Addgene pCMV-T7-ABE8e-nSpRY-P2A-EGFP (KAC1069) Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pCMV-T7-ABE8e-nSpRY-P2A-EGFP (KAC1069) Sequencing Result


Map View

SpeI (8799) MluI (8778) NruI (8758) MfeI (8711) pRS-marker (8594 .. 8613) BspDI * - ClaI * (8561) SgrDI (8544) SspI (8427) Amp-R (8173 .. 8192) TsoI (8022) PvuI (7993) FspI (7845) BsaI (7684) pBR322ori-F (6871 .. 6890) DrdI (6838) PciI (6730) L4440 (6620 .. 6637) BspQI - SapI (6614) CAP binding site lac promoter M13/pUC Reverse (6402 .. 6424) lac operator M13 Reverse (6389 .. 6405) M13 rev BbsI (6283) BGH-rev (6088 .. 6105) PmeI (6072) 6xHis AgeI (6042) BsrGI (5999) EGFP-C (5943 .. 5964) EXFP-R (5596 .. 5615) P2A BsmBI - Esp3I (5258) EcoRI (5194) BsmI (4697) NdeI (1) SnaBI (107) NcoI (127) CMV-F (286 .. 306) NotI - SacII (370) T7 (377 .. 396) T7 promoter SV40 NLS MreI - SgrAI (733) BglII (1307) PspOMI (2412) ApaI (2416) PasI (2826) EcoRV (2910) PmlI (3846) KflI (4434) BsaBI * (4555) pCMV-T7-ABE8e-nSpRY-P2A-EGFP (KAC1069) 9033 bp

Sequence View

NdeI 5ʹ 3ʹ 60 CMV enhancer a t a t g c c a a g t a c g c c c c c t a t t g a c g t c a a t g a c g g t a a a t g g c c c g c c t g g c a t t a t g t a t a c g g t t c a t g c g g g g g a t a a c t g c a g t t a c t g c c a t t t a c c g g g c g g a c c g t a a t a c

SnaBI 120 CMV enhancer c c c a g t a c a t g a c c t t a t g g g a c t t t c c t a c t t g g c a g t a c a t c t a c g t a t t a g t c a t c g g g g t c a t g t a c t g g a a t a c c c t g a a a g g a t g a a c c g t c a t g t a g a t g c a t a a t c a g t a g c

NcoI 180 CMV enhancer CMV promoter 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 g g t t t g a c t 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 c c a a a c t g a

240 CMV promoter 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 g g c a c c a a 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 c c g t g g t t t

300 CMV promoter C G C A A A T G G G C G G T A CMV-F 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 t g g g c g g t a 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 a c c c g c c a t

360 CMV promoter G G C G T G CMV-F 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 a g c t g g t t t a g t g a a c c g t c a g a t c c g c 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 t c g a c c a a a t c a c t t g g c a g t c t a g g c g a

SacIINotI 420 T7 promoter T A A T A C G A C T C A C T A T A G G G T7 MKRT a g a g a t c c g c g g c c g c t a a t a c g a c t c a c t a t a g g g a g a g c c g c c a c c a t g a a a c g g a c a t c t c t a g g c g c c g g c g a t t a t g c t g a g t g a t a t c c c t c t c g g c g g t g g t a c t t t g c c t g t

480 130 PKKKRKV (in frame with SV40 NLS) SEVEF SV40 NLS ADGSEFESPKKKRKVSEVEF g c c g a c g g a a g c g a g t t c g a g t c a c c a a a g a a g a a g c g g a a a g t c t c t g a g g t g g a g t t t c g g c t g c c t t c g c t c a a g c t c a g t g g t t t c t t c t t c g c c t t t c a g a g a c t c c a c c t c a a a

540 (in frame with SV40 NLS) SHEYWMRHALTLAKRARDER SHEYWMRHALTLAKRARDER t c c c a c g a g t a c t g g a t g a g a c a t g c c c t g a c c c t g g c c a a g a g g g c a c g c g a t g a g a g g a g g g t g c t c a t g a c c t a c t c t g t a c g g g a c t g g g a c c g g t t c t c c c g t g c g c t a c t c t c c

600 (in frame with SV40 NLS) EVPVGAVLVLNNRVIGEGWN EVPVGAVLVLNNRVIGEGWN g a g g t g c c t g t g g g a g c c g t g c t g g t g c t g a a c a a t a g a g t g a t c g g c g a g g g c t g g a a c c t c c a c g g a c a c c c t c g g c a c g a c c a c g a c t t g t t a t c t c a c t a g c c g c t c c c g a c c t t g

660 (in frame with SV40 NLS) RAIGLHDPTAHAEIMALRQG RAIGLHDPTAHAEIMALRQG a g a g c c a t c g g c c t g c a c g a c c c a a c a g c c c a t g c c g a a a t t a t g g c c c t g a g a c a g g g c t c t c g g t a g c c g g a c g t g c t g g g t t g t c g g g t a c g g c t t t a a t a c c g g g a c t c t g t c c c g

720 (in frame with SV40 NLS) GLVMQNYRLIDATLYVTFEP GLVMQNYRLIDATLYVTFEP g g c c t g g t c a t g c a g a a c t a c a g a c t g a t t g a c g c c a c c c t g t a c g t g a c a t t c g a g c c t c c g g a c c a g t a c g t c t t g a t g t c t g a c t a a c t g c g g t g g g a c a t g c a c t g t a a g c t c g g a

SgrAIMreI 780 (in frame with SV40 NLS) CVMCAGAMIHSRIGRVVFGV CVMCAGAMIHSRIGRVVFGV t g c g t g a t g t g c g c c g g c g c c a t g a t c c a c t c t a g g a t c g g c c g c g t g g t g t t t g g c g t g a c g c a c t a c a c g c g g c c g c g g t a c t a g g t g a g a t c c t a g c c g g c g c a c c a c a a a c c g c a c

840 (in frame with SV40 NLS) RNSKRGAAGSLMNVLNYPGM RNSKRGAAGSLMNVLNYPGM a g g a a c a g c a a a a g a g g c g c c g c a g g c t c c c t g a t g a a c g t g c t g a a c t a c c c c g g c a t g t c c t t g t c g t t t t c t c c g c g g c g t c c g a g g g a c t a c t t g c a c g a c t t g a t g g g g c c g t a c

900 (in frame with SV40 NLS) NHRVEITEGILADECAALLC NHRVEITEGILADECAALLC a a t c a c c g c g t c g a a a t t a c c g a g g g a a t c c t g g c a g a t g a a t g t g c c g c c c t g c t g t g c t t a g t g g c g c a g c t t t a a t g g c t c c c t t a g g a c c g t c t a c t t a c a c g g c g g g a c g a c a c g

960 (in frame with SV40 NLS) DFYRMPRQVFNAQKKAQSSI DFYRMPRQVFNAQKKAQSSI g a t t t c t a c c g g a t g c c t a g a c a g g t g t t c a a t g c t c a g a a g a a g g c c c a g a g c t c c a t c c t a a a g a t g g c c t a c g g a t c t g t c c a c a a g t t a c g a g t c t t c t t c c g g g t c t c g a g g t a g

1020 (in frame with SV40 NLS) NSGGSSGGSSGSETPGTSES NSGGSSGGSSGSETPGTSES a a c t c c g g a g g a t c t a g c g g a g g c t c c t c t g g c t c t g a g a c a c c t g g c a c a a g c g a g a g c t t g a g g c c t c c t a g a t c g c c t c c g a g g a g a c c g a g a c t c t g t g g a c c g t g t t c g c t c t c g

1080 (in frame with SV40 NLS) ATPESSGGSSGGS 5 DKKYSIG Cas9 ATPESSGGSSGGSDKKYSIG *VLLVADA g c a a c a c c t g a a a g c a g c g g g g g c a g c a g c g g g g g g t c a g a c a a g a a g t a c a g c a t c g g c c g t t g t g g a c t t t c g t c g c c c c c g t c g t c g c c c c c c a g t c t g t t c t t c a t g t c g t a g c c g

1140 10 15 20 25 LAIGTNSVGWAVITDEYKVP Cas9 LAIGTNSVGWAVITDEYKVP QGDAGVRHAPGHDGVLVLHG c t g g c c a t c g g c a c c a a c t c t g t g g g c t g g g c c g 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 g a c c g g t a g c c g t g g t t g a g a c a c c c g a c c c g g c 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

1200 30 35 40 45 SKKFKVLGNTDRHSIKKNLI Cas9 SKKFKVLGNTDRHSIKKNLI ALFELHQAVGVPVADLLVQD 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 c 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 g

1260 50 55 60 65 GALLFDSGETAERTRLKRTA Cas9 GALLFDSGETAERTRLKRTA SGQQEVAAFCGLSGPQLSGG 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 a g a a c c c g g c t g a a g a g a a c c g c c 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 t c t t g g g c c g a c t t c t c t t g g c g g

BglII 1320 70 75 80 85 RRRYTRRKNRICYLQEIFSN Cas9 RRRYTRRKNRICYLQEIFSN SSSVGSPLVPDAIQLLDEAV 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 c 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 g

1380 90 95 100 105 EMAKVDDSFFHRLEESFLVE Cas9 EMAKVDDSFFHRLEESFLVE LHGLHVVAEEVSQFLGEQHF 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 a 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 t

1440 110 115 120 125 EDKKHERHPIFGNIVDEVAY Cas9 EDKKHERHPIFGNIVDEVAY LILLVLPVGDEAVDHVLHGV 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 c 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 g

1500 130 135 140 145 HEKYPTIYHLRKKLVDSTDK Cas9 HEKYPTIYHLRKKLVDSTDK VLLVGGDVVQSLFQHVAGVL 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 c

1560 150 155 160 165 ADLRLIYLALAHMIKFRGHF Cas9 ADLRLIYLALAHMIKFRGHF GVQPQDIQGQGVHDLEPAVE 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 g

1620 170 175 180 185 LIEGDLNPDNSDVDKLFIQL Cas9 LIEGDLNPDNSDVDKLFIQL QDLAVQVGVVAVHVLQEDLQ 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 c

1680 190 195 200 205 VQTYNQLFEENPINASGVDA Cas9 VQTYNQLFEENPINASGVDA HLGVVLQELFVGDVGAAHVG 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 g

1740 210 215 220 225 KAILSARLSKSRRLENLIAQ Cas9 KAILSARLSKSRRLENLIAQ LGDQRGSQALASPQFIQDGL 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 c

1800 230 235 240 245 LPGEKKNGLFGNLIALSLGL Cas9 LPGEKKNGLFGNLIALSLGL QGALLLIAQESVQNGQAQAQ 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 g 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 c

1860 250 255 260 265 TPNFKSNFDLAEDAKLQLSK Cas9 TPNFKSNFDLAEDAKLQLSK GGVELAVEVQGLIGFQLQAL 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 g 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 c

1920 270 275 280 285 DTYDDDLDNLLAQIGDQYAD Cas9 DTYDDDLDNLLAQIGDQYAD VGVVVVQVVQQGLDAVLVGV 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 g

1980 290 295 300 305 LFLAAKNLSDAILLSDILRV Cas9 LFLAAKNLSDAILLSDILRV QKQGGLVQGVGDQQAVDQSH 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 c

2040 310 315 320 325 NTEITKAPLSASMIKRYDEH Cas9 NTEITKAPLSASMIKRYDEH VGLDGLGGQAGRHDLSVVLV 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 g

2100 330 335 340 345 HQDLTLLKALVRQQLPEKYK Cas9 HQDLTLLKALVRQQLPEKYK VLVQGQQFSEHPLLQRLLVF 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 a 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 t

2160 350 355 360 365 EIFFDQSKNGYAGYIDGGAS Cas9 EIFFDQSKNGYAGYIDGGAS LNEEVLALVAVGAVNVASGA 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 c 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 g

2220 370 375 380 385 QEEFYKFIKPILEKMDGTEE Cas9 QEEFYKFIKPILEKMDGTEE LFLEVLEDLGDQFLHVAGLF 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 a 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 t

2280 390 395 400 405 LLVKLNREDLLRKQRTFDNG Cas9 LLVKLNREDLLRKQRTFDNG QEHLQVSLVQQPLLPGEVVA 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 c 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 g

2340 410 415 420 425 SIPHQIHLGELHAILRRQED Cas9 SIPHQIHLGELHAILRRQED ADGVLDVQSLQVGNQPPLFI 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 t 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 a

2400 430 435 440 445 FYPFLKDNREKIEKILTFRI Cas9 FYPFLKDNREKIEKILTFRI KVWEQLVVPFLDLLDQGEAD 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 c 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 g

PspOMIApaI 2460 450 455 460 465 PYYVGPLARGNSRFAWMTRK Cas9 PYYVGPLARGNSRFAWMTRK GVVHARQGPSVASEGPHGSL 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 c

2520 470 475 480 485 SEETITPWNFEEVVDKGASA Cas9 SEETITPWNFEEVVDKGASA ALFGDGGPVELFHHVLASGG 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 g

2580 490 495 500 505 QSFIERMTNFDKNLPNEKVL Cas9 QSFIERMTNFDKNLPNEKVL LAEDLPHGVEILVQGVLLHQ 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 c

2640 510 515 520 525 PKHSLLYEYFTVYNELTKVK Cas9 PKHSLLYEYFTVYNELTKVK GLVAQQVLVEGHIVLQGFHF 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 a 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 t

2700 530 535 540 545 YVTEGMRKPAFLSGEQKKAI Cas9 YVTEGMRKPAFLSGEQKKAI VHGLSHSLGGEQAALLFLGD 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 c 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 g

2760 550 555 560 565 VDLLFKTNRKVTVKQLKEDY Cas9 VDLLFKTNRKVTVKQLKEDY HVQQELGVPFHGHLLQFLVV 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 c 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 g

2820 570 575 580 585 FKKIECFDSVEISGVEDRFN Cas9 FKKIECFDSVEISGVEDRFN ELFDLAEVGHFDGAHFIPEV 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 c 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 g

PasI 2880 590 595 600 605 ASLGTYHDLLKIIKDKDFLD Cas9 ASLGTYHDLLKIIKDKDFLD GGQACVVIQQFNDLVLVEQV 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 c 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 g

EcoRV 2940 610 615 620 625 NEENEDILEDIVLTLTLFED Cas9 NEENEDILEDIVLTLTLFED ILFVLVNQFIDHQGQCQKLV 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 c 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 g

3000 630 635 640 645 REMIEERLKTYAHLFDDKVM Cas9 REMIEERLKTYAHLFDDKVM *AWRNSSLTI SLHDLFPQFGIGVQEVVFHH 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 c

3060 650 655 660 665 KQLKRRRYTGWGRLSRKLIN Cas9 KQLKRRRYTGWGRLSRKLIN FCSFRRLYVPQPLSLRFSIL LLQLPPSVGAPAPQAPLQDV 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 g

3120 670 675 680 685 GIRDKQSGKTILDFLKSDGF Cas9 GIRDKQSGKTILDFLKSDGF PMRSLCDPLVIRSKRFDSPK ADPVLLGALCDQIEQLGVAE 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 c 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 g

3180 690 695 700 705 ANRNFMQLIHDDSLTFKEDI Cas9 ANRNFMQLIHDDSLTFKEDI ALLFKMCSIWSSLRVKLSSM GVSVEHLQDVVVAQGKFLVD 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 c 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 g

3240 710 715 720 725 QKAQVSGQGDSLHEHIANLA Cas9 QKAQVSGQGDSLHEHIANLA WFAWTDPWPSLRCSCMALRA LFGLHGALAIAQVLVNGIQG 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 c 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 g

3300 730 735 740 745 GSPAIKKGILQTVKVVDELV Cas9 GSPAIKKGILQTVKVVDELV PLGAMLFPMRCVTFTTSSST AAGGNLLADQLCHLHHVLEH 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 c

3360 750 755 760 765 KVMGRHKPENIVIEMARENQ Cas9 KVMGRHKPENIVIEMARENQ FTIPRCLGSFMTISIALSFW FHHAPVLGLVDHDFHGSLVL 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 g 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 c

3420 770 775 780 785 TTQKGQKNSRERMKRIEEGI Cas9 TTQKGQKNSRERMKRIEEGI VVWFPCFFLRSLIFRISSPM GGLLSLLVAALSHLPDFLAD 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 g

3480 790 795 800 805 KELGSQILKEHPVENTQLQN Cas9 KELGSQILKEHPVENTQLQN FLQAALDQFFVGHFVGLQLV 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 c 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 g

3540 810 815 820 825 EKLYLYYLQNGRDMYVDQEL Cas9 EKLYLYYLQNGRDMYVDQEL LLQVQVVQLIPPIHVHVLFQ 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 c

3600 830 835 840 845 DINRLSDYDVDHIVPQSFLK Cas9 DINRLSDYDVDHIVPQSFLK VDVPQGVVIHVM 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 g 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 c

3660 850 855 860 865 DDSIDNKVLTRSDKNRGKSD Cas9 DDSIDNKVLTRSDKNRGKSD 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 g

3720 870 875 880 885 NVPSEEVVKKMKNYWRQLLN Cas9 NVPSEEVVKKMKNYWRQLLN 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 c 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 g

3780 890 895 900 905 AKLITQRKFDNLTKAERGGL Cas9 AKLITQRKFDNLTKAERGGL 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 c

3840 910 915 920 925 SELDKAGFIKRQLVETRQIT Cas9 SELDKAGFIKRQLVETRQIT 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 a 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 t

PmlI 3900 930 935 940 945 KHVAQILDSRMNTKYDENDK Cas9 KHVAQILDSRMNTKYDENDK 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 g 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 c

3960 950 955 960 965 LIREVKVITLKSKLVSDFRK Cas9 LIREVKVITLKSKLVSDFRK 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 g 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 c

4020 970 975 980 985 DFQFYKVREINNYHHAHDAY Cas9 DFQFYKVREINNYHHAHDAY 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 g

4080 990 995 1000 1005 LNAVVGTALIKKYPKLESEF Cas9 LNAVVGTALIKKYPKLESEF 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 c 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 g

4140 1010 1015 1020 1025 VYGDYKVYDVRKMIAKSEQE Cas9 VYGDYKVYDVRKMIAKSEQE 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 a 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 t

4200 1030 1035 1040 1045 IGKATAKYFFYSNIMNFFKT Cas9 IGKATAKYFFYSNIMNFFKT 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 c 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 g

4260 1050 1055 1060 1065 EITLANGEIRKRPLIETNGE Cas9 EITLANGEIRKRPLIETNGE 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 a 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 t

4320 1070 1075 1080 1085 TGEIVWDKGRDFATVRKVLS Cas9 TGEIVWDKGRDFATVRKVLS 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 c 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 g

4380 1090 1095 1100 1105 MPQVNIVKKTEVQTGGFSKE Cas9 MPQVNIVKKTEVQTGGFSKE 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 c

KflI 4440 1110 1115 1120 1125 SIRPKRNSDKLIARKKDWDP Cas9 SIRPKRNSDKLIARKKDWDP t c t a t c a g a 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 t a g a t a g t c t 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 a

4500 1130 1135 1140 1145 KKYGGFLWPTVAYSVLVVAK Cas9 KKYGGFLWPTVAYSVLVVAK a a g a a g t a c g g c g g c t t c c t g t g g 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 g a c a c c 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 t

BsaBI* 4560 1150 1155 1160 1165 VEKGKSKKLKSVKELLGITI Cas9 VEKGKSKKLKSVKELLGITI 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 c 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 g

4620 1170 1175 1180 1185 MERSSFEKNPIDFLEAKGYK Cas9 MERSSFEKNPIDFLEAKGYK 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 a 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 t

4680 1190 1195 1200 1205 EVKKDLIIKLPKYSLFELEN Cas9 EVKKDLIIKLPKYSLFELEN 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 c 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 g

BsmI 4740 1210 1215 1220 1225 GRKRMLASAKQLQKGNELAL Cas9 GRKRMLASAKQLQKGNELAL 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 a a g c a g 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 t t c g t c g a c g t c t t c c c t t t g c t t g a c c g g g a c

4800 1230 1235 1240 1245 PSKYVNFLYLASHYEKLKGS Cas9 PSKYVNFLYLASHYEKLKGS 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 g

4860 1250 1255 1260 1265 PEDNEQKQLFVEQHKHYLDE Cas9 PEDNEQKQLFVEQHKHYLDE 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 c

4920 1270 1275 1280 1285 IIEQISEFSKRVILADANLD Cas9 IIEQISEFSKRVILADANLD 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 g

4980 1290 1295 1300 1305 KVLSAYNKHRDKPIREQAEN Cas9 KVLSAYNKHRDKPIREQAEN 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 t 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 a

5040 1310 1315 1320 1325 IIHLFTLTRLGAPRAFKYFD Cas9 IIHLFTLTRLGAPRAFKYFD 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 g a c t g g g a g c c c c t a g a g c c t t c a a g t a c t t t g a c 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 c t g a c c c t c g g g g a t c t c g g a a g t t c a t g a a a c t g

5100 1330 1335 1340 1345 TTIDPKQYRSTKEVLDATLI Cas9 TTIDPKQYRSTKEVLDATLI a c c a c c a t c g a c c c c a a g c a g t a c a g a 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 c t g g t g g t a g c t g g g g t t c g t c a t g t c t 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 g

5160 1350 1355 1360 1365 HQSITGLYETRIDLSQLGGD Cas9 HQSITGLYETRIDLSQLGGD 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 t g a c 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 a c t g

EcoRI 5220 (in frame with Cas9) SGGSKRTADGSEFE 130 PKKKRK SV40 NLS SGGSKRTADGSEFEPKKKRK t c t g g c g g c t c a a a a a g a a c c g c c g a c g g c a g c g a a t t c g a g c c c a a g a a g a a g a g g a a a a g a c c g c c g a g t t t t t c t t g g c g g c t g c c g t c g c t t a a g c t c g g g t t c t t c t t c t c c t t t

BsmBIEsp3I 5280 V GSG 1 5 10 15 ATNFSLLKQAGDVEEN SV40 NLS P2A VGSGATNFSLLKQAGDVEEN g t c g g a a g c g g a g c t a c t a a c t t c a g c c t g c t g a a g c a g g c t g g a g a c g t g g a g g a g a a c c a g c c t t c g c c t c g a t g a t t g a a g t c g g a c g a c t t c g t c c g a c c t c t g c a c c t c c t c t t g

5340 PGP 1 M 1a V 5 10 15 SKGEELFTGVVPILV P2A EGFP 1 M 1a V 5 10 15 SKGEELFTGVVPILV EGFP PGPMVSKGEELFTGVVPILV c c t g g a c c t a t g g t g a g c a a g g g c g a g g a g c t g t t c a c c g g g g t g g t g c c c a t c c t g g t c g g a c c t g g a t a c c a c t c g t t c c c g c t c c t c g a c a a g t g g c c c c a c c a c g g g t a g g a c c a g

5400 20 25 30 35 ELDGDVNGHKFSVSGEGEGD EGFP 20 25 30 35 ELDGDVNGHKFSVSGEGEGD EGFP ELDGDVNGHKFSVSGEGEGD g a g c t g g a c g g c g a c g t a a a c g g c c a c a a g t t c a g c g t g t c c g g c g a g g g c g a g g g c g a t c t c g a c c t g c c g c t g c a t t t g c c g g t g t t c a a g t c g c a c a g g c c g c t c c c g c t c c c g c t a

5460 40 45 50 55 ATYGKLTLKFICTTGKLPVP EGFP 40 45 50 55 ATYGKLTLKFICTTGKLPVP EGFP ATYGKLTLKFICTTGKLPVP g c c a c c t a c g g c a a g c t g a c c c t g a a g t t c a t c t g c a c c a c c g g c a a g c t g c c c g t g c c c c g g t g g a t g c c g t t c g a c t g g g a c t t c a a g t a g a c g t g g t g g c c g t t c g a c g g g c a c g g g

5520 60 65 70 75 WPTLVTTLTYGVQCFSRYPD EGFP 60 65 70 75 WPTLVTTLTYGVQCFSRYPD EGFP WPTLVTTLTYGVQCFSRYPD 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 t g g a g t g c a g t g c t t c a g c c g c t a c c c c g a c 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 a c c t c a c g t c a c g a a g t c g g c g a t g g g g c t g

5580 80 85 90 95 HMKQHDFFKSAMPEGYVQER EGFP 80 85 90 95 HMKQHDFFKSAMPEGYVQER EGFP HMKQHDFFKSAMPEGYVQER c a c a t g a a g c a g c a c g a c t t c t t c a a g t c c g c c a t g c c c g a a g g c t a c g t c c a g g a g c g c g t g t a c t t c g t c g t g c t g a a g a a g t t c a g g c g g t a c g g g c t t c c g a t g c a g g t c c t c g c g

5640 100 105 110 115 TIFFKDDGNYKTRAEVKFEG EGFP 100 105 110 115 TIFFKDDGNYKTRAEVKFEG EGFP C T G C T G C C G T T G A T G T T C T G EXFP-R TIFFKDDGNYKTRAEVKFEG a c c a t c t t c t t c a a g g a c g a c g g c a a c t a c a a g a c c c g c g c c g a g g t g a a g t t c g a g g g c t g g t a g a a g a a g t t c c t g c t g c c g t t g a t g t t c t g g g c g c g g c t c c a c t t c a a g c t c c c g

5700 120 125 130 135 DTLVNRIELKGIDFKEDGNI EGFP 120 125 130 135 DTLVNRIELKGIDFKEDGNI EGFP DTLVNRIELKGIDFKEDGNI g a c a c c c t g g t g a a c c g c a t c g a g c t g a a g g g c a t c g a c t t c a a g g a g g a c g g c a a c a t c c t g t g g g a c c a c t t g g c g t a g c t c g a c t t c c c g t a g c t g a a g t t c c t c c t g c c g t t g t a g

5760 140 145 150 155 LGHKLEYNYNSHNVYIMADK EGFP 140 145 150 155 LGHKLEYNYNSHNVYIMADK EGFP LGHKLEYNYNSHNVYIMADK c t g g g g c a c a a g c t g g a g t a c a a c t a c a a c a g c c a c a a c g t c t a t a t c a t g g c c g a c a a g g a c c c c g t g t t c g a c c t c a t g t t g a t g t t g t c g g t g t t g c a g a t a t a g t a c c g g c t g t t c

5820 160 165 170 175 QKNGIKVNFKIRHNIEDGSV EGFP 160 165 170 175 QKNGIKVNFKIRHNIEDGSV EGFP QKNGIKVNFKIRHNIEDGSV c a g a a g a a c g g c a t c a a g g t g a a c t t c a a g a t c c g c c a c a a c a t c g a g g a c g g c a g c g t g g t c t t c t t g c c g t a g t t c c a c t t g a a g t t c t a g g c g g t g t t g t a g c t c c t g c c g t c g c a c

5880 180 185 190 195 QLADHYQQNTPIGDGPVLLP EGFP 180 185 190 195 QLADHYQQNTPIGDGPVLLP EGFP QLADHYQQNTPIGDGPVLLP c a g c t c g c c g a c c a c t a c c a g c a g a a c a c c c c c a t c g g c g a c g g c c c c g t g c t g c t g c c c g t c g a g c g g c t g g t g a t g g t c g t c t t g t g g g g g t a g c c g c t g c c g g g g c a c g a c g a c g g g

5940 200 205 210 215 DNHYLSTQSALSKDPNEKRD EGFP 200 205 210 215 DNHYLSTQSALSKDPNEKRD EGFP DNHYLSTQSALSKDPNEKRD g a c a a c c a c t a c c t g a g c a c c c a g t c c g c c c t g a g c a a a g a c c c c a a c g a g a a g c g c g a t c t g t t g g t g a t g g a c t c g t g g g t c a g g c g g g a c t c g t t t c t g g g g t t g c t c t t c g c g c t a

BsrGI 6000 220 225 230 235 HMVLLEFVTAAGITLGMDEL EGFP 220 225 230 235 HMVLLEFVTAAGITLGMDEL 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 HMVLLEFVTAAGITLGMDEL c a c a t g g t c c t g c t g g a g t t c g t g a c c g c c g c c g g g a t c a c t c t c g g c a t g g a c g a g c t g g t g t a c c a g g a c g a c c t c a a g c a c t g g c g g c g g c c c t a g t g a g a g c c g t a c c t g c t c g a c

AgeI 6060 YK SGGS 130 PKKKRKV * 1 HHHH EGFP SV40 NLS 6xHis YK (in frame with EGFP) SGGSPKKKRKV* EGFP YKSGGSPKKKRKV* t a c a a g t c t g g t g g t t c t c c c a a g a a g a a g a g g a a a g t c t a a c c g g t c a t c a t c a c c a t c a t g t t c a g a c c a c c a a g a g g g t t c t t c t t c t c c t t t c a g a t t g g c c a g t a g t a g t g g t a g

PmeI 6120 5 HH * 6xHis 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 c c a t t g a 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 g t g g t a a c t 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 a c

6180 bGH poly(A) signal 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 t t 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 a a

6240 bGH poly(A) signal 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 g g g g a t t a t t t t a c t c c t t t a a c g t a g c g t a a c a g a c t c a t c c a c a g t a a g a t a a g a c c c c c

BbsI 6300 bGH poly(A) signal 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 g g 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 c c

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

6420 M13 rev lac operator 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 M13/pUC Reverse c c t c t a g c t a g a g c t t g g c g t a a t 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 g g a g a t c g a t c t c g a a c c g c a t t a 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

6480 -10 -35 lac operator lac promoter G C G A M13/pUC Reverse 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 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 a g g g t g c c t g c g a g t g t t a a g g t g t g t t g t a t g 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 t c c c a c g g a

6540 CAP binding site a a t g a g t g a g c t a a c t c a c a t t a a 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 t t a c t c a c t c g a t t g a g t g t a a t 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

6600 a c c t g t c g t g c c a g c t g c a t t a a t 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 g g a c a g c a c g g t c g a c g t a a t t a 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

BspQISapI 6660 G A G C G A G T G A C T G A G C G A L4440 t t g g g c g c t c t t c c g c t t c c t c g c 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 a a c c c g c g a g a a g g c g a a g g a g c 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

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

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

DrdI 6840 ori t g c t g g c g t t t t t c c a t a g g c t c c 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 a c g a c c g c a a a a a g g t a t c c g a g 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

6900 ori T T T C T A T G G T C C G C A A A G G G pBR322ori-F g t c a g a g g t g g c g a a a c c c g a c a g 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 a g t c t c c a c c g c t t t g g g c t g t c 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

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

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

7080 ori 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 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 a g c a a g c g a g g t t c g a c c c g a c a c 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

7140 ori t a t c c g g t a a c t a t c g t c t t g a g t 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 a t a g g c c a t t g a t a g c a g a a c t 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

7200 ori 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 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 g t c g g t g a c c a t t g t c c t a a t c g t 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

7260 ori 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 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 t c a c c a c c g g a t t g a t g c c g a t g t 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

7320 ori a g c c a g t t a c c t t c g g a a a a a g a g 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 t c g g t c a a t g g a a g c c t t t t t c t c 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

7380 ori 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 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 c a t c g c c a c c a a a a a a a c a a a c g t 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

7440 a a g a t c c t t t g a t c t t t t c t a c g g g g t c t g a c a 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 t t c t a g g a a a c t a g a a a a g a t g c c c c a g a c t g t 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

7500 g g a t t t t g g t c a t g a g a t t a t c a a 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 c c t a a a a c c a g t a c t c t a a t a g t t 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

7560 285 *WHK AmpR *WHK 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 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 c t t c a a a a t t t a g t t a g a t t t c a t 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

7620 280 275 270 265 ILSAGIEAIQRNREDMTAQS AmpR ILSAGIEAIQRNREDMTAQS t a a t c a g t g a g g c a c c t a t c t c a g 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 a t t a g t c a c t c c g t g g a t a g a g t c 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

7680 260 255 250 245 GTTYIVVIRSPKGDPGLAAI AmpR GTTYIVVIRSPKGDPGLAAI t c c c c g t c g t g t a g a t a a c t a c g a 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 a g g g g c a g c a c a t c t a t t g a t g c t 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

BsaI 7740 240 235 230 225 IGRSGREGAGSKDAIFWGAP AmpR IGRSGREGAGSKDAIFWGAP t g a t a c c g c g a g a c c c a c g c t c a c 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 a c t a t g g c g c t c t g g g t g c g a g t g 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

7800 220 215 210 205 LASRLLPGAVKDAEMWDILQ AmpR LASRLLPGAVKDAEMWDILQ g a a g g g c c g a g c g c a g a a g t g g t c 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 c t t c c c g g c t c g c g t c t t c a c c a g 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

FspI 7860 200 195 190 185 QRSALTLLEGTLLKRLTTAM AmpR QRSALTLLEGTLLKRLTTAM 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 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 c a a c g g c c c t t c g a t c t c a t t c a t 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

7920 180 175 170 165 AVPMTTDREDNPIAENLEPE AmpR AVPMTTDREDNPIAENLEPE t t g c t a c a g g c a t c g t g g t g t c a c 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 a a c g a t g t c c g t a g c a c c a c a g t g 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

7980 160 155 150 145 WRDLRTVHDGMNHLFATLEK AmpR WRDLRTVHDGMNHLFATLEK c c c a a c g a t c a a g g c g a g t t a c a t 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 g g g t t g c t a g t t c c g c t c a a t g t 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

TsoIPvuI 8040 140 135 130 125 PGGITTLLLNAATNDSMTIA AmpR PGGITTLLLNAATNDSMTIA t c g g t c c t c c g a t c g t t g t c a g a a 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 a g c c a g g a g g c t a g c a a c a g t c t t 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

8100 120 115 110 105 ASCLERVTMGDTLHKETVPS AmpR ASCLERVTMGDTLHKETVPS c a g c a c t g c a t a a t t c t c t t a c t g 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 g t c g t g a c g t a t t a a g a g a a t g a c 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

8160 100 95 90 85 YEVLDNQSYHIRRGLQEQGA AmpR YEVLDNQSYHIRRGLQEQGA 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 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 t c a t g a g t t g g t t c a g t a a g a c t 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

8220 80 75 70 65 DIRSLVAGCLLVKFTSMMPF AmpR A T A A T A C C G C G C C A C A T A G C Amp-R DIRSLVAGCLLVKFTSMMPF c g t c a a t a c g g g a t a a t a c c g c g c 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 g c a g t t a t g c c c t a t t a t g g c g c g 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

8280 60 55 50 45 REEPRFSELIKGSNLDLEIY AmpR REEPRFSELIKGSNLDLEIY a a c g t t c t t c g g g g c g a a a a c t c t 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 t t g c a a g a a g c c c c g c t t t t g a g a 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

8340 40 35 30 25 GVRAGLQDEADKVKVLTEPH AmpR GVRAGLQDEADKVKVLTEPH a a c c c a c t c g t g c a c c c a a c t g a t 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 t t g g g t g a g c a c g t g g g t t g a c t a 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

8400 20 15 10 5 signal sequence AFVPLCFAAFFPILAVRFHQ AmpR AFVPLCFAAFFPILAVRFHQ g a g c a a a a a c a g g a a g g c a a a a t g 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 c t c g t t t t t g t c c t t c c g t t t t a 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

SspI 8460 1 signal sequence ISM AmpR AmpR promoter ISM g a a t a c t c a t a c t c t t c c t t t t t c 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 c t t a t g a g t a t g a g a a g g a a a a a g 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

8520 AmpR promoter 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 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 a c t c g c c t a t g t a t a a a c t t a c a t 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

BspDI*ClaI*SgrDI 8580 t t c c c c g a a a a g t g c c a c c t g a c g t c g a c g g a t c g g g a g a t c g a t c t c c c g a t c c c c t a g a a g g g g c t t t t c a c g g t g g a c t g c a g c t g c c t a g c c c t c t a g c t a g a g g g c t a g g g g a t c

8640 A T G T T A G A C G A G A C T A C G G C pRS-marker g g t c g a c t c t c a g t a c a a t c t g c t c t g a t g c c g c a t a g t t a a g c c a g t a t c t g c t c c c t g c c a g c t g a g a g t c a t g t t a g a c g a g a c t a c g g c g t a t c a a t t c g g t c a t a g a c g a g g g a c

8700 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 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 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 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

NruIMfeI 8760 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 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 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 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

SpeIMluI 8820 CMV enhancer a t g t a c g g g c c a g a t a t a c g c g t t 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 a c a t g c c c g g t c t a t a t g c g c a a 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

8880 CMV enhancer 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 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 a t g c c c c a g t a a t c a a g t a t c g g 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

8940 CMV enhancer a t 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 a t g a c g t a t g 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 t a c t g c a t a c

9000 CMV enhancer t t c c c a t a g t a a c g c c a a t a 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 g g g t a t c a t t g c g g t t a t 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

3ʹ 5ʹ 9033 CMV enhancer a a a c t g c c c a c t t g g c a g t a c a t c a a g t g t a t c t t t g a c g g g t g a a c c g t c a t g t a g t t c a c a t a 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.