Addgene B-HypaR-SpCas9 Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: B-HypaR-SpCas9 Sequencing Result


Map View

SnaBI (7959) NdeI (7853) KpnI (7696) Acc65I (7692) pBR322ori-F (7527 .. 7546) SacII (6729) FspI (6571) PvuI (6425) Amp-R (6225 .. 6244) pBRforEco (5816 .. 5834) pGEX 3' (5756 .. 5778) pRS-marker (5637 .. 5656) PsiI (5523) F1ori-F (5462 .. 5483) F1ori-R (5252 .. 5271) PluTI (5095) SfoI (5093) NarI (5092) KasI (5091) SbfI (5088) NotI (4943) BGH-rev (4728 .. 4745) EcoRI (4704) FseI (4681) nucleoplasmin NLS BsmI (4189) BseRI (314) PaqCI (419) AgeI (441) BglII (820) BmgBI (1109) PspOMI (1925) ApaI (1929) PasI (2339) EcoRV (2423) PmlI (3359) AflIII - MluI (3526) KflI (3926) BsaBI * (4047) B-HypaR-SpCas9 8191 bp

Sequence View

5ʹ 3ʹ 60 XXGXXXGGGGGGGGGGGGGG g g b s g d g g g g g k g g k k n g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g s g g g g g v g g c c v s c h c c c c c m c c m m n c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c s c c c c c b c c

120 XGXGGXGGGAGRGGEVRRQP g g s g g g g s g g g g g g g g g s g g g g g g c g g g g c g g g g c g a g g c g g a g a g g t g c g g c g g c a g c c c c s c c c c s c c c c c c c c c s c c c c c c g c c c c g c c c c g c t c c g c c t c t c c a c g c c g c c g t c g g

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

240 hybrid intron IKSEARGGRESLRAAFAPCP t a t a a a a a g c g a a g c g c g c g g c g g g c g g g a g t c g c t g c g c g c t g c c t t c g c c c c g t g c c c a t a t t t t t c g c t t c g c g c g c c g c c c g c c c t c a g c g a c g c g c g a c g g a a g c g g g g c a c g g g

300 hybrid intron APPPPRAARPGSD* c g c t c c g c c g c c g c c t c g c g c c g c c c g c c c c g g c t c t g a c t g a c c g c g t t a c t c c c a c a g g c g a g g c g g c g g c g g a g c g c g g c g g g c g g g g c c g a g a c t g a c t g g c g c a a t g a g g g t g t c

BseRI 360 hybrid intron g t g a g c g g g c g g g a c g g c c c t t c t c c t c c g g g c t g t a a t t a g c t g a g c a a g a g g t a a g g g c a c t c g c c c g c c c t g c c g g g a a g a g g a g g c c c g a c a t t a a t c g a c t c g t t c t c c a t t c c c

PaqCI 420 hybrid intron t t t a a g g g a t g g t t g g t t g g t g g g g t a t t a a t g t t t a a t t a c c t g g a g c a c c t g c c t g a a a a a t t c c c t a c c a a c c a a c c a c c c c a t a a t t a c a a a t t a a t g g a c c t c g t g g a c g g a c t t

AgeI 480 1 5 DYKDHDGD hybrid intron 3xFLAG Kozak sequence MDYKDHDGD *TPGTGGHVILVVVSV a t c a c t t t t t t t c a g g t t g g a c c g g t g c c a c c a t g g a c t a t a a g g a c c a c g a c g g a g a c t t a g t g a a a a a a a g t c c a a c c t g g c c a c g g t g g t a c c t g a t a t t c c t g g t g c t g c c t c t g a

540 10 15 20 YKDHDIDYKDDDDK MA PKKK 3xFLAG SV40 NLS YKDHDIDYKDDDDKMAPKKK VLIMINIVFVIVILHGWLLL a c a a g g a t c a t g a t a t t g a t t a c a a a g a c g a t g a c g a t a a g a t g g c c c c a a a g a a g a a g c t g t t c c t a g t a c t a t a a c t a a t g t t t c t g c t a c t g c t a t t c t a c c g g g g t t t c t t c t t c g

600 130 RKV (in frame with SV40 NLS) GIHGVPAA 5 10 DKKYSIGLD SV40 NLS Cas9 RKVGIHGVPAADKKYSIGLD PLDTDVSDWCGVLLVADAQV g g a a g g t c g g t a t c c a c g g a g t c c c a g c a g c c g a c a a g a a g t a c a g c a t c g g c c t g g a c a c c t t c c a g c c a t a g g t g c c t c a g g g t c g t c g g c t g t t c t t c a t g t c g t a g c c g g a c c t g t

660 15 20 25 30 IGTNSVGWAVITDEYKVPSK Cas9 IGTNSVGWAVITDEYKVPSK DAGVRHAPGHDGVLVLHGAL 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 a g c a a g a 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 t c g t t c t

720 35 40 45 50 KFKVLGNTDRHSIKKNLIGA Cas9 KFKVLGNTDRHSIKKNLIGA FELHQAVGVPVADLLVQDSG 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 g g a g c c c 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 c c t c g g g

780 55 60 65 70 LLFDSGETAEATRLKRTARR Cas9 LLFDSGETAEATRLKRTARR QQEVAAFCGLGGPQLSGGSS 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 c a g a a g a a 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 g t c t t c t t

BglII 840 75 80 85 90 RYTRRKNRICYLQEIFSNEM Cas9 RYTRRKNRICYLQEIFSNEM SVGSPLVPDAIQLLDEAVLH 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 g a g a t g g 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 c t c t a c c

900 95 100 105 110 AKVDDSFFHRLEESFLVEED Cas9 AKVDDSFFHRLEESFLVEED GLHVVAEEVSQFLGEQHFLI 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 g a g g a t a 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 c t c c t a t

960 115 120 125 130 KKHERHPIFGNIVDEVAYHE Cas9 KKHERHPIFGNIVDEVAYHE LLVLPVGDEAVDHVLHGVVL 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 a c g a g a 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 g t g c t c t

1020 135 140 145 150 KYPTIYHLRKKLVDSTDKAD Cas9 KYPTIYHLRKKLVDSTDKAD LVGGDVVQSLFQHVAGVLGV 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 c c g a c c 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 c g g c t g g

1080 155 160 165 170 LRLIYLALAHMIKFRGHFLI Cas9 LRLIYLALAHMIKFRGHFLI QPQDIQGQGVHDLEPAVEQD 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 t g a t c 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 g a c t a g c

BmgBI 1140 175 180 185 190 EGDLNPDNSDVDKLFIQLVQ Cas9 EGDLNPDNSDVDKLFIQLVQ LAVQVGVVAVHVLQEDLQHL 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 t g c a g a 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 c a c g t c t

1200 195 200 205 210 TYNQLFEENPINASGVDAKA Cas9 TYNQLFEENPINASGVDAKA GVVLQELFVGDVGAAHVGLG 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 a a g g c c a 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 t t c c g g t

1260 215 220 225 230 ILSARLSKSRRLENLIAQLP Cas9 ILSARLSKSRRLENLIAQLP DQRGSQALASPQFIQDGLQG 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 c t g c c c g 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 g a c g g g c

1320 235 240 245 250 GEKKNGLFGNLIALSLGLTP Cas9 GEKKNGLFGNLIALSLGLTP ALLLIAQESVQNGQAQAQGG 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 a c c c c c a 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 t g g g g g t

1380 255 260 265 270 NFKSNFDLAEDAKLQLSKDT Cas9 NFKSNFDLAEDAKLQLSKDT VELAVEVQGLIGFQLQALVG 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 g a c a c c 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 c t g t g g a

1440 275 280 285 290 YDDDLDNLLAQIGDQYADLF Cas9 YDDDLDNLLAQIGDQYADLF VVVVQVVQQGLDAVLVGVQK 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 t t c 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 g a c a a a g

1500 295 300 305 310 LAAKNLSDAILLSDILRVNT Cas9 LAAKNLSDAILLSDILRVNT QGGLVQGVGDQQAVDQSHVG 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 a a c a c c 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 t t g t g g c

1560 315 320 325 330 EITKAPLSASMIKRYDEHHQ Cas9 EITKAPLSASMIKRYDEHHQ LDGLGGQAGRHDLSVVLVVL 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 c a c c a g g 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 g t g g t c c

1620 335 340 345 350 DLTLLKALVRQQLPEKYKEI Cas9 DLTLLKALVRQQLPEKYKEI VQGQQFSEHPLLQRLLVFLN 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 a g a t t t 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 c t c t a a a

1680 355 360 365 370 FFDQSKNGYAGYIDGGASQE Cas9 FFDQSKNGYAGYIDGGASQE EEVLALVAVGAVNVASGALF 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 a g g a a g 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 g t c c t t c

1740 375 380 385 390 EFYKFIKPILEKMDGTEELL Cas9 EFYKFIKPILEKMDGTEELL LEVLEDLGDQFLHVAGLFQE 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 c t g c t c g 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 g a c g a g c

1800 395 400 405 410 VKLNREDLLRKQRTFDNGSI Cas9 VKLNREDLLRKQRTFDNGSI HLQVSLVQQPLLPGEVVAAD 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 a g c a t c 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 t c g t a g g

1860 415 420 425 430 PHQIHLGELHAILRRQEDFY Cas9 PHQIHLGELHAILRRQEDFY GVLDVQSLQVGNQPPLFIKV 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 t t t a c c 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 a a a a t g g

1920 435 440 445 450 PFLKDNREKIEKILTFRIPY Cas9 PFLKDNREKIEKILTFRIPY WEQLVVPFLDLLDQGEADGV 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 c c c t a c t 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 g g g a t g a

PspOMIApaI 1980 455 460 465 470 YVGPLARGNSRFAWMTRKSE Cas9 YVGPLARGNSRFAWMTRKSE VHARQGPSVASEGPHGSLAL 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 a g c g a g g 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 t c g c t c c

2040 475 480 485 490 ETITPWNFEEVVDKGASAQS Cas9 ETITPWNFEEVVDKGASAQS FGDGGPVELFHHVLASGGLA 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 c a g a g c t 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 g t c t c g a

2100 495 500 505 510 FIERMTNFDKNLPNEKVLPK Cas9 FIERMTNFDKNLPNEKVLPK EDLPHGVEILVQGVLLHQGL 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 c c c a a g c 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 g g g t t c g

2160 515 520 525 530 HSLLYEYFTVYNELTKVKYV Cas9 HSLLYEYFTVYNELTKVKYV VAQQVLVEGHIVLQGFHFVH 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 t a c g t g a 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 a t g c a c t

2220 535 540 545 550 TEGMRKPAFLSGEQKKAIVD Cas9 TEGMRKPAFLSGEQKKAIVD GLSHSLGGEQAALLFLGDHV 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 g t g g a c c 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 c a c c t g g

2280 555 560 565 570 LLFKTNRKVTVKQLKEDYFK Cas9 LLFKTNRKVTVKQLKEDYFK QQELGVPFHGHLLQFLVVEL 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 t t c a a g a 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 a a g t t c t

PasI 2340 575 580 585 590 KIECFDSVEISGVEDRFNAS Cas9 KIECFDSVEISGVEDRFNAS FDLAEVGHFDGAHFIPEVGG 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 g c c t c c c 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 c g g a g g g

2400 595 600 605 610 LGTYHDLLKIIKDKDFLDNE Cas9 LGTYHDLLKIIKDKDFLDNE QACVVIQQFNDLVLVEQVIL 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 a a t g a 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 t t a c t c c

EcoRV 2460 615 620 625 630 ENEDILEDIVLTLTLFEDRE Cas9 ENEDILEDIVLTLTLFEDRE FVLVNQFIDHQGQCQKLVSL 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 a g a g a g a 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 t c t c t c t

2520 635 640 645 650 MIEERLKTYAHLFDDKVMKQ Cas9 MIEERLKTYAHLFDDKVMKQ HDLFPQFGIGVQEVVFHHLL *AWRNSSLTIFCS 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 a a g c a g c 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 t t c g t c g

2580 655 660 665 670 LKRRRYTGWGALSRKLINGI Cas9 LKRRRYTGWGALSRKLINGI QLPPSVGAPARQAPLQDVAD FRRLYVPQPASLRFSILPMR 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 g c g c t g a g c c g g a a g c t g a t c a a c g g c a t c c 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 c g c g a c t c g g c c t t c g a c t a g t t g c c g t a g g

2640 675 680 685 690 RDKQSGKTILDFLKSDGFAN Cas9 RDKQSGKTILDFLKSDGFAN AVLLGALCDQIEQLGVAEGV SLCDPLVIRSKRFDSPKALL g c 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 g c c a a c a c g 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 c g g t t g t

2700 695 700 705 710 RAFAALIADDSLTFKEDIQK Cas9 RAFAALIADDSLTFKEDIQK SGKCGQDSVVAQGKFLVDLF AKAARIASSLRVKLSSMWFA g a g c c t t t g c a g c c c t g a t c g c t g a c g a c a g c c t g a c c t t t a a a g a g g a c a t c c a g a a a g c t c g g a a a c g t c g g g a c t a g c g a c t g c t g t c g g a c t g g a a a t t t c t c c t g t a g g t c t t t c

2760 715 720 725 730 AQVSGQGDSLHEHIANLAGS Cas9 AQVSGQGDSLHEHIANLAGS GLHGALAIAQVLVNGIQGAA WTDPWPSLRCSCMALRAPLG 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 g c a g c 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 c c g t c g g

2820 735 740 745 750 PAIKKGILQTVKVVDELVKV Cas9 PAIKKGILQTVKVVDELVKV GGNLLADQLCHLHHVLEHFH AMLFPMRCVTFTTSSSTFTI 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 a a a g t g a 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 t t t c a c t

2880 755 760 765 770 MGRHKPENIVIEMARENQTT Cas9 MGRHKPENIVIEMARENQTT HAPVLGLVDHDFHGSLVLGG PRCLGSFMTISIALSFWVVW 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 a c c a c c c 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 t g g t g g g

2940 775 780 785 790 QKGQKNSRERMKRIEEGIKE Cas9 QKGQKNSRERMKRIEEGIKE LLSLLVAALSHLPDFLADFL FPCFFLRSLIFRISSPM 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 a a a g a g c 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 t t t c t c g

3000 795 800 805 810 LGSQILKEHPVENTQLQNEK Cas9 LGSQILKEHPVENTQLQNEK QAALDQFFVGHFVGLQLVLL 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 g a g a a g c 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 c t c t t c g

3060 815 820 825 830 LYLYYLQNGRDMYVDQELDI Cas9 LYLYYLQNGRDMYVDQELDI QVQVVQLIPPIHVHVLFQVD 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 g a c a t c a 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 c t g t a g t

3120 835 840 845 850 NRLSDYDVDHIVPQSFLKDD Cas9 NRLSDYDVDHIVPQSFLKDD VPQGVVIHVM 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 g a c g a c 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 c t g c t g a

3180 855 860 865 870 SIDNKVLTRSDKNRGKSDNV Cas9 SIDNKVLTRSDKNRGKSDNV 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 a a c g t g c 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 t t g c a c g

3240 875 880 885 890 PSEEVVKKMKNYWRQLLNAK Cas9 PSEEVVKKMKNYWRQLLNAK 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 g c c a a g c 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 c g g t t c g

3300 895 900 905 910 LITQRKFDNLTKAERGGLSE Cas9 LITQRKFDNLTKAERGGLSE 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 a g c g a a c 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 t c g c t t g

PmlI 3360 915 920 925 930 LDKAGFIKRQLVETRQITKH Cas9 LDKAGFIKRQLVETRQITKH 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 a a g c a c 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 t t c g t g c

3420 935 940 945 950 VAQILDSRMNTKYDENDKLI Cas9 VAQILDSRMNTKYDENDKLI 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 c t g a t c 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 g a c t a g g

3480 955 960 965 970 REVKVITLKSKLVSDFRKDF Cas9 REVKVITLKSKLVSDFRKDF 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 t t t c c 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 c t a a a g g

AflIIIMluI 3540 975 980 985 990 QFYKVREINNYHHAHDAYLN Cas9 QFYKVREINNYHHAHDAYLN 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 g t a c c t g a a c 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 c a t g g a c t t g c

3600 995 1000 1005 1010 AVVGTALIKKYPKLGGKVYD Cas9 AVVGTALIKKYPKLGGKVYD 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 g t g g c a a g g t g t a c g a t g 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 c a c c g t t c c a c a t g c t a c

3660 1015 1020 1025 1030 VRKMIAKSEQEIGKATAKYF Cas9 VRKMIAKSEQEIGKATAKYF t a c g g a a g a t g a t c g c c a a g a g c g a g c a g g a a a t c g g c a a g g c t a c c g c c a a g t a c t t c t a t g c c t t c t a c t a g c g g t t c t c g c t c g t c c t t t a g c c g t t c c g a t g g c g g t t c a t g a a g a

3720 1035 1040 1045 1050 FYSNIMNFFKTEITLANGEI Cas9 FYSNIMNFFKTEITLANGEI 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 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 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 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

3780 1055 1060 1065 1070 RKRPLIETNGETGEIVWDKG Cas9 RKRPLIETNGETGEIVWDKG 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 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 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 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

3840 1075 1080 1085 1090 RDFATVRKVLSMPQVNIVKK Cas9 RDFATVRKVLSMPQVNIVKK 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 a t g c c c c a a g t g a a t a t c g t g a a a a a g a c c c t a a a a c g g t g g c a c g c c t t t c a c g a c t c g t a c g g g g t t c a c t t a t a g c a c t t t t t c t

3900 1095 1100 1105 1110 TEVQTGGFSKESILPKRNSD Cas9 TEVQTGGFSKESILPKRNSD 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 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 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 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

KflI 3960 1115 1120 1125 1130 KLIARKKDWDPKKYGGFDSP Cas9 KLIARKKDWDPKKYGGFDSP 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 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 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 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

4020 1135 1140 1145 1150 TVAYSVLVVAKVEKGKSKKL Cas9 TVAYSVLVVAKVEKGKSKKL 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 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 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 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

BsaBI* 4080 1155 1160 1165 1170 KSVKELLGITIMERSSFEKN Cas9 KSVKELLGITIMERSSFEKN 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 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 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 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

4140 1175 1180 1185 1190 PIDFLEAKGYKEVKKDLIIK Cas9 PIDFLEAKGYKEVKKDLIIK 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 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 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 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

BsmI 4200 1195 1200 1205 1210 LPKYSLFELENGRKRMLASA Cas9 LPKYSLFELENGRKRMLASA 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 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 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 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

4260 1215 1220 1225 1230 GELQKGNELALPSKYVNFLY Cas9 GELQKGNELALPSKYVNFLY 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 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 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 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

4320 1235 1240 1245 1250 LASHYEKLKGSPEDNEQKQL Cas9 LASHYEKLKGSPEDNEQKQL 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 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 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 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

4380 1255 1260 1265 1270 FVEQHKHYLDEIIEQISEFS Cas9 FVEQHKHYLDEIIEQISEFS 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 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 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 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

4440 1275 1280 1285 1290 KRVILADANLDKVLSAYNKH Cas9 KRVILADANLDKVLSAYNKH 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 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 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 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

4500 1295 1300 1305 1310 RDKPIREQAENIIHLFTLTN Cas9 RDKPIREQAENIIHLFTLTN 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 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 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 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

4560 1315 1320 1325 1330 LGAPAAFKYFDTTIDRKRYT Cas9 LGAPAAFKYFDTTIDRKRYT 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 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 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 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

4620 1335 1340 1345 1350 STKEVLDATLIHQSITGLYE Cas9 STKEVLDATLIHQSITGLYE 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 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 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 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

4680 1355 1360 TRIDLSQLGGD 1 5 KRPAATKKA Cas9 nucleoplasmin NLS TRIDLSQLGGDKRPAATKKA 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 c a a a a g g c c g g c g g c c a c g a a a a a g g c c g g 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 g t t t t c c g g c c g c c g g t g c t t t t t c c g g c

EcoRIFseI 4740 10 15 GQAKKKK * nucleoplasmin NLS bGH poly(A) signal G G A G C T G A C A C G G BGH-rev GQAKKKK* g c c a g g c a a a a a a g a a a a a g t a a g a a t t c c t a g a g c t c g c t g a t c a g c c t c g a c t g t g c c c g g t c c g t t t t t t c t t t t t c a t t c t t a a g g a t c t c g a g c g a c t a g t c g g a g c t g a c a c g g

4800 bGH poly(A) signal A A G A T BGH-rev t t c t a g t t g c c a g c c a t c t g t t g t t t g c c c c t c c c c c g t g c c t t c c t t g a c c c t g g a a g g a a g a t c a a c g g t c g g t a g a c a a c a a a c g g g g a g g g g g c a c g g a a g g a a c t g g g a c c t t c c

4860 bGH poly(A) signal t g c c a c t c c c a c t g t c c t t t c c t a a t a a a a t g a g g a a a t t g c a t c g c a t t g t c t g a g t a g a c g g t g a g g g t g a c a g g a a a g g a t t a t t t t a c t c c t t t a a c g t a g c g t a a c a g a c t c a t c

4920 bGH poly(A) signal g t g t c a t t c t a t t c t g g g g g g t g g g g t g g g g c a g g a c a g c a a g g g g g a g g a t t g g g a a g a c a c a g t a a g a t a a g a c c c c c c a c c c c a c c c c g t c c t g t c g t t c c c c c t c c t a a c c c t t c t

NotI 4980 bGH poly(A) signal AAV2 ITR AAV2 ITR g a a t a g c a g g c a t g c t g g g g a g c g g c c g c a g g a a c c c c t a g t g a t g g a g t t g g c c a c t c c c t t a t c g t c c g t a c g a c c c c t c g c c g g c g t c c t t g g g g a t c a c t a c c t c a a c c g g t g a g g

5040 AAV2 ITR AAV2 ITR c t c t c t g c g c g c t c g c t c g c t c a c t g a g g c c g g g c g a c c a a a g g t c g c c c g a c g c c c g g g g a g a g a c g c g c g a g c g a g c g a g t g a c t c c g g c c c g c t g g t t t c c a g c g g g c t g c g g g c c c

SbfIKasINarISfoIPluTI 5100 AAV2 ITR AAV2 ITR c t t t g c c c g g g c g g c c t c a g t g a g c g a g c g a g c g c g c a g c t g c c t g c a g g g g c g c c t g a t g a a a c g g g c c c g c c g g a g t c a c t c g c t c g c t c g c g c g t c g a c g g a c g t c c c c g c g g a c t a

5160 g c g g t a t t t t c t c c t t a c g c a t c t g t g c g g t a t t t c a c a c c g c a t a c g t c a a a g c a a c c a c g c c a t a a a a g a g g a a t g c g t a g a c a c g c c a t a a a g t g t g g c g t a t g c a g t t t c g t t g g t

5220 f1 ori t a g t a c g c g c c c t g t a g c g g c g c a t t a a g c g c g g c g g g t g t g g t g g t t a c g c g c a g c g t g a t c a t g c g c g g g a c a t c g c c g c g t a a t t c g c g c c g c c c a c a c c a c c a a t g c g c g t c g c a c

5280 f1 ori G A G G A A A G C G A A A G A A G G G A F1ori-R a c c g c t a c a c t t g c c a g c g c c t t a g c g c c c g c t c c t t t c g c t t t c t t c c c t t c c t t t c t c t g g c g a t g t g a a c g g t c g c g g a a t c g c g g g c g a g g a a a g c g a a a g a a g g g a a g g a a a g a g

5340 f1 ori 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 a 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 t

5400 f1 ori t t t a g t g c t t t a c g g c a c c t c g a c c c c a a a a a a c t t g a t t t g g g t g a t g g t t c a c g t a g t a a a t c a c g a a a t g c c g t g g a g c t g g g g t t t t t t g a a c t a a a c c c a c t a c c a a g t g c a t c a

5460 f1 ori 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 t 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 a

5520 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 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 t c t a t c t c g g g c t a t t c t t t t g a t t 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 a g a t a g a g c c c g a t a a g a a a a c t a

PsiI 5580 f1 ori 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 t c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a a a a 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 a g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t t t t t

5640 f1 ori A T G T pRS-marker t t t a a c g c g a a t t t t a a c a a a a t a t t a a c g t t t a c a a t t t t a t g g t g c a c t c t c a g t a c a a a a t t g c g c t t a a a a t t g t t t t a t a a t t g c a a a t g t t a a a a t a c c a c g t g a g a g t c a t g t

5700 T A G A C G A G A C T A C G G C pRS-marker a t c t g c t c t g a t g c c g c a t a g t t a a g c c a g c c c c g a c a c c c g c c a a c a c c c g c t g a c g c g t a g a c g a g a c t a c g g c g t a t c a a t t c g g t c g g g g c t g t g g g c g g t t g t g g g c g a c t g c g c

5760 C C G G G pGEX 3' c c c t g a c g g g c t t g t c t g c t c c c g g c a t c c g c t t a c a g a c a a g c t g t g a c c g t c t c c g g g g g g a c t g c c c g a a c a g a c g a g g g c c g t a g g c g a a t g t c t g t t c g a c a c t g g c a g a g g c c c

5820 A G C T G C A T G T G T C A G A G G pGEX 3' C G G A G pBRforEco a g c t g c a t g t g t c a g a g g t t t t c a c c g t c a t c a c c g a a a c g c g c g a g a c g a a a g g g c c t c t c g a c g t a c a c a g t c t c c a a a a g t g g c a g t a g t g g c t t t g c g c g c t c t g c t t t c c c g g a g

5880 C A C T A T G C G G A T A A pBRforEco g t g a t a c g c c t a t t t t t a t a g g t t a a t g t c a t g a t a a t a a t g g t t t c t t a g a c g t c a g g t c a c t a t g c g g a t a a a a a t a t c c a a t t a c a g t a c t a t t a t t a c c a a a g a a t c t g c a g t c c a

5940 AmpR promoter g g c a c t t t t c g g g g a a a t g t g c g c g g a a c c c c t a t t t g t t t a t t t t t c t a a a t a c a t t c a c c g t g a a a a g c c c c t t t a c a c g c g c c t t g g g g a t a a a c a a a t a a a a a g a t t t a t g t a a g t

6000 AmpR promoter a a t a t g t a t c c g c t c a t g a g a c a a t a a c c c t g a t a a a t g c t t c a a t a a t a t t g a a a a a g g t t a t a c a t a g g c g a g t a c t c t g t t a t t g g g a c t a t t t a c g a a g t t a t t a t a a c t t t t t c c

6060 1 5 10 15 signal sequence MSIQHFRVALIPFFAAFC AmpR promoter AmpR MSIQHFRVALIPFFAAFC a a g a g t a t g a g t a t t c a a c a t t t c c g t g t c g c c c t t a t t c c c t t t t t t g c g g c a t t t t g c t t c t c a t a c t c a t a a g t t g t a a a g g c a c a g c g g g a a t a a g g g a a a a a a c g c c g t a a a a c g

6120 20 signal sequence LPVFA 25 30 35 HPETLVKVKDAEDQL AmpR LPVFAHPETLVKVKDAEDQL c t t c c t g t t t t t g c t c a c c c a g a a a c g c t g g t g a a a g t a a a a g a t g c t g a a g a t c a g t t g g a a g g a c a a a a a c g a g t g g g t c t t t g c g a c c a c t t t c a t t t t c t a c g a c t t c t a g t c a a c

6180 40 45 50 55 GARVGYIELDLNSGKILESF AmpR GARVGYIELDLNSGKILESF g g t g c a c g a g t g g g t t a c a t c g a a c t g g a t c t c a a c a g c g g t a a g a t c c t t g a g a g t t t t c c a c g t g c t c a c c c a a t g t a g c t t g a c c t a g a g t t g t c g c c a t t c t a g g a a c t c t c a a a a

6240 60 65 70 75 RPEERFPMMSTFKVLLCGAV AmpR C G A T A C A C C G C G C C A T Amp-R RPEERFPMMSTFKVLLCGAV c g c c c c g a a g a a c g t t t t c c a a t g a t g a g c a c t t t t a a a g t t c t g c t a t g t g g c g c g g t a g c g g g g c t t c t t g c a a a a g g t t a c t a c t c g t g a a a a t t t c a a g a c g a t a c a c c g c g c c a t

6300 80 85 90 95 LSRIDAGQEQLGRRIHYSQN AmpR A A T A Amp-R LSRIDAGQEQLGRRIHYSQN t t a t c c c g t a t t g a c g c c g g g c a a g a g c a a c t c g g t c g c c g c a t a c a c t a t t c t c a g a a t a a t a g g g c a t a a c t g c g g c c c g t t c t c g t t g a g c c a g c g g c g t a t g t g a t a a g a g t c t t a

6360 100 105 110 115 DLVEYSPVTEKHLTDGMTVR AmpR DLVEYSPVTEKHLTDGMTVR g a c t t g g t t g a g t a c t c a c c a g t c a c a g a a a a g c a t c t t a c g g a t g g c a t g a c a g t a a g a c t g a a c c a a c t c a t g a g t g g t c a g t g t c t t t t c g t a g a a t g c c t a c c g t a c t g t c a t t c t

6420 120 125 130 135 ELCSAAITMSDNTAANLLLT AmpR ELCSAAITMSDNTAANLLLT g a a t t a t g c a g t g c t g c c a t a a c c a t g a g t g a t a a c a c t g c g g c c a a c t t a c t t c t g a c a c t t a a t a c g t c a c g a c g g t a t t g g t a c t c a c t a t t g t g a c g c c g g t t g a a t g a a g a c t g t

PvuI 6480 140 145 150 155 TIGGPKELTAFLHNMGDHVT AmpR TIGGPKELTAFLHNMGDHVT a c g a t c g g a g g a c c g a a g g a g c t a a c c g c t t t t t t g c a c a a c a t g g g g g a t c a t g t a a c t t g c t a g c c t c c t g g c t t c c t c g a t t g g c g a a a a a a c g t g t t g t a c c c c c t a g t a c a t t g a

6540 160 165 170 175 RLDRWEPELNEAIPNDERDT AmpR RLDRWEPELNEAIPNDERDT c g c c t t g a t c g t t g g g a a c c g g a g c t g a a t g a a g c c a t a c c a a a c g a c g a g c g t g a c a c c g c g g a a c t a g c a a c c c t t g g c c t c g a c t t a c t t c g g t a t g g t t t g c t g c t c g c a c t g t g g

FspI 6600 180 185 190 195 TMPVAMATTLRKLLTGELLT AmpR TMPVAMATTLRKLLTGELLT a c g a t g c c t g t a g c a a t g g c a a c a a c g t t g c g c a a a c t a t t a a c t g g c g a a c t a c t t a c t t g c t a c g g a c a t c g t t a c c g t t g t t g c a a c g c g t t t g a t a a t t g a c c g c t t g a t g a a t g a

6660 200 205 210 215 LASRQQLIDWMEADKVAGPL AmpR LASRQQLIDWMEADKVAGPL c t a g c t t c c c g g c a a c a a t t a a t a g a c t g g a t g g a g g c g g a t a a a g t t g c a g g a c c a c t t g a t c g a a g g g c c g t t g t t a a t t a t c t g a c c t a c c t c c g c c t a t t t c a a c g t c c t g g t g a a

6720 220 225 230 235 LRSALPAGWFIADKSGAGER AmpR LRSALPAGWFIADKSGAGER c t g c g c t c g g c c c t t c c g g c t g g c t g g t t t a t t g c t g a t a a a t c t g g a g c c g g t g a g c g t g a c g c g a g c c g g g a a g g c c g a c c g a c c a a a t a a c g a c t a t t t a g a c c t c g g c c a c t c g c a

SacII 6780 240 245 250 255 GSRGIIAALGPDGKPSRIVV AmpR GSRGIIAALGPDGKPSRIVV g g a a g c c g c g g t a t c a t t g c a g c a c t g g g g c c a g a t g g t a a g c c c t c c c g t a t c g t a g t t c c t t c g g c g c c a t a g t a a c g t c g t g a c c c c g g t c t a c c a t t c g g g a g g g c a t a g c a t c a a

6840 260 265 270 275 IYTTGSQATMDERNRQIAEI AmpR IYTTGSQATMDERNRQIAEI a t c t a c a c g a c g g g g a g t c a g g c a a c t a t g g a t g a a c g a a a t a g a c a g a t c g c t g a g a t a t a g a t g t g c t g c c c c t c a g t c c g t t g a t a c c t a c t t g c t t t a t c t g t c t a g c g a c t c t a t

6900 280 285 GASLIKHW* AmpR GASLIKHW* g g t g c c t c a c t g a t t a a g c a t t g g t a a c t g t c a g a c c a a g t t t a c t c a t a t a t a c t t t a g c c a c g g a g t g a c t a a t t c g t a a c c a t t g a c a g t c t g g t t c a a a t g a g t a t a t a t g a a a t c

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

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

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

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

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

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

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

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

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

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

7560 ori G G G A A A C G C C T G G T A T C T T T pBR322ori-F g g a g a g c g c a c g a g g g a g c t t c c a g g g g g a a a c g c c t g g t a t c t t t a t a g t c c t g t c g g g c c t c t c g c g t g c t c c c t c g a a g g t c c c c c t t t g c g g a c c a t a g a a a t a t c a g g a c a g c c c

7620 ori t t t c g c c a c c t c t g a c t t g a g c g t c g a t t t t t g t g a t g c t c g t c a g g g g g g c g g a g c c t a a a a g c g g t g g a g a c t g a a c t c g c a g c t a a a a a c a c t a c g a g c a g t c c c c c c g c c t c g g a t

7680 ori t g g a a a a a c g c c a g c a a c g c g g c c t t t t t a c g g t t c c t g g c c t t t t g c t g g c c t t t t g c t a c c t t t t t g c g g t c g t t g c g c c g g a a a a a t g c c a a g g a c c g g a a a a c g a c c g g a a a a c g a

Acc65IKpnI 7740 CMV enhancer c a c a t g c t a g a g g t a c c c g t t a c a t a a c t t a c g g t a a a t g g c c c g c c t g g c t g a c c g c c c g t g t a c g a t c t c c a t g g g c a a t g t a t t g a a t g c c a t t t a c c g g g c g g a c c g a c t g g c g g g

7800 CMV enhancer a a c g a c c c c c g c c c a t t g a c g t c a a t a g t a a c g c c a a t a g g g a c t t t c c a t t g a c g t c a a t t g c t g g g g g c g g g t a a c t g c a g t t a t c a t t g c g g t t a t c c c t g a a a g g t a a c t g c a g t t

NdeI 7860 CMV enhancer t g g g t g g a g t a t t t a c g g t a a a c t g c c c a c t t g g c a g t a c a t c a a g t g t a t c a t a t g c c a a c c c a c c t c a t a a a t g c c a t t t g a c g g g t g a a c c g t c a t g t a g t t c a c a t a g t a t a c g g t

7920 CMV enhancer MARLALCPV 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 g t g c c c a g t a c 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 c a c g g g t c a t g

SnaBI 7980 CMV enhancer HDLMGLSYLAVHLRISHRYY 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 c t a t t a c c 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 g a t a a t g g

8040 CMV enhancer HGRGEPHVLLHSPHLPPLPT a t g g t c g a g g t g a g c c c c a c g t t c t g c t t c a c t c t c c c c a t c t c c c c c c c c t c c c c a c c c t a c c a g c t c c a c t c g g g g t g c a a g a c g a a g t g a g a g g g g t a g a g g g g g g g g a g g g g t g g g

8100 PNFVFIYFLIILCSDGGGGG c c a a t t t t g t a t t t a t t t a t t t t t t a a t t a t t t t g t g c a g c g a t g g g g g c g g g g g g g g g g g g t t a a a a c a t a a a t a a a t a a a a a a t t a a t a a a a c a c g t c g c t a c c c c c g c c c c c c c c c c

8160 GGXRARXGGGGGGGGGGGGG g g g g g g b g c g c g c c a g g s g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g g c c c c c c v c g c g c g g t c c s c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c c

3ʹ 5ʹ 8191 GGGGGGGGGGX g g g g g g g g g g g g g g g g g g g g g g g g g g g g g s g c c c c c c c c c c c c c c c c c c c c c c c c c c c c c s c

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.