Addgene PET-B-Sniper SpCas9-NLS-6xHis Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: PET-B-Sniper SpCas9-NLS-6xHis Sequencing Result


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BspDI * - ClaI * (9199) SgrAI (9155) pBRrevBam (9113 .. 9132) LacI-R (8791 .. 8810) BstEII (8292) BssHII (8063) HincII - HpaI (7972) pGEX 3' (6968 .. 6990) PflFI - Tth111I (6868) BstZ17I (6843) AccI (6842) pRS-marker (6808 .. 6827) BspQI - SapI (6730) L4440 (6711 .. 6728) PciI (6613) pBR322ori-F (6458 .. 6477) Amp-R (5558 .. 5577) PvuI (5378) FspI (5230) BsaI (5069) SspI (4803) PsiI (4723) F1ori-F (4662 .. 4683) T7 (1 .. 20) T7 promoter XbaI (47) RBS NcoI (86) BglII (338) BmgBI (627) NheI (1699) BmtI (1703) EcoRV (1941) NruI (2377) PmlI (2877) KflI (3444) EagI (3690) BsmI (3707) FseI (4199) 6xHis AvrII (4252) T7 Term (4267 .. 4285) F1ori-R (4452 .. 4471) PET-B-Sniper SpCas9-NLS-6xHis 9219 bp

Sequence View

XbaI 5ʹ 3ʹ 60 T7 promoter lac operator RBS T A A T A C G A C T C A C T A T A G G G T7 t a a t a c g a c t c a c t a t a g g g g a a t t g t g a g c g g a t a a c a a t t c c c c t c t a g a a a t a a t t t a t t a t g c t g a g t g a t a t c c c c t t a a c a c t c g c c t a t t g t t a a g g g g a g a t c t t t a t t a a a

NcoI 120 1 5 10 MDKKYSIGLDI RBS Cas9 MDKKYSIGLDI *SPSIGHVLLVADAQVD t g t t t a a c t t t a a g a a g g a g a t a t a c c a t g g a c a a g a a g t a c a g c a t c g g c c t g g a c a t c a c a a a t t g a a a t t c t t c c t c t a t a t g g t a c c t g t t c t t c a t g t c g t a g c c g g a c c t g t a g

180 15 20 25 30 GTNSVGWAVITDEYKVPSKK Cas9 GTNSVGWAVITDEYKVPSKK AGVRHAPGHDGVLVLHGALF 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 a 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 t t

240 35 40 45 50 FKVLGNTDRHSIKKNLIGAL Cas9 FKVLGNTDRHSIKKNLIGAL ELHQAVGVPVADLLVQDSGQ 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 g 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 a c

300 55 60 65 70 LFDSGETAEATRLKRTARRR Cas9 LFDSGETAEATRLKRTARRR QEVAAFCGLGGPQLSGGSSS 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 g a 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 c t

BglII 360 75 80 85 90 YTRRKNRICYLQEIFSNEMA Cas9 YTRRKNRICYLQEIFSNEMA VGSPLVPDAIQLLDEAVLHG 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 c 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 g g

420 95 100 105 110 KVDDSFFHRLEESFLVEEDK Cas9 KVDDSFFHRLEESFLVEEDK LHVVAEEVSQFLGEQHFLIL 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 a 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 t c

480 115 120 125 130 KHERHPIFGNIVDEVAYHEK Cas9 KHERHPIFGNIVDEVAYHEK LVLPVGDEAVDHVLHGVVLL 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 a g 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 t c

540 135 140 145 150 YPTIYHLRKKLVDSTDKADL Cas9 YPTIYHLRKKLVDSTDKADL VGGDVVQSLFQHVAGVLGVQ 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 g 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 a c

600 155 160 165 170 RLIYLALAHMIKFRGHFLIE Cas9 RLIYLALAHMIKFRGHFLIE PQDIQGQGVHDLEPAVEQDL 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 g 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 t c

BmgBI 660 175 180 185 190 GDLNPDNSDVDKLFIQLVQT Cas9 GDLNPDNSDVDKLFIQLVQT AVQVGVVAVHVLQEDLQHLG 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 c 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 g g

720 195 200 205 210 YNQLFEENPINASGVDAKAI Cas9 YNQLFEENPINASGVDAKAI VVLQELFVGDVGAAHVGLGD 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 t c 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 a g

780 215 220 225 230 LSARLSKSRRLENLIAQLPG Cas9 LSARLSKSRRLENLIAQLPG QRGSQALASPQFIQDGLQGA 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 g c 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 c g

840 235 240 245 250 EKKNGLFGNLIALSLGLTPN Cas9 EKKNGLFGNLIALSLGLTPN LLLIAQESVQNGQAQAQGGV 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 a c 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 t g

900 255 260 265 270 FKSNFDLAEDAKLQLSKDTY Cas9 FKSNFDLAEDAKLQLSKDTY ELAVEVQGLIGFQLQALVGV 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 a c 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 t g

960 275 280 285 290 DDDLDNLLAQIGDQYADLFL Cas9 DDDLDNLLAQIGDQYADLFL VVVQVVQQGLDAVLVGVQKQ 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 a c

1020 295 300 305 310 AAKNLSDAILLSDILRVNTE Cas9 AAKNLSDAILLSDILRVNTE GGLVQGVGDQQAVDQSHVGL 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 g 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 t c

1080 315 320 325 330 ITKAPLSASMIKRYDEHHQD Cas9 ITKAPLSASMIKRYDEHHQD DGLGGQAGRHDLSVVLVVLV 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 a 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 t g

1140 335 340 345 350 LTLLKALVRQQLPEKYKEIF Cas9 LTLLKALVRQQLPEKYKEIF QGQQFSEHPLLQRLLVFLNE 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 c 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 a g

1200 355 360 365 370 FDQSKNGYAGYIDGGASQEE Cas9 FDQSKNGYAGYIDGGASQEE EVLALVAVGAVNVASGALFL 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 t c

1260 375 380 385 390 FYKFIKPILEKMDGTEELLV Cas9 FYKFIKPILEKMDGTEELLV EVLEDLGDQFLHVAGLFQEH 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 g 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 a c

1320 395 400 405 410 KLNREDLLRKQRTFDNGSIP Cas9 KLNREDLLRKQRTFDNGSIP LQVSLVQQPLLPGEVVAADG 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 c 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 g g

1380 415 420 425 430 HQIHLGELHAILRRQEDFYP Cas9 HQIHLGELHAILRRQEDFYP VLDVQSLQVGNQPPLFIKVW 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 c a 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 g t

1440 435 440 445 450 FLKDNREKIEKILTFRIPYY Cas9 FLKDNREKIEKILTFRIPYY EQLVVPFLDLLDQGEADGVV 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 a c 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 t g

1500 455 460 465 470 VGPLARGNSRFAWMTRKSEE Cas9 VGPLARGNSRFAWMTRKSEE HARQGPSVASEGPHGSLALF 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 a a 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 t t

1560 475 480 485 490 TITPWNFEEVVDKGASAQSF Cas9 TITPWNFEEVVDKGASAQSF GDGGPVELFHHVLASGGLAE 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 c 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 a g

1620 495 500 505 510 IERMTNFDKNLPNEKVLPKH Cas9 IERMTNFDKNLPNEKVLPKH DLPHGVEILVQGVLLHQGLV 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 c 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 t g

1680 515 520 525 530 SLLYEYFTVYNELTKVKYVT Cas9 SLLYEYFTVYNELTKVKYVT AQQVLVEGHIVLQGFHFVHG 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 c c 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 g g

BmtINheI 1740 535 540 545 550 EGMRKPASLSGEQKKAIVDL Cas9 EGMRKPASLSGEQKKAIVDL LSHSLGSAQAALLFLGDHVQ g a g g g a a t g a g a a a g c c c g c t a g c c t g a g c g g c g a g c a g a a a a a g g c c a t c g t g g a c c t g c t c c c t t a c t c t t t c g g g c g a t c g g a c t c g c c g c t c g t c t t t t t c c g g t a g c a c c t g g a c

1800 555 560 565 570 LFKTNRKVTVKQLKEDYFKK Cas9 LFKTNRKVTVKQLKEDYFKK QELGVPFHGHLLQFLVVELF 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 a 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 t t

1860 575 580 585 590 IECFDSVEISGVEDRFNASL Cas9 IECFDSVEISGVEDRFNASL DLAEVGHFDGAHFIPEVGGQ 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 g 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 a c

1920 595 600 605 610 GTYHDLLKIIKDKDFLDNEE Cas9 GTYHDLLKIIKDKDFLDNEE ACVVIQQFNDLVLVEQVILF 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 a 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 t t

EcoRV 1980 615 620 625 630 NEDILEDIVLTLTLFEDREM Cas9 NEDILEDIVLTLTLFEDREM VLVNQFIDHQGQCQKLVSLH 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 g 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 a c

2040 635 640 645 650 IEERLKTYAHLFDDKVMKQL Cas9 IEERLKTYAHLFDDKVMKQL *AWRNSSLTIFCS DLFPQFGIGVQEVVFHHLLQ 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 t g 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 a c

2100 655 660 665 670 KRRRYTGWGRLSRKLINGIR Cas9 KRRRYTGWGRLSRKLINGIR FRRLYVPQPLSLRFSILPMR LPPSVGAPAPQAPLQDVADP 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 g g c a t c c g g 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 c c g t a g g c c

2160 675 680 685 690 DKQSGKTILDFLKSDGFANR Cas9 DKQSGKTILDFLKSDGFANR SLCDPLVIRSKRFDSPKALL VLLGALCDQIEQLGVAEGVS 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 g a 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 c t

2220 695 700 705 710 NFMQLIHDDSLTFKEDIQKA Cas9 NFMQLIHDDSLTFKEDIQKA FKMCSIWSSLRVKLSSMWFA VEHLQDVVVAQGKFLVDLFG 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 a g a a a g c c 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 g t c t t t c g g

2280 715 720 725 730 QVSGQGDSLHEHIANLAGSP Cas9 QVSGQGDSLHEHIANLAGSP WTDPWPSLRCSCMALRAPLG LHGALAIAQVLVNGIQGAAG 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 c c 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 g g

2340 735 740 745 750 AIKKGILQTVKVVDELVKVM Cas9 AIKKGILQTVKVVDELVKVM AMLFPMRCVTFTTSSSTFTI GNLLADQLCHLHHVLEHFHH 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 t 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 a c

NruI 2400 755 760 765 770 GRHKPENIVIEIARENQTTQ Cas9 GRHKPENIVIEIARENQTTQ PRCLGSFMTISIALSFWVVW APVLGLVDHDFDRSLVLGGL 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 c g c g a g a g a g a a c c a g a c c a c c c a g 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 g c g c t c t c t c t t g g t c t g g t g g g t c

2460 775 780 785 790 KGQKNSRERMKRIEEGIKEL Cas9 KGQKNSRERMKRIEEGIKEL FPCFFLRSLIFRISSPM LSLLVAALSHLPDFLADFLQ 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 g 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 a c

2520 795 800 805 810 GSQILKEHPVENTQLQNEKL Cas9 GSQILKEHPVENTQLQNEKL AALDQFFVGHFVGLQLVLLQ 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 t g 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 a c

2580 815 820 825 830 YLYYLQNGRDMYVDQELDIN Cas9 YLYYLQNGRDMYVDQELDIN VQVVQLIPPIHVHVLFQVDV 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 t g

2640 835 840 845 850 RLSDYDVDHIVPQSFLKDDS Cas9 RLSDYDVDHIVPQSFLKDDS PQGVVIHVMDHRLAKQLVVG 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 c c 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 g g

2700 855 860 865 870 IDNKVLTRSDKNRGKSDNVP Cas9 IDNKVLTRSDKNRGKSDNVP DVVLHQGSAVLVPALAVVHG 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 c c 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 g g

2760 875 880 885 890 SEEVVKKMKNYWRQLLNANL Cas9 SEEVVKKMKNYWRQLLNANL GFLDHLLHLVVPPLQQVRIQ 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 g a a t t t 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 c t t a a a c

2820 895 900 905 910 ITQRKFDNLTKAERGGLSEL Cas9 ITQRKFDNLTKAERGGLSEL NGLSLEVIQGLGLSAAQAFQ 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 t g 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 a c

PmlI 2880 915 920 925 930 DKAGFIKRQLVETRQITKHV Cas9 DKAGFIKRQLVETRQITKHV ILGAEDLSLQHFGPLDCLVH 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 t g 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 a c

2940 935 940 945 950 AQILDSRMNTKYDENDKLIR Cas9 AQILDSRMNTKYDENDKLIR CLDQVGPHVSLVVLIVLQDP 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 g g 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 c c

3000 955 960 965 970 EVKVITLKSKLVSDFRKDFQ Cas9 EVKVITLKSKLVSDFRKDFQ FHFHDGQLGLQHGIEPLIEL 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 a g 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 t c

3060 975 980 985 990 FYKVREINNYHHAHDAYLNA Cas9 FYKVREINNYHHAHDAYLNA KVFHALDVVVVVGVVRVQVG 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 c 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 g g

3120 995 1000 1005 1010 VVGTALIKKYPKLGGKVYDV Cas9 VVGTALIKKYPKLGGKVYDV DHSGGQDFLVRLQTALHVIY 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 t a 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 a t

3180 1015 1020 1025 1030 RKMIAKSEQEIGKATAKYFF Cas9 RKMIAKSEQEIGKATAKYFF PLHDGLALLFDALSGGLVEE c g g a a g a t g a t c g c c a a g a g c g a g c a g g a a a t c g g c a a g g c t a c c g c c a a g t a c t t c t t c g c c t t c t a c t a g c g g t t c t c g c t c g t c c t t t a g c c g t t c c g a t g g c g g t t c a t g a a g a a g

3240 1035 1040 1045 1050 YSNIMNFFKTEITLANGEIR Cas9 YSNIMNFFKTEITLANGEIR VAVDHVKELGLNGQGVALDP t a c a g c a a c a t c a t g a a c t t t t t c a a g a c c g a g a t t a c c c t g g c c a a c g g c g a g a t c c g g a t g t c g t t g t a g t a c t t g a a a a a g t t c t g g c t c t a a t g g g a c c g g t t g c c g c t c t a g g c c

3300 1055 1060 1065 1070 KRPLIETNGETGEIVWDKGR Cas9 KRPLIETNGETGEIVWDKGR LPRQDLCVAFGPLDHPILAP a a g c g g c c t c t g a t c g a g a c a a a c g g c g a a a c c g g g g a g a t c g t g t g g g a t a a g g g c c g g t t c g c c g g a g a c t a g c t c t g t t t g c c g c t t t g g c c c c t c t a g c a c a c c c t a t t c c c g g c c

3360 1075 1080 1085 1090 DFATVRKVLSMPQVNIVKKT Cas9 DFATVRKVLSMPQVNIVKKT IKGGHPFHQAHGLHIDHFLG g a t t t t g c c a c c g t g c g g a a a g t g c t g a g c a t g c c c c a a g t g a a t a t c g t g a a a a a g a c c c t a a a a c g g t g g c a c g c c t t t c a c g a c t c g t a c g g g g t t c a c t t a t a g c a c t t t t t c t g g

3420 1095 1100 1105 1110 EVQTGGFSKESILPKRNSDK Cas9 EVQTGGFSKESILPKRNSDK LHLCAAEAFLRDQGLPVAIL g a g g t g c a g a c a g g c g g c t t c a g c a a a g a g t c t a t c c t g c c c a a g a g g a a c a g c g a t a a g c t c c a c g t c t g t c c g c c g a a g t c g t t t c t c a g a t a g g a c g g g t t c t c c t t g t c g c t a t t c

KflI 3480 1115 1120 1125 1130 LIARKKDWDPKKYGGFDSPT Cas9 LIARKKDWDPKKYGGFDSPT QDGSLLVPVRLLVAAEVAGG c t g a t c g c c a g a a a g a a g g a c t g g g a c c c t a a g a a g t a c g g c g g c t t c g a c a g c c c c a c c g a c t a g c g g t c t t t c t t c c t g a c c c t g g g a t t c t t c a t g c c g c c g a a g c t g t c g g g g t g g

3540 1135 1140 1145 1150 VAYSVLVVAKVEKGKSKKLK Cas9 VAYSVLVVAKVEKGKSKKLK HGIRHQHHGFHFLALGLFQL g t g g c c t a t t c t g t g c t g g t g g t g g c c a a a g t g g a a a a g g g c a a g t c c a a g a a a c t g a a g c a c c g g a t a a g a c a c g a c c a c c a c c g g t t t c a c c t t t t c c c g t t c a g g t t c t t t g a c t t c

3600 1155 1160 1165 1170 SVKELLGITIMERSSFEKNP Cas9 SVKELLGITIMERSSFEKNP THFLQQPDGDHFSAAELLIG a g t g t g a a a g a g c t g c t g g g g a t c a c c a t c a t g g a a a g a a g c a g c t t c g a g a a g a a t c c c t c a c a c t t t c t c g a c g a c c c c t a g t g g t a g t a c c t t t c t t c g t c g a a g c t c t t c t t a g g g

3660 1175 1180 1185 1190 IDFLEAKGYKEVKKDLIIKL Cas9 IDFLEAKGYKEVKKDLIIKL DVKQFGLAVFFHFLVQDDLQ a t c g a c t t t c t g g a a g c c a a g g g c t a c a a a g a a g t g a a a a a g g a c c t g a t c a t c a a g c t g t a g c t g a a a g a c c t t c g g t t c c c g a t g t t t c t t c a c t t t t t c c t g g a c t a g t a g t t c g a c

BsmIEagI 3720 1195 1200 1205 1210 PKYSLFELENGRKRMLASAG Cas9 PKYSLFELENGRKRMLASAG RLVGQELQFVAPLSHQGRGA c c t a a g t a c t c c c t g t t c g a g c t g g a a a a c g g c c g g a a g a g a a t g c t g g c c t c t g c c g g c g g a t t c a t g a g g g a c a a g c t c g a c c t t t t g c c g g c c t t c t c t t a c g a c c g g a g a c g g c c g

3780 1215 1220 1225 1230 ELQKGNELALPSKYVNFLYL Cas9 ELQKGNELALPSKYVNFLYL FQLLSVFQGQGGFIHVEQVQ g a a c t g c a g a a g g g a a a c g a a c t g g c c c t g c c c t c c a a a t a t g t g a a c t t c c t g t a c c t g c t t g a c g t c t t c c c t t t g c t t g a c c g g g a c g g g a g g t t t a t a c a c t t g a a g g a c a t g g a c

3840 1235 1240 1245 1250 ASHYEKLKGSPEDNEQKQLF Cas9 ASHYEKLKGSPEDNEQKQLF GAVILLQLAGGLIILLFLQK g c c a g c c a c t a t g a g a a g c t g a a g g g c t c c c c c g a g g a t a a t g a g c a g a a a c a g c t g t t t c g g t c g g t g a t a c t c t t c g a c t t c c c g a g g g g g c t c c t a t t a c t c g t c t t t g t c g a c a a a

3900 1255 1260 1265 1270 VEQHKHYLDEIIEQISEFSK Cas9 VEQHKHYLDEIIEQISEFSK HFLVLVVQVLDDLLDALEGL g t g g a a c a g c a c a a g c a c t a c c t g g a c g a g a t c a t c g a g c a g a t c a g c g a g t t c t c c a a g c a c c t t g t c g t g t t c g t g a t g g a c c t g c t c t a g t a g c t c g t c t a g t c g c t c a a g a g g t t c

3960 1275 1280 1285 1290 RVILADANLDKVLSAYNKHR Cas9 RVILADANLDKVLSAYNKHR SHDQGVSIQVFHQGGVVLVP a g a g t g a t c c t g g c c g a c g c t a a t c t g g a c a a a g t g c t g t c c g c c t a c a a c a a g c a c c g g t c t c a c t a g g a c c g g c t g c g a t t a g a c c t g t t t c a c g a c a g g c g g a t g t t g t t c g t g g c c

4020 1295 1300 1305 1310 DKPIREQAENIIHLFTLTNL Cas9 DKPIREQAENIIHLFTLTNL ILGDSLLGLIDDVQKGQGIQ g a t a a g c c c a t c a g a g a g c a g g c c g a g a a t a t c a t c c a c c t g t t t a c c c t g a c c a a t c t g c t a t t c g g g t a g t c t c t c g t c c g g c t c t t a t a g t a g g t g g a c a a a t g g g a c t g g t t a g a c

4080 1315 1320 1325 1330 GAPAAFKYFDTTIDRKRYTS Cas9 GAPAAFKYFDTTIDRKRYTS SGRGGELVKVGGDVPLPVGA g g a g c c c c t g c c g c c t t c a a g t a c t t t g a c a c c a c c a t c g a c c g g a a g a g g t a c a c c a g c c c t c g g g g a c g g c g g a a g t t c a t g a a a c t g t g g t g g t a g c t g g c c t t c t c c a t g t g g t c g

4140 1335 1340 1345 1350 TKEVLDATLIHQSITGLYET Cas9 TKEVLDATLIHQSITGLYET GFLHQVGGQDVLADGAQVLC a c c a a a g a g g t g c t g g a c g c c a c c c t g a t c c a c c a g a g c a t c a c c g g c c t g t a c g a g a c a t g g t t t c t c c a c g a c c t g c g g t g g g a c t a g g t g g t c t c g t a g t g g c c g g a c a t g c t c t g t

FseI 4200 1355 1360 RIDLSQLGGD 1 5 10 KRPAATKKAG Cas9 nucleoplasmin NLS RIDLSQLGGDKRPAATKKAG PDVQRLQSAVFPRRGRFLGA c g g a t c g a c c t g t c t c a g c t g g g a g g c g a c a a a a g g c c g g c g g c c a c g a a a a a g g c c g g c g c c t a g c t g g a c a g a g t c g a c c c t c c g c t g t t t t c c g g c c g c c g g t g c t t t t t c c g g c c g

AvrII 4260 15 QAKKKK MDK 1 5 HHHHHH * nucleoplasmin NLS 6xHis QAKKKKMDKHHHHHH* LCFLFLHILMVVVMVLLRPQ c a g g c a a a a a a g a a a a a g a t g g a t a a g c a t c a c c a c c a c c a t c a c t a a t a a c c t a g g c t g g t c c g t t t t t t c t t t t t c t a c c t a t t c g t a g t g g t g g t g g t a g t g a t t a t t g g a t c c g a c

4320 T7 terminator G G C G A C T C G T T A T T G A T C G T7 Term QWRQAIVLM c t g c c a c c g c t g a g c a a t a a c t a g c a t a a c c c c t t g g g g c c t c t a a a c g g g t c t t g a g g g g a c g g t g g c g a c t c g t t a t t g a t c g t a t t g g g g a a c c c c g g a g a t t t g c c c a g a a c t c c c

4380 T7 terminator f1 ori g t t t t t t g c t g a a a g g a g g a a c t a t a t c c g g a t t g g c g a a t g g g a c g c g c c c t g t a g c g g c a a a a a a c g a c t t t c c t c c t t g a t a t a g g c c t a a c c g c t t a c c c t g c g c g g g a c a t c g c c

4440 f1 ori c g c a t t a a g c g c g g c g g g t g t g g t g g t t a c g c g c a g c g t g a c c g c t a c a c t t g c c a g c g c g c g t a a t t c g c g c c g c c c a c a c c a c c a a t g c g c g t c g c a c t g g c g a t g t g a a c g g t c g c g

4500 f1 ori G A G G A A A G C G A A A G A A G G G A F1ori-R c c t a g c g c c c g c t c c t t t c g c t t t c t t c c c t t c c t t t c t c g c c a c g t t c g c c g g c t t t c c g g a t c g c g g g c g a g g a a a g c g a a a g a a g g g a a g g a a a g a g c g g t g c a a g c g g c c g a a a g g

4560 f1 ori c c g t c a a g c t c t a a a t c g g g g g c t c c c t t t a g g g t t c c g a t t t a g t g c t t t a c g g c a c c t g g c a g t t c g a g a t t t a g c c c c c g a g g g a a a t c c c a a g g c t a a a t c a c g a a a t g c c g t g g a

4620 f1 ori c g a c c c c a a a a a a c t t g a t t a g g g t g a t g g t t c a c g t a g t g g g c c a t c g c c c t g a t a g a c g c t g g g g t t t t t t g a a c t a a t c c c a c t a c c a a g t g c a t c a c c c g g t a g c g g g a c t a t c t g

4680 f1 ori G T G G A C T C T T G T T C C A A A C F1ori-F g g t t t t t c g c c c t t t g a c g t t g g a g t c c a c g t t c t t t a a t a g t g g a c t c t t g t t c c a a a c c c a a a a a g c g g g a a a c t g c a a c c t c a g g t g c a a g a a a t t a t c a c c t g a g a a c a a g g t t t g

PsiI 4740 f1 ori T G G F1ori-F t g g a a c a a c a c t c a a c c c t a t c t c g g t c t a t t c t t t t g a t t t a t a a g g g a t t t t g c c g a t a c c t t g t t g t g a g t t g g g a t a g a g c c a g a t a a g a a a a c t a a a t a t t c c c t a a a a c g g c t a

4800 f1 ori t t c g g c c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a a a a t t t a a c g c g a a t t t t a a c a a a a g c c g g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t t t t t a a a t t g c g c t t a a a a t t g t t

SspI 4860 f1 ori a a t a t t a a c g t t t a c a a t t t c t g g c g g c a c g a t g g 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 t t a t a a t t g c a a a t g t t a a a g a c c g c c g t g c t a c c 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

4920 a c c t a g a t c c t t t t a a a t t a a a a a t g a a g t t t t a a a t c a a t c t a a a g t a t a t a t g a g t a a t g g a t c t a g g a a a a t t t a a t t t t t a c t t c a a a a t t t a g t t a g a t t t c a t a t a t a c t c a t t

4980 285 280 275 *WHKILSAGIEAIQR AmpR *WHKILSAGIEAIQR a c t t g g t c t g a c a g t t a c c a a t g c t t a a t c a g t g a g g c a c c t a t c t c a g c g a t c t g t c t a t g a a c c a g a c t g t c a a t g g t t a c g a a t t a g t c a c t c c g t g g a t a g a g t c g c t a g a c a g a t

5040 270 265 260 255 NREDMTAQSGTTYIVVIRSP AmpR NREDMTAQSGTTYIVVIRSP t t t c g t t c a t c c a t a g t t g c c t g a c t c c c c g t c g t g t a g a t a a c t a c g a t a c g g g a g g g c a a a g c a a g t a g g t a t c a a c g g a c t g a g g g g c a g c a c a t c t a t t g a t g c t a t g c c c t c c c g

BsaI 5100 250 245 240 235 KGDPGLAAIIGRSGREGAGS AmpR KGDPGLAAIIGRSGREGAGS t t a c c a t c t g g c c c c a g t g c t g c a a t g a t a c c g c g a g a c c c a c g c t c a c c g g c t c c a g a t a a t g g t a g a c c g g g g t c a c g a c g t t a c t a t g g c g c t c t g g g t g c g a g t g g c c g a g g t c t a

5160 230 225 220 215 KDAIFWGAPLASRLLPGAVK AmpR KDAIFWGAPLASRLLPGAVK t t a t c a g c a a t a a a c c a g c c a g c c g g a a g g g c c g a g c g c a g a a g t g g t c c t g c a a c t t t a a a t a g t c g t t a t t t g g t c g g t c g g c c t t c c c g g c t c g c g t c t t c a c c a g g a c g t t g a a a t

5220 210 205 200 195 DAEMWDILQQRSALTLLEGT AmpR DAEMWDILQQRSALTLLEGT t c c g c c t c c a t c c a g t c t a t t a a t t g t t g c c g g g a a g c t a g a g t a a g t a g t t c g c c a g t t a g g c g g a g g t a g g t c a g a t a a t t a a c a a c g g c c c t t c g a t c t c a t t c a t c a a g c g g t c a a

FspI 5280 190 185 180 175 LLKRLTTAMAVPMTTDREDN AmpR LLKRLTTAMAVPMTTDREDN a a t a g t t t g c g c a a c g t t g t t g c c a t t g c t a c a g g c a t c g t g g t g t c a c g c t c g t c g t t t t t a t c a a a c g c g t t g c a a c a a c g g t a a c g a t g t c c g t a g c a c c a c a g t g c g a g c a g c a a a

5340 170 165 160 155 PIAENLEPEWRDLRTVHDGM AmpR PIAENLEPEWRDLRTVHDGM g g t a t g g c t t c a t t c a g c t c c g g t t c c c a a c g a t c a a g g c g a g t t a c a t g a t c c c c c a t g c c a t a c c g a a g t a a g t c g a g g c c a a g g g t t g c t a g t t c c g c t c a a t g t a c t a g g g g g t a c

PvuI 5400 150 145 140 135 NHLFATLEKPGGITTLLLNA AmpR NHLFATLEKPGGITTLLLNA t t g t g c a a a a a a g c g g t t a g c t c c t t c g g t c c t c c g a t c g t t g t c a g a a g t a a g t t g g c c a a c a c g t t t t t t c g c c a a t c g a g g a a g c c a g g a g g c t a g c a a c a g t c t t c a t t c a a c c g g

5460 130 125 120 115 ATNDSMTIAASCLERVTMGD AmpR ATNDSMTIAASCLERVTMGD g c a g t g t t a t c a c t c a t g g t t a t g g c a g c a c t g c a t a a t t c t c t t a c t g t c a t g c c a t c c c g t c a c a a t a g t g a g t a c c a a t a c c g t c g t g a c g t a t t a a g a g a a t g a c a g t a c g g t a g g

5520 110 105 100 95 TLHKETVPSYEVLDNQSYHI AmpR TLHKETVPSYEVLDNQSYHI g t a a g a t g c t t t t c t g t g a c t g g t g a g t a c t c a a c c a a g t c a t t c t g a g a a t a g t g t a t g c a t t c t a c g a a a a g a c a c t g a c c a c t c a t g a g t t g g t t c a g t a a g a c t c t t a t c a c a t a c

5580 90 85 80 75 RRGLQEQGADIRSLVAGCLL AmpR A T A A T A C C G C G C C A C A T A G C Amp-R RRGLQEQGADIRSLVAGCLL c g g c g a c c g a g t t g c t c t t g c c c g g c g t c a a t a c g g g a t a a t a c c g c g c c a c a t a g c a g a g c c g c t g g c t c a a c g a g a a c g g g c c g c a g t t a t g c c c t a t t a t g g c g c g g t g t a t c g t c t

5640 70 65 60 55 VKFTSMMPFREEPRFSELIK AmpR VKFTSMMPFREEPRFSELIK a c t t t a a a a g t g c t c a t c a t t g g a a a a c g t t c t t c g g g g c g a a a a c t c t c a a g g a t c t t a t g a a a t t t t c a c g a g t a g t a a c c t t t t g c a a g a a g c c c c g c t t t t g a g a g t t c c t a g a a t

5700 50 45 40 35 GSNLDLEIYGVRAGLQDEAD AmpR GSNLDLEIYGVRAGLQDEAD c c g c t g t t g a g a t c c a g t t c g a t g t a a c c c a c t c g t g c a c c c a a c t g a t c t t c a g c a t c t g g c g a c a a c t c t a g g t c a a g c t a c a t t g g g t g a g c a c g t g g g t t g a c t a g a a g t c g t a g a

5760 30 25 KVKVLTEPH 20 15 signal sequence AFVPLCFAAFF AmpR KVKVLTEPHAFVPLCFAAFF t t t a c t t t c a c c a g c g t t t c t g g g t g a g c a a a a a c a g g a a g g c a a a a t g c c g c a a a a a a g a a a t g a a a g t g g t c g c a a a g a c c c a c t c g t t t t t g t c c t t c c g t t t t a c g g c g t t t t t t c

5820 10 5 1 signal sequence PILAVRFHQISM AmpR AmpR promoter PILAVRFHQISM g g a a t a a g g g c g a c a c g g a a a t g t t g a a t a c t c a t a c t c t t c c t t t t t c a a t c a t g a t t g c c t t a t t c c c g c t g t g c c t t t a c a a c t t a t g a g t a t g a g a a g g a a a a a g t t a g t a c t a a c

5880 AmpR promoter a a g c a t t t a t c a g g g t t a t t g t c t c a t g a g c g g a t a c a t a t t t g a a t g t a t t t a g a a a a a t t c g t a a a t a g t c c c a a t a a c a g a g t a c t c g c c t a t g t a t a a a c t t a c a t a a a t c t t t t t

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

6000 ori a c c c c g t a g a a a a g a t c a a a g g a t c t t c t t g a g a t c c t t t t t t t c t g c g c g t a a t c t g c t t g g g g c a t c t t t t c t a g t t t c c t a g a a g a a c t c t a g g a a a a a a a g a c g c g c a t t a g a c g a

6060 ori g c t t g c a a a c a a a a a a a c c a c c g c t a c c a g c g g t g g t t t g t t t g c c g g a t c a a g a g c t a c c g a a c g t t t g 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

6120 ori c a a c t c t t t t t c c g a a g g t a a c t g g c t t c a g c a g a g c g c a g a t a c c a a a t a c t g t c c t t c g t t g a g a a a a a g g c t t c c a t t g a c c g a a g t c g t c t c g c g t c t a t g g t t t a t g a c a g g a a g

6180 ori t a g t g t a g c c g t a g t t a g g c c a c c a c t t c a a g a a c t c t g t a g c a c c g c c t a c a t a c c t c g a t c a c a t c g g 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

6240 ori c t c t g c t a a t c c t g t t a c c a g t g g c t g c t g c c a g t g g c g a t a a g t c g t g t c t t a c c g g g t g a g a c g a t t a g g a c a a t g g t c a c c g a c g a c g g t c a c c g c t a t t c a g c a c a g a a t g g c c c a

6300 ori t g g a c t c a a g a c g a t a g t t a c c g g a t a a g g c g c a g c g g t c g g g c t g a a c g g g g g g t t c g t a c c t g a g t t c t g c t a t c a a t g g c c t a t t c c g c g t c g c c a g c c c g a c t t g c c c c c c a a g c a

6360 ori g c a c a c a g c c c a g c t t g g a g c g a a c g a c c t a c a c c g a a c t g a g a t a c c t a c a g c g t g a g c c g t g t g t c g g g t c g a a c c t c g c t t g c t g g a t g t g g c t t g a c t c t a t g g a t g t c g c a c t c g

6420 ori t a t g a g a a a g c g c c a c g c t t c c c g a a g g g a g a a a g g c g g a c a g g t a t c c g g t a a g c g g c a a t a c t c t t t c g c g g t g c g a a g g g c t t c c c t c t t t c c g c c t g t c c a t a g g c c a t t c g c c g t

6480 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 g t c g g a a c a g g a g a g c g c a c g a g g g a g c t c 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 c c c a g c c t t g t c c t c t c g c g t g c t c c c t c g a g 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

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

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

PciI 6660 g g c c t t t t g c t c a c a t g t t c t t t c c t g c g t t a t c c c c t g a t t c t g t g g a t a a c c g t a t t a c c g g a a a a c g a g t g t a c a a g a a a g g a c g c a a t a g g g g a c t a a g a c a c c t a t t g g c a t a a t

6720 A G C G A G T C A G L4440 c c g c c t t t g a g t g a g c t g a t a c c g c t c g c c g c a g c c g a a c g a c c g a g c g c a g c g a g t c a g g g c g g a a a c t c a c t c g a c t a t g g c g a g c g g c g t c g g c t t g c t g g c t c g c g t c g c t c a g t c

BspQISapI 6780 bom T G A G C G A G L4440 t g a g c g a g g a a g c g g a a g a g c g c c t g a t g c g g t a t t t t c t c c t t a c g c a t c t g t g c g g t a a c t c g c t c c t t c g c c t t c t c g c g g a c t 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

6840 bom A T G T T A G A C G A G A C T A C G G C pRS-marker t t t c a c a c c g c a a t g g t g c a c t c t c a g t a c a a t c t g c t c t g a t g c c g c a t a g t t a a g c c a a a a g t g t g g c g t t a c c a c g t g a g a g t c a t g t t a g a c g a g a c t a c g g c g t a t c a a t t c g g t

Tth111IPflFIAccIBstZ17I 6900 bom g t a t a c a c t c c g c t a t c g c t a c g t g a c t g g g t c a t g g c t g c g c c c c g a c a c c c g c c a a c a c a t a t g t g a g g c g a t a g c g a t g c a c t g a c c c a g t a c c g a c g c g g g g c t g t g g g c g g t t g t

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

7020 60 55 *LNEGDDGFRAL rop C C G G G A G C T G C A T G T G T C A G A G G pGEX 3' a c c g t c t c c g g g a g c t g c a t g t g t c a g a g g t t t t c a c c g t c a t c a c c g a a a c g c g c g a g g t g g c a g a g g c c 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 c

7080 50 45 40 35 CSRYLEDAHDHLSECIDAQE rop c a g c t g c g g t a a a g c t c a t c a g c g t g g t c g t g a a g c g a t t c a c a g a t g t c t g c c t g t t c a g t c g a c g c c a t t t c g a g t a g t c g c a c c a g c a c t t c g c t a a g t g t c t a c a g a c g g a c a a g t

7140 30 25 20 15 DADLENLKELLTLTQSRIFR rop t c c g c g t c c a g c t c g t t g a g t t t c t c c a g a a g c g t t a a t g t c t g g c t t c t g a t a a a g c g g a g g c g c a g g t c g a g c a a c t c a a a g a g g t c t t c g c a a t t a c a g a c c g a a g a c t a t t t c g c c

7200 10 5 1 AMNLATKEQKTM rop g c c a t g t t a a g g g c g g t t t t t t c c t g t t t g g t c a c t g a t g c c t c c g t g t a a g g g g g a t t t c g g t a c a a t t c c c g c c a a a a a a g g a c a a a c c a g t g a c t a c g g a g g c a c a t t c c c c c t a a a

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

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

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

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

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

7560 g t t g c t c a g g t c g c a g a c g t t t t g c a g c a g c a g t c g c t t c a c g t t c g c t c g c g t a t c g g t c a a c g a g t c c a g c g t c t g c a a a a c g t c g t c g t c a g c g a a g t g c a a g c g a g c g c a t a g c c a

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

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

7740 c t c a a g g g c a t c g g t c g a g a t c c c g g t g c c t a a t g a g t g a g c t a a c t t a c a t t a a t t g c g g a g t t c c c g t a g c c a g c t c t a g g g c c a c g g a t t a c t c a c t c g a t t g a a t g t a a t t a a c g c

7800 360 355 350 345 *QGSELRSVQRALQMLSD lacI *QGSELRSVQRALQMLSD t t g c g c t c a c t g c c c g c t t t c c a g t c g g g a a a c c t g t c g t g c c a g c t g c a t t a a t g a a t c a a c g c g a g t g a c g g g c g a a a g g t c a g c c c t t t g g a c a g c a c g g t c g a c g t a a t t a c t t a g

7860 340 335 330 325 ALARPSATQTNPALTTKRKV lacI ALARPSATQTNPALTTKRKV g g c c a a c g c g c g g g g a g a g g c g g t t t g c g t a t t g g g c g c c a g g g t g g t t t t t c t t t t c a c c c g g t t g c g c g c c c c t c t c c g c c a a a c g c a t a a c c c g c g g t c c c a c c a a a a a g a a a a g t g

7920 320 315 310 305 LSVPLLQNGKVAQGQSLQLL lacI LSVPLLQNGKVAQGQSLQLL c a g t g a g a c g g g c a a c a g c t g a t t g c c c t t c a c c g c c t g g c c c t g a g a g a g t t g c a g c a a g t c a c t c t g c c c g t t g t c g a c t a a c g g g a a g t g g c g g a c c g g g a c t c t c t c a a c g t c g t t

HincIIHpaI 7980 300 295 290 285 RDVSTQGLLRFDQKITTLPP lacI RDVSTQGLLRFDQKITTLPP g c g g t c c a c g c t g g t t t g c c c c a g c a g g c g a a a a t c c t g t t t g a t g g t g g t t a a c g g c g g c g c c a g g t g c g a c c a a a c g g g g t c g t c c g c t t t t a g g a c a a a c t a c c a c c a a t t g c c g c c

8040 280 275 270 265 IYCSSDETDDYGVVSIDAGV lacI IYCSSDETDDYGVVSIDAGV g a t a t a a c a t g a g c t g t c t t c g g t a t c g t c g t a t c c c a c t a c c g a g a t g t c c g c a c c a a c c t a t a t t g t a c t c g a c a g a a g c c a t a g c a g c a t a g g g t g a t g g c t c t a c a g g c g t g g t t g

BssHII 8100 260 255 250 245 RLGSETIARMAGLAMQDNAV lacI RLGSETIARMAGLAMQDNAV g c g c a g c c c g g a c t c g g t a a t g g c g c g c a t t g c g c c c a g c g c c a t c t g a t c g t t g g c a a c c g c g t c g g g c c t g a g c c a t t a c c g c g c g t a a c g c g g g t c g c g g t a g a c t a g c a a c c g t t g

8160 240 235 230 225 LMATPVIGENLMQMTQQFGS lacI LMATPVIGENLMQMTQQFGS c a g c a t c g c a g t g g g a a c g a t g c c c t c a t t c a g c a t t t g c a t g g t t t g t t g a a a a c c g g a g t c g t a g c g t c a c c c t t g c t a c g g g a g t a a g t c g t a a a c g t a c c a a a c a a c t t t t g g c c t

8220 220 215 210 205 MASWDGEREAIPQIQNRTLY lacI MASWDGEREAIPQIQNRTLY c a t g g c a c t c c a g t c g c c t t c c c g t t c c g c t a t c g g c t g a a t t t g a t t g c g a g t g a g a t a g t a c c g t g a g g t c a g c g g a a g g g c a a g g c g a t a g c c g a c t t a a a c t a a c g c t c a c t c t a t

8280 200 195 190 185 KHWGALRLRASVSSLPGALL lacI KHWGALRLRASVSSLPGALL t t t a t g c c a g c c a g c c a g a c g c a g a c g c g c c g a g a c a g a a c t t a a t g g g c c c g c t a a c a g a a a t a c g g t c g g t c g g t c t g c g t c t g c g c g g c t c t g t c t t g a a t t a c c c g g g c g a t t g t c

BstEII 8340 180 175 170 165 AIQQHGLAVLHEVGLRTGDE lacI AIQQHGLAVLHEVGLRTGDE c g c g a t t t g c t g g t g a c c c a a t g c g a c c a g a t g c t c c a c g c c c a g t c g c g t a c c g t c t t c g c g c t a a a c g a c c a c t g g g t t a c g c t g g t c t a c g a g g t g c g g g t c a g c g c a t g g c a g a a g

8400 160 155 150 145 HSFIISNIPTQDSVDLFLAP lacI HSFIISNIPTQDSVDLFLAP a t g g g a g a a a a t a a t a c t g t t g a t g g g t g t c t g g t c a g a g a c a t c a a g a a a t a a c g c c g g t a c c c t c t t t t a t t a t g a c a a c t a c c c a c a g a c c a g t c t c t g t a g t t c t t t a t t g c g g c c

8460 140 135 130 125 VNTCAAEVAIADQDDLPYNI lacI VNTCAAEVAIADQDDLPYNI a a c a t t a g t g c a g g c a g c t t c c a c a g c a a t g g c a t c c t g g t c a t c c a g c g g a t a g t t a a t t t g t a a t c a c g t c c g t c g a a g g t g t c g t t a c c g t a g g a c c a g t a g g t c g c c t a t c a a t t a

8520 120 115 110 105 ILGSVRQALLNHVAAKCAEV lacI ILGSVRQALLNHVAAKCAEV g a t c a g c c c a c t g a c g c g t t g c g c g a g a a g a t t g t g c a c c g c c g c t t t a c a g g c t t c g a c c t a g t c g g g t g a c t g c g c a a c g c g c t c t t c t a a c a c g t g g c g g c g a a a t g t c c g a a g c t g

8580 100 95 90 85 GSREVMSVVVSAGLQDARSK lacI * GSREVMSVVVSAGLQDARSK g c c g c t t c g t t c t a c c a t c g a c a c c a c c a c g c t g g c a c c c a g t t g a t c g g c g c g a g a t t t c g g c g a a g c a a g a t g g t a g c t g t g g t g g t g c g a c c g t g g g t c a a c t a g c c g c g c t c t a a a

8640 80 75 70 65 IAAVIQSPAHLALSSTAVGI lacI DGRCNAVARAPGSQLHCRWD IAAVIQSPAHLALSSTAVGI a a t c g c c g c g a c a a t t t g c g a c g g c g c g t g c a g g g c c a g a c t g g a g g t g g c a a c g c c a a t t t a g c g g c g c t g t t a a a c g c t g c c g c g c a c g t c c c g g t c t g a c c t c c a c c g t t g c g g t t a

8700 60 55 50 45 LLSQKGALQQAVRNPIYNLE lacI AVVTQGGTTTGRPQSHLEAG LLSQKGALQQAVRNPIYNLE c a g c a a c g a c t g t t t g c c c g c c a g t t g t t g t g c c a c g c g g t t g g g a a t g t a a t t c a g c t c g t c g t t g c t g a c a a a c g g g c g g t c a a c a a c a c g g t g c g c c a a c c c t t a c a t t a a g t c g a g

8760 40 35 30 25 AMAAEVKERTKASVHSAQNV lacI GDGSGSKGANECFRPQGPEG AM c g c c a t c g c c g c t t c c a c t t t t t c c c g c g t t t t c g c a g a a a c g t g g c t g g c c t g g t t c a c g c g g t a g c g g c g a a g g t g a a a a a g g g c g c a a a a g c g t c t t t g c a c c g a c c g g a c c a a g t g

8820 20 15 10 5 VRSVTQYSVGAYEAVDYLTV lacI G G C A T A C T C T G C G A C A T C G T LacI-R RPFRDSLLCRCVRRCRIVNS c a c g c g g g a a a c g g t c t g a t a a g a g a c a c c g g c a t a c t c t g c g a c a t c g t a t a a c g t t a c g t g c g c c c t t t g c c a g a c t a t t c t c t g t g g c c g t a t g a g a c g c t g t a g c a t a t t g c a a t g

8880 1 PKM lacI lacI promoter TECEGGQISERGPAIMGYRS t g g t t t c a c a t t c a c c a c c c t g a a t t g a c t c t c t t c c g g g c g c t a t c a t g c c a t a c c g c g a c c a a a g t g t a a g t g g t g g g a c t t a a c t g a g a g a a g g c c c g c g a t a g t a c g g t a t g g c g c

8940 lacI promoter LNQAMRHHGPDRREGKHSEQ a a a g g t t t t g c g c c a t t c g a t g g t g t c c g g g a t c t c g a c g c t c t c c c t t a t g c g a c t c c t t t t c c a a a a c g c g g t a a g c t a c c a c a g g c c c t a g a g c t g c g a g a g g g a a t a c g c t g a g g a

9000 MLFCGLLLNLGNLVAAALFP g c a t t a g g a a g c a g c c c a g t a g t a g g t t g a g g c c g t t g a g c a c c g c c g c c g c a a g g a a t g c g t a a t c c t t c g t c g g g t c a t c a t c c a a c t c c g g c a a c t c g t g g c g g c g g c g t t c c t t a c

9060 AHLSIAGLLGGAVPGAVMGV g t g c a t g c a a g g a g a t g g c g c c c a a c a g t c c c c c g g c c a c g g g g c c t g c c a c c a t a c c c a c a c g t a c g t t c c t c t a c c g c g g g t t g t c a g g g g g c c g g t g c c c c g g a c g g t g g t a t g g g t

9120 G G T G A T G T pBRrevBam GFCASM c g c c g a a a c a a g c g c t c a t g a g c c c g a a g t g g c g a g c c c g a t c t t c c c c a t c g g t g a t g t g c g g c t t t g t t c g c g a g t a c t c g g g c t t c a c c g c t c g g g c t a g a a g g g g t a g c c a c t a c a

SgrAI 9180 C G G C G A T A T A G G pBRrevBam c g g c g a t a t a g g c g c c a g c a a c c g c a c c t g t g g c g c c g g t g a t g c c g g c c a c g a t g c g t c g c c g c t a t a t c c g c g g t c g t t g g c g t g g a c a c c g c g g c c a c t a c g g c c g g t g c t a c g c a g

BspDI*ClaI* 3ʹ 5ʹ 9219 c g g c g t a g a g g a t c g a g a t c g a t c t c g a t c c c g c g a a a t g c c g c a t c t c c t a g c t c t a g c t a g a g c t a g g g c g c t t t a

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.