t a t t c g a c t 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

4080 1130 1135 1140 1145 KKYGGFDSPTVAYSVLVVAK Cas9 1130 1135 1140 1145 KKYGGFDSPTVAYSVLVVAK Cas9 KKYGGFDSPTVAYSVLVVAK LLVAAEVAGGHGIRHQHHGF t a a g a a g t a c g g c g g c t t c g a c a g c c c c a c c g t g g 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 t t c t t c a t g c c g c c g a a g c t g t c g g g g t g g c a c c 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

4140 1150 1155 1160 1165 VEKGKSKKLKSVKELLGITI Cas9 1150 1155 1160 1165 VEKGKSKKLKSVKELLGITI Cas9 VEKGKSKKLKSVKELLGITI HFLALGLFQLTHFLQQPDGD a g t g g a a a a g g g c a a g t c c a a g a a a c t g a a g a g t g 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 t c a c c t t t t c c c g t t c a g g t t c t t t g a c t t c t c a c 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

4200 1170 1175 1180 1185 MERSSFEKNPIDFLEAKGYK Cas9 1170 1175 1180 1185 MERSSFEKNPIDFLEAKGYK Cas9 MERSSFEKNPIDFLEAKGYK HFSAAELLIGDVKQFGLAVF c a t g g a a a g a a g c a g c t t c g a g a a g a a t c c c a t c g a c t t t c t g g a a g c c a a g g g c t a c a a g t a c c t t t c t t c g t c g a a g c t c t t c t t a g g g t a g c 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

4260 1190 1195 1200 1205 EVKKDLIIKLPKYSLFELEN Cas9 1190 1195 1200 1205 EVKKDLIIKLPKYSLFELEN Cas9 EVKKDLIIKLPKYSLFELEN FHFLVQDDLQRLVGQELQFV a g a a g t g a a a a a g g a c c t g a t c a t c a a g c t g c c t a 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 t c t t c a c t t t t t c c t g g a c t a g t a g t t c g a c g g a t 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

4320 1210 1215 1220 1225 GRKRMLASAGELQKGNELAL Cas9 1210 1215 1220 1225 GRKRMLASAGELQKGNELAL Cas9 GRKRMLASAGELQKGNELAL APLSHQGRGAFQLLSVFQGQ c g g c c g g a a g a g a a t g c t g g c c t c t g c c g g c g a a c t 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 g g c c t t c t c t t a c g a c c g g a g a c g g c c g c t t g 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

4380 1230 1235 1240 1245 PSKYVNFLYLASHYEKLKGS Cas9 1230 1235 1240 1245 PSKYVNFLYLASHYEKLKGS Cas9 PSKYVNFLYLASHYEKLKGS GGFIHVEQVQGAVILLQLAG g c c c t c c a a a t a t g t g a a c t t c c t g t a c c t g g c c a 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 g g g a g g t t t a t a c a c t t g a a g g a c a t g g a c c g g t 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

4440 1250 1255 1260 1265 PEDNEQKQLFVEQHKHYLDE Cas9 1250 1255 1260 1265 PEDNEQKQLFVEQHKHYLDE Cas9 PEDNEQKQLFVEQHKHYLDE GLIILLFLQKHFLVLVVQVL c c c c g a g g a t a a t g a g c a g a a a c a g c t g t t t g t g g 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 g g g c t c c t a t t a c t c g t c t t t g t c g a c a a a c a c c 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

4500 1270 1275 1280 1285 IIEQISEFSKRVILADANLD Cas9 1270 1275 1280 1285 IIEQISEFSKRVILADANLD Cas9 IIEQISEFSKRVILADANLD DDLLDALEGLSHDQGVSIQV g a t c a t c g a g c a g a t c a g c g a g t t c t c c a a g a g a g t 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 t a g t a g c t c g t c t a g t c g c t c a a g a g g t t c t c t c 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

4560 1290 1295 1300 1305 KVLSAYNKHRDKPIREQAEN Cas9 1290 1295 1300 1305 KVLSAYNKHRDKPIREQAEN Cas9 KVLSAYNKHRDKPIREQAEN FHQGGVVLVPILGDSLLGLI c a a a g t g c t g t c c g c c t a c a a c a a g c a c c g g g a t a a g c c c a t c a g a g a g c a g g c c g a g a a g t t t c a c g a c a g g c g g a t g t t g t t c g t g g c c c t a t 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

4620 1310 1315 1320 1325 IIHLFTLTNLGAPAAFKYFD Cas9 1310 1315 1320 1325 IIHLFTLTNLGAPAAFKYFD Cas9 IIHLFTLTNLGAPAAFKYFD DDVQKGQGIQSGRGGELVKV t a t c a t c c a c c t g t t t a c c c t g a c c a a t c t g g g a g 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 a t a g t a g g t g g a c a a a t g g g a c t g g t t a g a c c c t c 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

4680 1330 1335 1340 1345 TTIDRKRYTSTKEVLDATLI Cas9 1330 1335 1340 1345 TTIDRKRYTSTKEVLDATLI Cas9 TTIDRKRYTSTKEVLDATLI GGDVPLPVGAGFLHQVGGQD c a c c a c c a t c g a c c g g a a g a g g t a c a c c a g c a c c a 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 g t g g t g g t a g c t g g c c t t c t c c a t g t g g t c g t g g t 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

4740 1350 1355 1360 1365 HQSITGLYETRIDLSQLGGD Cas9 1350 1355 1360 1365 HQSITGLYETRIDLSQLGGD Cas9 HQSITGLYETRIDLSQLGGD VLADGAQVLCPDVQRLQSAV c c a c c a g a g c a t c a c c g g c c t g t a c g a g a c a c g g a t c g a c c t g t c t c a g c t g g g a g g c g a g g t g g t c t c g t a g t g g c c g g a c a t g c t c t g t g c c t 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

4800 1 5 10 15 KRPAATKKAGQAKKKK DYKD Cas9 nucleoplasmin NLS (in frame with Cas9) KRPAATKKAGQAKKKK 1 DYKD Cas9 FLAG KRPAATKKAGQAKKKKDYKD LSRGGCLLSSLSLLLFIVFV c a a g c g a c c t g c c g c c a c a a a g a a g g c t g g a c a g g c t a a g a a g a a g a a a g a t t a c a a a g a g t t c g c t g g a c g g c g g t g t t t c t t c c g a c c t g t c c g a t t c t t c t t c t t t c t a a t g t t t c t

BamHI 4860 (in frame with nucleoplasmin NLS) DDDKGSGATNFSLLKQAGDV 5 DDDK GSG 1 5 10 ATNFSLLKQAGDV FLAG P2A DDDKGSGATNFSLLKQAGDV IVILSGACCVERQQFLGSID c g a t g a c g a t a a g g g a t c c g g c g c a a c a a a c t t c t c t c t g c t g a a a c a a g c c g g a g a t g t g c t a c t g c t a t t c c c t a g g c c g c g t t g t t t g a a g a g a g a c g a c t t t g t t c g g c c t c t a c a

4920 (in frame with nucleoplasmin NLS) EENPGPAKPLSQEESTLIER 15 EENPGP 5 10 15 AKPLSQEESTLIER P2A BSD EENPGPAKPLSQEESTLIER FLIRSRGLRQRLFFGGENFS c g a a g a g a a t c c t g g a c c g g c c a a g c c t t t g t c t c a a g a a g a a t c c a c c c t c a t t g a a a g g c t t c t c t t a g g a c c t g g c c g g t t c g g a a a c a g a g t t c t t c t t a g g t g g g a g t a a c t t t c

4980 (in frame with nucleoplasmin NLS) ATATINSIPISEDYSVASAA 20 25 30 35 ATATINSIPISEDYSVASAA BSD ATATINSIPISEDYSVASAA CRSCDVADGDRFVVADGACS a g c a a c g g c t a c a a t c a a c a g c a t c c c c a t c t c t g a a g a c t a c a g c g t c g c c a g c g c a g c t c g t t g c c g a t g t t a g t t g t c g t a g g g g t a g a g a c t t c t g a t g t c g c a g c g g t c g c g t c g

5040 (in frame with nucleoplasmin NLS) LSSDGRIFTGVNVYHFTGGP 40 45 50 55 LSSDGRIFTGVNVYHFTGGP BSD LSSDGRIFTGVNVYHFTGGP ERAVAADESTDIYIMKSPSR t c t c t c t a g c g a c g g c c g c a t c t t c a c t g g t g t c a a t g t a t a t c a t t t t a c t g g g g g a c c a g a g a g a t c g c t g c c g g c g t a g a a g t g a c c a c a g t t a c a t a t a g t a a a a t g a c c c c c t g g

5100 (in frame with nucleoplasmin NLS) CAELVVLGTAAAAAAGNLTC 60 65 70 75 CAELVVLGTAAAAAAGNLTC BSD CAELVVLGTAAAAAAGNLTC TCFEHHQASSSSRCSAVQST t t g t g c a g a a c t c g t g g t g c t g g g c a c t g c t g c t g c t g c g g c a g c t g g c a a c c t g a c t t g a a c a c g t c t t g a g c a c c a c g a c c c g t g a c g a c g a c g a c g c c g t c g a c c g t t g g a c t g a a c

5160 (in frame with nucleoplasmin NLS) IVAIGNENRGILSPCGRCRQ 80 85 90 95 IVAIGNENRGILSPCGRCRQ BSD IVAIGNENRGILSPCGRCRQ DDRDSILVPADQAGASPASL t a t c g t c g c g a t c g g a a a t g a g a a c a g g g g c a t c t t g a g c c c c t g c g g a c g g t g c c g a c a a t a g c a g c g c t a g c c t t t a c t c t t g t c c c c g t a g a a c t c g g g g a c g c c t g c c a c g g c t g t

5220 (in frame with nucleoplasmin NLS) VLLDLHPGIKAIVKDSDGQP 100 105 110 115 VLLDLHPGIKAIVKDSDGQP BSD VLLDLHPGIKAIVKDSDGQP HKEIQM g g t g c t t c t c g a t c t g c a t c c t g g g a t c a a a g c c a t a g t g a a g g a c a g t g a t g g a c a g c c c c a c g a a g a g c t a g a c g t a g g a c c c t a g t t t c g g t a t c a c t t c c t g t c a c t a c c t g t c g g

MluI 5280 (in frame with nucleoplasmin NLS) TAVGIRELLPSGYVWEG* 120 125 130 TAVGIRELLPSGYVWEG* BSD TAVGIRELLPSGYVWEG* g a c g g c a g t t g g g a t t c g t g a a t t g c t g c c c t c t g g t t a t g t g t g g g a g g g c t a a a c g c g c t g c c g t c a a c c c t a a g c a c t t a a c g a c g g g a g a c c a a t a c a c a c c c t c c c g a t t t g c g c

5340 WPRE t t a a g t c g a c a a t c a a c c t c t g g a t t a c a a a a t t t g t g a a a g a t t g a c t g g t a t t c t t a a a a t t c a g c t g t t a g t t g g a g a c c t a a t g t t t t a a a c a c t t t c t a a c t g a c c a t a a g a a t t

5400 WPRE A C A A C G A G G A A A A T G C G A T A C WPRE-R MPLYHAI ML c t a t g t t g c t c c t t t t a c g c t a t g t g g a t a c g c t g c t t t a a t g c c t t t g t a t c a t g c t a t g a t a c a a c g a g g a a a a t g c g a t a c a c c t a t g c g a c g a a a t t a c g g a a a c a t a g t a c g a t a

5460 WPRE ASRMAFIFSSLYKSWLLSLY LLPVWLSFSPPCINPGCCLF t g c t t c c c g t a t g g c t t t c a t t t t c t c c t c c t t g t a t a a a t c c t g g t t g c t g t c t c t t t a a c g a a g g g c a t a c c g a a a g t a a a a g a g g a g g a a c a t a t t t a g g a c c a a c g a c a g a g a a a t

5520 WPRE EELWPVVRQRGVVCTVFADA MRSCGPLSGNVAWCALCLLT t g a g g a g t t g t g g c c c g t t g t c a g g c a a c g t g g c g t g g t g t g c a c t g t g t t t g c t g a c g c a c t c c t c a a c a c c g g g c a a c a g t c c g t t g c a c c g c a c c a c a c g t g a c a c a a a c g a c t g c g

5580 WPRE TPTGWGIATTCQLLSGTFAF QPPLVGALPPPVSSFPGLSL a a c c c c c a c t g g t t g g g g c a t t g c c a c c a c c t g t c a g c t c c t t t c c g g g a c t t t c g c t t t t t g g g g g t g a c c a a c c c c g t a a c g g t g g t g g a c a g t c g a g g a a a g g c c c t g a a a g c g a a a

5640 WPRE PLPIATAELIAACLARCWTG SPSLLPRRNSSPPALPAAGQ c c c c c t c c c t a t t g c c a c g g c g g a a c t c a t c g c c g c c t g c c t t g c c c g c t g c t g g a c a g g g g g g g a g g g a t a a c g g t g c c g c c t t g a g t a g c g g c g g a c g g a a c g g g c g a c g a c c t g t c c

5700 WPRE *RGKR ARLLGTDNSVVLSGKSSSFP GLGCWALTIPWCCRGNHRPF g g c t c g g c t g t t g g g c a c t g a c a a t t c c g t g g t g t t g t c g g g g a a a t c a t c g t c c t t t c c c c g a g c c g a c a a c c c g t g a c t g t t a a g g c a c c a c a a c a g c c c c t t t a g t a g c a g g a a a g g

5760 WPRE (in frame with Factor Xa site) PQEGTNGGPNQAPRGEAVDR WLLACVATWILRGTSFCYVP LGCSPVLPPGFCAGRPSATS t t g g c t g c t c g c c t g t g t t g c c a c c t g g a t t c t g c g c g g g a c g t c c t t c t g c t a c g t c c c a a c c g a c g a g c g g a c a c a a c g g t g g a c c t a a g a c g c g c c c t g c a g g a a g a c g a t g c a g g g

SacII 5820 WPRE 1 RGEI Factor Xa site SALNPADLPSRGLLPALRPL LRPSIQRTFLPAACCRLCGL t t c g g c c c t c a a t c c a g c g g a c c t t c c t t c c c g c g g c c t g c t g c c g g c t c t g c g g c c t c t a a g c c g g g a g t t a g g t c g c c t g g a a g g a a g g g c g c c g g a c g a c g g c c g a g a c g c c g g a g a

5880 WPRE PRLRLRPQTSRISLWAASPR FRVFAFALRRVGSPFGPPPR t c c g c g t c t t c g c c t t c g c c c t c a g a c g a g t c g g a t c t c c c t t t g g g c c g c c t c c c c g c g a g g c g c a g a a g c g g a a g c g g g a g t c t g c t c a g c c t a g a g g g a a a c c c g g c g g a g g g g c g c

5940 RL* VDFKTNDLQGSCRS* t c g a c t t t a a g a c c a a t g a c t t a c a a g g c a g c t g t a g a t c t t a g c c a c t t t t t a a a a g a a a g c t g a a a t t c t g g t t a c t g a a t g t t c c g t c g a c a t c t a g a a t c g g t g a a a a a t t t t c t t

6000 3' LTR ( Δ U3) a a g g g g g g a c t g g a a g g g c t a a t t c a c t c c c a a c g a a g a c a a g a t c t g c t t t t t g c t t g t t t c c c c c c t g a c c t t c c c g a t t a a g t g a g g g t t g c t t c t g t t c t a g a c g a a a a a c g a a c a

6060 3' LTR ( Δ U3) a c t g g g t c t c t c t g g t t a g a c c a g a t c t g a g c c t g g g a g c t c t c t g g c t a a c t a g g g a a c t g a c c c a g a g a g a c c a a t c t g g t c t a g a c t c g g a c c c t c g a g a g a c c g a t t g a t c c c t t g

6120 3' LTR ( Δ U3) c c a c t g c t t a a g c c t c a a t a a a g c t t g c c t t g a g t g c t t c a a g t a g t g t g t g c c c g t c t g g g t g a c g a a t t c g g a g t t a t t t c g a a c g g a a c t c a c g a a g t t c a t c a c a c a c g g g c a g a c

6180 3' LTR ( Δ U3) t t g t g t g a c t c t g g t a a c t a g a g a t c c c t c a g a c c c t t t t a g t c a g t g t g g a a a a t c t c t a a c a c a c t g a g a c c a t t g a t c t c t a g g g a g t c t g g g a a a a t c a g t c a c a c c t t t t a g a g a

PmeI 6240 3' LTR ( Δ U3) bGH poly(A) signal G G A G C T G A C A C G G A A G A T BGH-rev a g c a g g g c c c g t t t a a a c c c g c t g a t c a g c c t c g a c t g t g c c t t c t a g t t g c c a g c c a t c t c g t c c c g g g c a a a t t t g g g c g a c t a g t c g g a g c t g a c a c g g a a g a t c a a c g g t c g g t a g

6300 bGH poly(A) signal 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 c c a c t c c c a c t g t c c t 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 g g t g a g g g t g a c a g g a

6360 bGH poly(A) signal 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 t c a t t c t a t t c t g g g 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 c a g t a a g a t a a g a c c c

6420 bGH poly(A) signal 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 c a a t a g c a g g c a t g c t g g 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 g t t a t c g t c c g t a c g a c c

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

6540 f1 ori t c c c c 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 a g g g g 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

6600 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

6660 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

6720 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 a 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 t c c c a c t a c c a a g t g c a t c

6780 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

6840 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 t 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 a g a t a a g a a a a c t

6900 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

6960 f1 ori SV40 promoter C C T T A C A C A C A G T C A A T C C C A pBABE 3' a t t t a a c g c g a a t t a a t t c t g t g g a a t g t g t g t c a g t t a g g g t g t g g a a a g t c c c c a g g c t a a a t t g c g c t t a a t t a a g a c a c c t t a c a c a c a g t c a a t c c c a c a c c t t t c a g g g g t c c g

7020 SV40 promoter t c c c c a g c a g g c a g a a g t a t g c a a a g c a t g c a t c t c a a t t a g t c a g c a a c c a g g t g t g g a a g g g g t c g t c c g t c t t c a t a c g t t t c g t a c g t a g a g t t a a t c a g t c g t t g g t c c a c a c c t

7080 SV40 promoter a a g t c c c c a g g c t c c c c a g c a g g c a g a a g t a t g c a a a g c a t g c a t c t c a a t t a g t c a g c a t t c a g g g g t c c g a g g g g t c g t c c g t c t t c a t a c g t t t c g t a c g t a g a g t t a a t c a g t c g t

7140 SV40 promoter SV40 ori a c c a t a g t c c c g c c c c t a a c t c c g c c c a t c c c g c c c c t a a c t c c g c c c a g t t c c g c c c a t t g g t a t c a g g g c g g g g a t t g a g g c g g g t a g g g c g g g g a t t g a g g c g g g t c a a g g c g g g t a

7200 SV40 promoter SV40 ori T A T T T A T G C A G A G G C C G A G G SV40pro-F t c t c c g c c c c a t g g c t g a c t a a t t t t t t t t a t t t a t g c a g a g g c c g a g g c c g c c t c t g c c a g a g g c g g g g t a c c g a c t g a t t a a a a a a a a t a a a t a c g t c t c c g g c t c c g g c g g a g a c g g

StuIAvrII 7260 SV40 promoter SV40 ori t c t g a g c t a t t c c a g a a g t a g t g a g g a g g c t t t t t t g g a g g c c t a g g c t t t t g c a a a a a g a g a c t c g a t a a g g t c t t c a t c a c t c c t c c g a a a a a a c c t c c g g a t c c g a a a a c g t t t t t c

7320 EM7 promoter c t c c c g g g a g c t t g t a t a t c c a t t t t c g g a t c t g a t c a g c a c g t g t t g a c a a t t a a t c a t g a g g g c c c t c g a a c a t a t a g g t a a a a g c c t a g a c t a g t c g t g c a c a a c t g t t a a t t a g t a

7380 1 MAK EM7 promoter BleoR c g g c a t a g t a t a t c g g c a t a g t a t a a t a c g a c a a g g t g a g g a a c t a a a c c a t g g c c a a g t g c c g t a t c a t a t a g c c g t a t c a t a t t a t g c t g t t c c a c t c c t t g a t t t g g t a c c g g t t c a

7440 5 10 15 20 LTSAVPVLTARDVAGAVEFW BleoR t g a c c a g t g c c g t t c c g g t g c t c a c c g c g c g c g a c g t c g c c g g a g c g g t c g a g t t c t g g a a c t g g t c a c g g c a a g g c c a c g a g t g g c g c g c g c t g c a g c g g c c t c g c c a g c t c a a g a c c t

SgrAI 7500 25 30 35 40 TDRLGFSRDFVEDDFAGVVR BleoR c c g a c c g g c t c g g g t t c t c c c g g g a c t t c g t g g a g g a c g a c t t c g c c g g t g t g g t c c g g g g g c t g g c c g a g c c c a a g a g g g c c c t g a a g c a c c t c c t g c t g a a g c g g c c a c a c c a g g c c c

7560 45 50 55 60 DDVTLFISAVQDQVVPDNTL BleoR a c g a c g t g a c c c t g t t c a t c a g c g c g g t c c a g g a c c a g g t g g t g c c g g a c a a c a c c c t g g t g c t g c a c t g g g a c a a g t a g t c g c g c c a g g t c c t g g t c c a c c a c g g c c t g t t g t g g g a c c

7620 65 70 75 80 AWVWVRGLDELYAEWSEVVS BleoR c c t g g g t g t g g g t g c g c g g c c t g g a c g a g c t g t a c g c c g a g t g g t c g g a g g t c g t g t c c a g g a c c c a c a c c c a c g c g c c g g a c c t g c t c g a c a t g c g g c t c a c c a g c c t c c a g c a c a g g t

FseI 7680 85 90 95 100 TNFRDASGPAMTEIGEQPWG BleoR c g a a c t t c c g g g a c g c c t c c g g g c c g g c c a t g a c c g a g a t c g g c g a g c a g c c g t g g g g g c g c t t g a a g g c c c t g c g g a g g c c c g g c c g g t a c t g g c t c t a g c c g c t c g t c g g c a c c c c c g

7740 105 110 115 120 REFALRDPAGNCVHFVAEEQ BleoR g g g a g t t c g c c c t g c g c g a c c c g g c c g g c a a c t g c g t g c a c t t c g t g g c c g a g g a g c a g g c c c t c a a g c g g g a c g c g c t g g g c c g g c c g t t g a c g c a c g t g a a g c a c c g g c t c c t c g t c c

7800 125 D* BleoR a c t g a c a c g t g c t a c g a g a t t t c g a t t c c a c c g c c g c c t t c t a t g a a a g g t t g g g c t t c g t g a c t g t g c a c g a t g c t c t a a a g c t a a g g t g g c g g c g g a a g a t a c t t t c c a a c c c g a a g c

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

7920 SV40 poly(A) signal C G T T A T C G T SV40pA-R t c t t c g c c c a c c c c a a c t t g t t t a t t g c a g c t t a t a a t g g t t a c a a a t a a a g c a a t a g c a a g a a g c g g g t g g g g t t g a a c a a a t a a c g t c g a a t a t t a c c a a t g t t t a t t t c g t t a t c g t

7980 SV40 poly(A) signal G T G G T T T G T C C A A A C EBV-rev A G T G T T T A A A G SV40pA-R t c a c a a a t t t c a c a a a t a a a g c a t t t t t t t c a c t g c a t t c t a g t t g t g g t t t g t c c a a a c a g t g t t t a a a g t g t t t a t t t c g t a a a a a a a g t g a c g t a a g a t c a a c a c c a a a c a g g t t t g

BstZ17I 8040 SV40 poly(A) signal T C A T C EBV-rev t c a t c a a t g t a t c t t a t c a t g t c t g t a t a c c g t c g a c c t c t a g c t a g a g c t t g g c g t a a t a g t a g t t a c a t a g a a t a g t a c a g a c a t a t g g c a g c t g g a g a t c g a t c t c g a a c c g c a t t a

8100 -10 M13 rev lac operator lac promoter C A G T A T C G A C A A A G G A C M13 Reverse G G A C A C A C T T T A A C A A T A G G C G A M13/pUC Reverse c a t g g t c a t a g c t g t t t c c t g t g t g a a a t t g t t a t c c g c t c a c a a t t c c a c a c a a c a t a c g t a c c a g t a t c g a c a a a g g a c a c a c t t t a a c a a t a g g c g a g t g t t a a g g t g t g t t g t a t g

8160 -35 lac promoter CAP binding site g a g c c g g a a g c a t a a a g t g t a a a g c c t g g g g t g c c t a a t g a g t g a g c t a a c t c a c a t t a a c t c g g c c t t c g t a t t t c a c a t t t c g g a c c c c a c g g a t t a c t c a c t c g a t t g a g t g t a a t t

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

8280 G A G C G L4440 g a a t c g g c c a a c g c g c g g g g a g a g g c g g t t t g c g t a t t g g g c g c t c t t c c g c t t c c t c g c c t t a g c c g g t t g c g c g c c c c t c t c c g c c a a a c g c a t a a c c c g c g a g a a g g c g a a g g a g c g

8340 A G T G A C T G A G C G A L4440 t c a c t g a c t c g c t g c g c t c g g t c g t t c g g c t g c g g c g a g c g g t a t c a g c t c a c t c a a a g g a g t g a c t g a g c g a c g c g a g c c a g c a a g c c g a c g c c g c t c g c c a t a g t c g a g t g a g t t t c c

PciI 8400 c g g t a a t a c g g t t a t c c a c a g a a t c a g g g g a t a a c g c a g g a a a g a a c a t g t g a g c a a a a g g c c a t t a t g c c a a t a g g t g t c t t a g t c c c c t a t t g c g t c c t t t c t t g t a c a c t c g t t t t c

8460 ori g c c a g c a a a a g g c c a g g a a c c g t a a a a a g g c c g c g t t g c t g g c g t t t t t c c a t a g g c t c c c g g t c g t t t t c c g g t c c t t g g c a t t t t t c c g g c g c a a c g a c c g c a a a a a g g t a t c c g a g g

8520 ori g c c c c c c t g a c g a g c a t c a c a a a a a t c g a c g c t c a a g t c a g a g g t g g c g a a a c c c g a c a g c g g g g g g a c t g c t c g t a g t g t t t t t a g c t g c g a g t t c a g t c t c c a c c g c t t t g g g c t g t c

8580 ori T T T C T A T G G T C C G C A A A G G G pBR322ori-F g a c t a t a a a g a t a c c a g g c g t t t c c c c c t g g a a g c t c c c t c g t g c g c t c t c c t g t t c c g a c t g a t a t t t c t a t g g t c c g c a a a g g g g g a c c t t c g a g g g a g c a c g c g a g a g g a c a a g g c t

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

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

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

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

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

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

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

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

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

9180 a a a g g a t c t t c a c c t a g a t c c t t t t a a a t t a a a a a t g a a g t t t t a a a t c a a t c t a a a g t a t t t c c t a g a a g t g g a t c t a g g a a a a t t t a a t t t t t a c t t c a a a a t t t a g t t a g a t t t c a t

9240 285 280 *WHKILSAGIEA AmpR *WHKILSAGIEA t a t a t g a g t a a a c t t g g t c t g a c a g t t a c c a a t g c t t a a t c a g t g a g g c a c c t a t c t c a g a t a t a c t c a t t t g a a c c a g a c t g t c a a t g g t t a c g a a t t a g t c a c t c c g t g g a t a g a g t c

9300 275 270 265 260 IQRNREDMTAQSGTTYIVVI AmpR IQRNREDMTAQSGTTYIVVI c g a t c t g t c t a t t t c g t t c a t c c a t a g t t g c c t g a c t c c c c g t c g t g t a g a t a a c t a c g a g c t a g a c a g a t a a a g c a a g t a g g t a t c a a c g g a c t g a g g g g c a g c a c a t c t a t t g a t g c t

9360 255 250 245 240 RSPKGDPGLAAIIGRSGREG AmpR RSPKGDPGLAAIIGRSGREG t a c g g g a g g g c t t a c c a t c t g g c c c c a g t g c t g c a a t g a t a c c g c g a g a c c c a c g c t c a c a t g c c c t c c c g a a t g g t a g a c c g g g g t c a c g a c g t t a c t a t g g c g c t c t g g g t g c g a g t g

9420 235 230 225 220 AGSKDAIFWGAPLASRLLPG AmpR AGSKDAIFWGAPLASRLLPG c g g c t c c a g a t t t a t c a g c a a t a a a c c a g c c a g c c g g a a g g g c c g a g c g c a g a a g t g g t c g c c g a g g t c t a a a t a g t c g t t a t t t g g t c g g t c g g c c t t c c c g g c t c g c g t c t t c a c c a g

9480 215 210 205 200 AVKDAEMWDILQQRSALTLL AmpR AVKDAEMWDILQQRSALTLL c t g c a a c t t t a t c c g c c t c c a t c c a g t c t a t t a a t t g t t g c c g g g a a g c t a g a g t a a g t a g a c g t t g a a a t a g g c g g a g g t a g g t c a g a t a a t t a a c a a c g g c c c t t c g a t c t c a t t c a t

FspI 9540 195 190 185 180 EGTLLKRLTTAMAVPMTTDR AmpR EGTLLKRLTTAMAVPMTTDR g t t c g c c a g t t a a t a g t t t g c g c a a c g t t g t t g c c a t t g c t a c a g g c a t c g t g g t g t c a c c a a g c g g t c a a t t a t c a a a c g c g t t g c a a c a a c g g t a a c g a t g t c c g t a g c a c c a c a g t g

9600 175 170 165 160 EDNPIAENLEPEWRDLRTVH AmpR EDNPIAENLEPEWRDLRTVH g c t c g t c g t t t g g t a t g g c t t c a t t c a g c t c c g g t t c c c a a c g a t c a a g g c g a g t t a c a t c g a g c a g c a a a c c a t a c c g a a g t a a g t c g a g g c c a a g g g t t g c t a g t t c c g c t c a a t g t a

9660 155 150 145 140 DGMNHLFATLEKPGGITTLL AmpR DGMNHLFATLEKPGGITTLL g a t c c c c c a t g t t g t g c a a a a a a g c g g t t a g c t c c t t c g g t c c t c c g a t c g t t g t c a g a a c t a g g g g g t a c a a c a c g t t t t t t c g c c a a t c g a g g a a g c c a g g a g g c t a g c a a c a g t c t t

9720 135 130 125 120 LNAATNDSMTIAASCLERVT AmpR LNAATNDSMTIAASCLERVT g t a a g t t g g c c g c a g t g t t a t c a c t c a t g g t t a t g g c a g c a c t g c a t a a t t c t c t t a c t g c a t t c a a c c g g c g t c a c a a t a g t g a g t a c c a a t a c c g t c g t g a c g t a t t a a g a g a a t g a c

9780 115 110 105 100 MGDTLHKETVPSYEVLDNQS AmpR MGDTLHKETVPSYEVLDNQS t c a t g c c a t c c g t a a g a t g c t t t t c t g t g a c t g g t g a g t a c t c a a c c a a g t c a t t c t g a g a g t a c g g t a g g c a t t c t a c g a a a a g a c a c t g a c c a c t c a t g a g t t g g t t c a g t a a g a c t c

9840 95 90 85 80 YHIRRGLQEQGADIRSLVAG AmpR A T A A T A C C G C G C Amp-R YHIRRGLQEQGADIRSLVAG a a t a g t g t a t g c g g c g a c c g a g t t g c t c t t g c c c g g c g t c a a t a c g g g a t a a t a c c g c g c t t a t c a c a t a c g c c g c t g g c t c a a c g a g a a c g g g c c g c a g t t a t g c c c t a t t a t g g c g c g

9900 75 70 65 60 CLLVKFTSMMPFREEPRFSE AmpR C A C A T A G C Amp-R CLLVKFTSMMPFREEPRFSE c a c a t a g c a g a a c t t t a a a a g t g c t c a t c a t t g g a a a a c g t t c t t c g g g g c g a a a a c t c t g t g t a t c g t c t t g a a a t t t t c a c g a g t a g t a a c c t t t t g c a a g a a g c c c c g c t t t t g a g a

9960 55 50 45 40 LIKGSNLDLEIYGVRAGLQD AmpR LIKGSNLDLEIYGVRAGLQD c a a g g a t c t t a c c g c t g t t g a g a t c c a g t t c g a t g t a a c c c a c t c g t g c a c c c a a c t g a t g t t c c t a g a a t g g c g a c a a c t c t a g g t c a a g c t a c a t t g g g t g a g c a c g t g g g t t g a c t a

10,020 35 30 25 EADKVKVLTEPH 20 signal sequence AFVPLCFA AmpR EADKVKVLTEPHAFVPLCFA c t t c a g c a t c t t t t a c t t t c a c c a g c g t t t c t g g g t g a g c a a a a a c a g g a a g g c a a a a t g g a a g t c g t a g a a a a t g a a a g t g g t c g c a a a g a c c c a c t c g t t t t t g t c c t t c c g t t t t a c

10,080 15 10 5 1 signal sequence AFFPILAVRFHQISM AmpR AmpR promoter AFFPILAVRFHQISM c c g c a a a a a a g g g a a t a a g g g c g a c a c g g a a a t g t t g a a t a c t c a t a c t c t t c c t t t t t c g g c g t t t t t t c c c t t a t t c c c g c t g t g c c t t t a c a a c t t a t g a g t a t g a g a a g g a a a a a g

SspI 10,140 AmpR promoter a a t a t t a t t g a a g c a t t t a t c a g g g t t a t t g t c t c a t g a g c g g a t a c a t a t t t g a a t g t a t t a t a a t a a c t t c g t a a a t a g t c c c a a t a a c a g a g t a c t c g c c t a t g t a t a a a c t t a c a t

10,200 AmpR promoter t t t a g a a a a a t a a a c a a a t a g g g g t t c c g c g c a c a t t t c c c c g a a a a g t g c c a c c t g a c g a a a t c t t t t t a t t t g t t t a t c c c c a a g g c g c g t g t a a a g g g g c t t t t c a c g g t g g a c t g c

SgrDI 10,260 A T G T T A G A C G A G A C T pRS-marker t c g a c g g a t c g g g a g a t c t c c c g a t c c c c t a t g g t g c a c t c t c a g t a c a a t c t g c t c t g a a g c t g c c t a g c c c t c t a g a g g g c t a g g g g a t a c c a c g t g a g a g t c a t g t t a g a c g a g a c t

10,320 A C G G C pRS-marker t g c c g c a t a g t t a a g c c a g t a t c t g c t c c c t g c t t g t g t g t t g g a g g t c g c t g a g t a g t g a c g g c g t a t c a a t t c g g t c a t a g a c g a g g g a c g a a c a c a c a a c c t c c a g c g a c t c a t c a c

10,380 c g c g a g c a a a a t t t a a g c t a c a a c a a g g c a a g g c t t g a c c g a c a a t t g c a t g a a g a a t c t g c g c t c g t t t t a a a t t c g a t g t t g t t c c g t t c c g a a c t g g c t g t t a a c g t a c t t c t t a g a

10,440 g c t t a g g g t t a g g c g t t t t g c g c t g c t t c g c g a t g t a c g g g c c a g a t a t a c g c g c g c g t t c g a a t c c c a a t c c g c a a a a c g c g a c g a a g c g c t a c a t g c c c g g t c t a t a t g c g c g c g c a a

SpeI 10,500 CMV enhancer g a c a t t g a t t a t t g a c t a g t t a t t a a t a g t a a t c a a t t a c g g g g t c a t t a g t t c a t a g c c c t g t a a c t a a t a a c t g a t c a a t a a t t a t c a t t a g t t a a t g c c c c a g t a a t c a a g t a t c g g

10,560 CMV enhancer c a t a t a t g g a g t t c c g c g t t a c a t a a c t t a c g g t a a a t g g c c c g c c t g g c t g a c c g c c c a g t a t a t a c c t c a a g g c g c a a t g t a t t g a a t g c c a t t t a c c g g g c g g a c c g a c t g g c g g g t

10,620 CMV enhancer 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 a t g a c g t a t g t t c c c a t a g t a a c g c c a a t a g g g a 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 t a c t g c a t a c a a g g g t a t c a t t g c g g t t a t c c c t

10,680 CMV enhancer 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 t c 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 a g

NdeI 10,740 CMV enhancer 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 c t 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 g a

SnaBI 10,800 CMV enhancer g g c a t t a t g c c c a g t a c a t g a c c t t a t g g g a c t t t c c t a c t t g g c a g t a c a t c t a c g t a t c c g t a a t a c g g g t c a t g t a c t g g a a t a c c c t g a a a g g a t g a a c c g t c a t g t a g a t g c a t a

10,860 CMV enhancer CMV promoter t a g t c a t c g c t a t t a c c a t g g t g a t g c g g t t t t g g c a g t a c a t c a a t g g g c g t g g a t a g c a t c a g t a g c g a t a a t g g t a c c a c t a c g c c a a a a c c g t c a t g t a g t t a c c c g c a c c t a t c g

10,920 CMV promoter g g t t t g a c t c a c g g g g a t t t c c a a g t c t c c a c c c c a t t g a c g t c a a t g g g a g t t t g t t t t c c a a a c t g a g t g c c c c t a a a g g t t c a g a g g t g g g g t a a c t g c a g t t a c c c t c a a a c a a a a

10,980 CMV promoter C G C A A A CMV-F g g c a c c a a a a t c a a c g g g a c t t t c c a a a a t g t c g t a a c a a c t c c g c c c c a t t g a c g c a a a c c g t g g t t t t a g t t g c c c t g a a a g g t t t t a c a g c a t t g t t g a g g c g g g g t a a c t g c g t t t

11,040 CMV promoter 5' LTR (truncated) T G G G C G G T A G G C G T G CMV-F t g g g c g g t a g g c g t g t a c g g t g g g a g g t c t a t a t a a g c a g c g c g t t t t g c c t g t a c t g g g a c c c g c c a t c c g c a c a t g c c a c c c t c c a g a t a t a t t c g t c g c g c a a a a c g g a c a t g a c c c

11,100 5' LTR (truncated) t c t c t c t g g t t a g a c c a g a t c t g a g c c t g g g a g c t c t c t g g c t a a c t a g g g a a c c c a c t g a g a g a g a c c a a t c t g g t c t a g a c t c g g a c c c t c g a g a g a c c g a t t g a t c c c t t g g g t g a c

11,160 5' LTR (truncated) c t t a a g c c t c a a t a a a g c t t g c c t t g a g t g c t t c a a g t a g t g t g t g c c c g t c t g t t g t g t g a a t t c g g a g t t a t t t c g a a c g g a a c t c a c g a a g t t c a t c a c a c a c g g g c a g a c a a c a c a

11,220 5' LTR (truncated) g a c t c t g g t a a c t a g a g a t c c c t c a g a c c c t t t t a g t c a g t g t g g a a a a t c t c t a g c a g t c t g a g a c c a t t g a t c t c t a g g g a g t c t g g g a a a a t c a g t c a c a c c t t t t a g a g a t c g t c a

KasINarISfoIPluTI 11,280 HIV-1 Ψ g g c g c c c g a a c a g g g a c t t g a a a g c g a a a g g g a a a c c a g a g g a g c t c t c t c g a c g c a g g a c c g c g g g c t t g t c c c t g a a c t t t c g c t t t c c c t t t g g t c t c c t c g a g a g a g c t g c g t c c t

11,340 HIV-1 Ψ c t c g g c t t g c t g a a g c g c g c a c g g c a a g a g g c g a g g g g c g g c g a c t g g t g a g t a c g c c a a g a g c c g a a c g a c t t c g c g c g t g c c g t t c t c c g c t c c c c g c c g c t g a c c a c t c a t g c g g t t

11,400 HIV-1 Ψ a a a t t t t g a c t a g c g g a g g c t a g a a g g a g a g a g a t g g g t g c g a g a g c g t c a g t a t t a a g c t t t a a a a c t g a t c g c c t c c g a t c t t c c t c t c t c t a c c c a c g c t c t c g c a g t c a t a a t t c g

11,460 g g g g g a g a a t t a g a t c g c g a t g g g a a a a a a t t c g g t t a a g g c c a g g g g g a a a g a a a a a a t c c c c c t c t t a a t c t a g c g c t a c c c t t t t t t a a g c c a a t t c c g g t c c c c c t t t c t t t t t t a

11,520 a t a a a t t a a a a c a t a t a g t a t g g g c a a g c a g g g a g c t a g a a c g a t t c g c a g t t a a t c c t g t a t t t a a t t t t g t a t a t c a t a c c c g t t c g t c c c t c g a t c t t g c t a a g c g t c a a t t a g g a c

11,580 g c c t g t t a g a a a c a t c a g a a g g c t g t a g a c a a a t a c t g g g a c a g c t a c a a c c a t c c c t t c c g g a c a a t c t t t g t a g t c t t c c g a c a t c t g t t t a t g a c c c t g t c g a t g t t g g t a g g g a a g

11,640 a g a c a g g a t c a g a a g a a c t t a g a t c a t t a t a t a a t a c a g t a g c a a c c c t c t a t t g t g t g c t c t g t c c t a g t c t t c t t g a a t c t a g t a a t a t a t t a t g t c a t c g t t g g g a g a t a a c a c a c g

11,700 a t c a a a g g a t a g a g a t a a a a g a c a c c a a g g a a g c t t t a g a c a a g a t a g a g g a a g a g c a a a t a g t t t c c t a t c t c t a t t t t c t g t g g t t c c t t c g a a a t c t g t t c t a t c t c c t t c t c g t t t

NotI 11,760 a c a a a a g t a a g a c c a c c g c a c a g c a a g c g g c c g c t g a t c t t c a g a c c t g g a g g a g g a g a t t g t t t t c a t t c t g g t g g c g t g t c g t t c g c c g g c g a c t a g a a g t c t g g a c c t c c t c c t c t a

11,820 MRDNWRSELYKYKVVKIEPL a t g a g g g a c a a t t g g a g a a g t g a a t t a t a t a a a t a t a a a g t a g t a a a a a t t g a a c c a t t a t a c t c c c t g t t a a c c t c t t c a c t t a a t a t a t t t a t a t t t c a t c a t t t t t a a c t t g g t a a t

11,880 GVAPTKAKRRVVQREKRAVG g g a g t a g c a c c c a c c a a g g c a a a g a g a a g a g t g g t g c a g a g a g a a a a a a g a g c a g t g g g a c c t c a t c g t g g g t g g t t c c g t t t c t c t t c t c a c c a c g t c t c t c t t t t t t c t c g t c a c c c t

11,940 RRE IGALFLGFLGAAGSTMGAAS a t a g g a g c t t t g t t c c t t g g g t t c t t g g g a g c a g c a g g a a g c a c t a t g g g c g c a g c g t c a t a t c c t c g a a a c a a g g a a c c c a a g a a c c c t c g t c g t c c t t c g t g a t a c c c g c g t c g c a g t

12,000 RRE MTLTVQARQLLSGIVQQQNN a t g a c g c t g a c g g t a c a g g c c a g a c a a t t a t t g t c t g g t a t a g t g c a g c a g c a g a a c a a t t a c t g c g a c t g c c a t g t c c g g t c t g t t a a t a a c a g a c c a t a t c a c g t c g t c g t c t t g t t a

BbvCI 12,060 RRE LLRAIEAQQHLLQLTVWGIK t t g c t g a g g g c t a t t g a g g c g c a a c a g c a t c t g t t g c a a c t c a c a g t c t g g g g c a t c a a g a a c g a c t c c c g a t a a c t c c g c g t t g t c g t a g a c a a c g t t g a g t g t c a g a c c c c g t a g t t c

12,120 RRE QLQARILAVERYLKDQQLLG c a g c t c c a g g c a a g a a t c c t g g c t g t g g a a a g a t a c c t a a a g g a t c a a c a g c t c c t g g g g g t c g a g g t c c g t t c t t a g g a c c g a c a c c t t t c t a t g g a t t t c c t a g t t g t c g a g g a c c c c

AleI 12,180 IWGCSGKLICTTAVPWNASW a t t t g g g g t t g c t c t g g a a a a c t c a t t t g c a c c a c t g c t g t g c c t t g g a a t g c t a g t t g g t a a a c c c c a a c g a g a c c t t t t g a g t a a a c g t g g t g a c g a c a c g g a a c c t t a c g a t c a a c c

12,240 SNKSLEQIWNHTTWMEWDRE a g t a a t a a a t c t c t g g a a c a g a t t t g g a a t c a c a c g a c c t g g a t g g a g t g g g a c a g a g a a t c a t t a t t t a g a g a c c t t g t c t a a a c c t t a g t g t g c t g g a c c t a c c t c a c c c t g t c t c t t

12,300 INNYTSLIHSLIEESQNQQE a t t a a c a a t t a c a c a a g c t t a a t a c a c t c c t t a a t t g a a g a a t c g c a a a a c c a g c a a g a a t a a t t g t t a a t g t g t t c g a a t t a t g t g a g g a a t t a a c t t c t t a g c g t t t t g g t c g t t c t t

12,360 1 5 10 15 KNEQELLELDKWASL WNWFN gp41 peptide KNEQELLELDKWASLWNWFN a a g a a t g a a c a a g a a t t a t t g g a a t t a g a t a a a t g g g c a a g t t t g t g g a a t t g g t t t a a c t t c t t a c t t g t t c t t a a t a a c c t t a a t c t a t t t a c c c g t t c a a a c a c c t t a a c c a a a t t g

12,420 (in frame with gp41 peptide) ITNWLWYIKLFIMIVGGLVG ITNWLWYIKLFIMIVGGLVG a t a a c a a a t t g g c t g t g g t a t a t a a a a t t a t t c a t a a t g a t a g t a g g a g g c t t g g t a g g t t a t t g t t t a a c c g a c a c c a t a t a t t t t a a t a a g t a t t a c t a t c a t c c t c c g a a c c a t c c a

12,480 (in frame with gp41 peptide) LRIVFAVLSIVNRVRQGYSP LRIVFAVLSIVNRVRQGYSP t t a a g a a t a g t t t t t g c t g t a c t t t c t a t a g t g a a t a g a g t t a g g c a g g g a t a t t c a c c a a a t t c t t a t c a a a a a c g a c a t g a a a g a t a t c a c t t a t c t c a a t c c g t c c c t a t a a g t g g t

12,540 (in frame with gp41 peptide) LSFQTHLPTPRGPDRPEGIE LSFQTHLPTPRGPDRPEGIE t t a t c g t t t c a g a c c c a c c t c c c a a c c c c g a g g g g a c c c g a c a g g c c c g a a g g a a t a g a a a a t a g c a a a g t c t g g g t g g a g g g t t g g g g c t c c c c t g g g c t g t c c g g g c t t c c t t a t c t t

12,600 (in frame with gp41 peptide) EEGGERDRDRSIRLVNGSAL EEGGERDRDRSIRLVNGSAL g a a g a a g g t g g a g a g a g a g a c a g a g a c a g a t c c a t t c g a t t a g t g a a c g g a t c g g c a c t g c t t c t t c c a c c t c t c t c t c t g t c t c t g t c t a g g t a a g c t a a t c a c t t g c c t a g c c g t g a c

12,660 (in frame with gp41 peptide) RAPILQTNGSIHPQF* cPPT/CTS RAPILQTNGSIHPQF* c 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 a g t a t t c a t c c a c a a t t t t a a a a g a a a a g g g g g g 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 t c a t a a g t a g g t g t t a a a a t t t t c t t t t c c c c c

12,720 cPPT/CTS g a t t g g g g g g t a c a g t g c a g g g g a a a g a a t a g t a g a c a t a a t a g c a a c a g a c a t a c a a a c c t a a c c c c c c a t g t c a c g t c c c c t t t c t t a t c a t c t g t a t t a t c g t t g t c t g t a t g t t t g

12,780 cPPT/CTS t a a a g a a t t a c a a a a a c a a a t t a c a a a a a t t c a a a a t t t t c g g g t t t a t t a c a g g g a c a g a t t t c t t a a t g t t t t t g t t t a a t g t t t t t a a g t t t t a a a a g c c c a a a t a a t g t c c c t g t c

PacIAcc65IKpnI 3ʹ 5ʹ 12,810 c a g a g a t c c a g t t t g g t t a a t t a a g g t a c c g t c t c t a g g t c a a a c c a a t t a a t t c c a t 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.