Addgene pspCas9-ABE-3'UTR-sgRNA-ST2-com vector - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pspCas9-ABE-3'UTR-sgRNA-ST2-com vector


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Acc65I (0) AleI (9585) AbsI - PaeR7I - PspXI - XhoI (9493) AgeI (9481) LKO.1 5' (9406 .. 9425) hU6-F (9235 .. 9255) AflIII - PciI (9229) pBR322ori-F (9074 .. 9093) PvuI (7972) Amp-R (7772 .. 7791) pBRforEco (7363 .. 7381) pGEX 3' (7303 .. 7325) pRS-marker (7184 .. 7203) PsiI (7070) F1ori-F (7009 .. 7030) F1ori-R (6799 .. 6818) SbfI (6635) AAV2 ITR BGH-rev (6272 .. 6289) XbaI (6242) EcoRI (6238) BmtI (6106) NheI (6102) FseI (5943) BsaBI * (5309) KflI (5188) KpnI (4) SnaBI (267) NcoI (287) CMV promoter CMV-F (446 .. 466) NotI (530) T7 (537 .. 556) T7 promoter StuI * (822) BspDI * - ClaI * (849) MreI (1487) BglII (2061) PspOMI (3166) ApaI (3170) PasI (3580) EcoRV (3664) PmlI (4600) pspCas9-ABE-3'UTR-sgRNA-ST2-com vector 9613 bp

Sequence View

Acc65IKpnI 5ʹ 3ʹ 60 CMV enhancer 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 a a c g a c c c c c g 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 t t g c t g g g g g c g

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

180 CMV enhancer 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 g t a c g c c c c c t 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 t c a t g c g g g g g a

240 CMV enhancer 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 a t g a c c t t a t g g 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 t a c t g g a a t a c c

NcoISnaBI 300 CMV enhancer CMV promoter 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 a t g g t g a t g c g g 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 t a c c a c t a c g c c

360 CMV promoter t t t t g g c a g t a c a t c a a t g g g c g t g g a t a g c g g t t t g a c t c a c g g g g a t t t c c a a g t c t c a a a a c c g t c a t g t a g t t a c c c g c a c c t a t c g c c a a a c t g a g t g c c c c t a a a g g t t c a g a g

420 CMV promoter c a c c c c a t t g a c g t c a a t g g g a g t t t g t t t t g g c a c c a a a a t c a a c g g g a c t t t c c a a a a g t g g g g t a a c t g c a g t t a c c c t c a a a c a a a a c c g t g g t t t t a g t t g c c c t g a a a g g t t t t

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

NotI 540 CMV promoter T7 promoter T A A T T7 t a t a t a a g c a g a g c t g g t t t a g t g a a c c g t c a g a t c c g c t a g a g a t c c g c g g c c g c t a a t a t a t a t t c g t c t c g a c c a a a t c a c t t g g c a g t c t a g g c g a t c t c t a g g c g c c g g c g a t t a

600 T7 promoter A C G A C T C A C T A T A G G G T7 MKRTADGSEFE a c g a c t c a c t a t a g g g a g a g c c g c c a c c a t g a a a c g g a c a g c c g a c g g a a g c g a g t t c g a t g c t g a g t g a t a t c c c t c t c g g c g g t g g t a c t t t g c c t g t c g g c t g c c t t c g c t c a a g c t

660 130 PKKKRKV (in frame with SV40 NLS) SEVEFSHEYWMR SV40 NLS SPKKKRKVSEVEFSHEYWMR *WLLLPFDRFDLKAVLIPHP g t c a c c a a a g a a g a a g c g g a a a g t c t c t g a a g t c g a g t t t a g c c a c g a g t a t t g g a t g a g c a g t g g t t t c t t c t t c g c c t t t c a g a g a c t t c a g c t c a a a t c g g t g c t c a t a a c c t a c t c

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

780 (in frame with SV40 NLS) LVHNNRVIGEGWNRPIGRHD LVHNNRVIGEGWNRPIGRHD QHVVISHDSLSPVPWDAAVV g c t g g t g c a c a a c a a t a g a g t g a t c g g a g a g g g a t g g a a c a g g c c a a t c g g c c g c c a c g a c g a c c a c g t g t t g t t a t c t c a c t a g c c t c t c c c t a c c t t g t c c g g t t a g c c g g c g g t g c t

StuI* 840 (in frame with SV40 NLS) PTAHAEIMALRQGGLVMQNY PTAHAEIMALRQGGLVMQNY * RGCVCLDHCQPLSAQDHLIV c c c t a c c g c a c a c g c a g a g a t c a t g g c a c t g a g g c a g g g a g g c c t g g t c a t g c a g a a t t a g g g a t g g c g t g t g c g t c t c t a g t a c c g t g a c t c c g t c c c t c c g g a c c a g t a c g t c t t a a t

BspDI*ClaI* 900 (in frame with SV40 NLS) RLIDATLYVTLEPCVMCAGA RLIDATLYVTLEPCVMCAGA RRISAVRYTVSSGHTIHAPA AQDIGGQIHCQLWAHHACSC c c g c c t g a t c g a t g c c a c c c t g t a t g t g a c a c t g g a g c c a t g c g t g a t g t g c g c a g g a g c g g c g g a c t a g c t a c g g t g g g a c a t a c a c t g t g a c c t c g g t a c g c a c t a c a c g c g t c c t c g

960 (in frame with SV40 NLS) MIHSRIGRVVFGARDAKTGA MIHSRIGRVVFGARDAKTGA IIWLLIPLTTNPARSALVPA HDVAPDSSHHESCPVGLGAC a a t g a t c c a c a g c a g g a t c g g a a g a g t g g t g t t c g g a g c a c g g g a c g c c a a g a c c g g c g c t t a c t a g g t g t c g t c c t a g c c t t c t c a c c a c a a g c c t c g t g c c c t g c g g t t c t g g c c g c g

1020 (in frame with SV40 NLS) AGSLMDVLHHPGMNHRVEIT AGSLMDVLHHPGMNHRVEIT APERISTSCWGPMFWRTSIV CAGQHIHQVVGAHVVPHLDC a g c a g g c t c c c t g a t g g a t g t g c t g c a c c a c c c c g g c a t g a a c c a c c g g g t g g a g a t c a c t c g t c c g a g g g a c t a c c t a c a c g a c g t g g t g g g g c c g t a c t t g g t g g c c c a c c t c t a g t g

1080 (in frame with SV40 NLS) EGILADECAALLSDFFRMRR EGILADECAALLSDFFRMRR SPIRASSHAARSLSKKLIRL LSDQCVLAGGQQAIEKSHPS a g a g g g a a t c c t g g c a g a c g a g t g c g c c g c c c t g c t g a g c g a t t t c t t t a g a a t g c g g a g t c t c c c t t a g g a c c g t c t g c t c a c g c g g c g g g a c g a c t c g c t a a a g a a a t c t t a c g c c t c

1140 (in frame with SV40 NLS) QEIKAQKKAQSSTDSGGSSG QEIKAQKKAQSSTDSGGSSG CSILAWFFACLEVSEPPDLP LLDLGLLLCLAGGVRSSRAS a c a g g a g a t c a a g g c c c a g a a g a a g g c a c a g a g c t c c a c c g a c t c t g g a g g a t c t a g c g g t g t c c t c t a g t t c c g g g t c t t c t t c c g t g t c t c g a g g t g g c t g a g a c c t c c t a g a t c g c c

1200 (in frame with SV40 NLS) GSSGSETPGTSESATPESSG GSSGSETPGTSESATPESSG PDEPLSVGPVLSDAVGSLEP SGRSALCWACALGGCWLAGA a g g a t c c t c t g g a a g c g a g a c a c c a g g c a c a a g c g a g t c c g c c a c a c c a g a g a g c t c c g g t c c t a g g a g a c c t t c g c t c t g t g g t c c g t g t t c g c t c a g g c g g t g t g g t c t c t c g a g g c c

1260 (in frame with SV40 NLS) GSSGGSSEVEFSHEYWMRHA GSSGGSSEVEFSHEYWMRHA PEEPPDESTSNEWSYQILCA AGGSSGRLHLKGVLVPHSMG c g g c t c c t c c g g a g g a t c c t c t g a g g t g g a g t t t t c c c a c g a g t a c t g g a t g a g a c a t g c g c c g a g g a g g c c t c c t a g g a g a c t c c a c c t c a a a a g g g t g c t c a t g a c c t a c t c t g t a c g

1320 (in frame with SV40 NLS) LTLAKRARDEREVPVGAVLV LTLAKRARDEREVPVGAVLV RVRALLARSSLSTGTPATST QGQGLPCAILPLHRHSGHQH c c t g a c c c t g g c c a a g a g g g c a c g c g a t g a g a g g g a g g t g c c t g t g g g a g c c g t g c t g g t g g a c t g g g a c c g g t t c t c c c g t g c g c t a c t c t c c c t c c a c g g a c a c c c t c g g c a c g a c c a

1380 (in frame with SV40 NLS) LNNRVIGEGWNRAIGLHDPT LNNRVIGEGWNRAIGLHDPT SFLLTIPSPQFLAMPRCSGV QVISHDALAPVSGDAQVVWC g c t g a a c a a t a g a g t g a t c g g c g a g g g c t g g a a c a g a g c c a t c g g c c t g c a c g a c c c a a c c g a c t t g t t a t c t c a c t a g c c g c t c c c g a c c t t g t c t c g g t a g c c g g a c g t g c t g g g t t g

1440 (in frame with SV40 NLS) AHAEIMALRQGGLVMQNYRL AHAEIMALRQGGLVMQNYRL AWASIIARLCPPRTM GM a g c c c a t g c c g a a a t t a t g g c c c t g a g a c a g g g c g g c c t g g t c a t g c a g a a c t a c a g a c t t c g g g t a c g g c t t t a a t a c c g g g a c t c t g t c c c g c c g g a c c a g t a c g t c t t g a t g t c t g a

MreI 1500 (in frame with SV40 NLS) IDATLYVTFEPCVMCAGAMI IDATLYVTFEPCVMCAGAMI g a t t g a c g c c a c c c t g t a c g t g a c a t t c g a g c c t t g c g t g a t g t g c g c c g g c g c c a t g a t c t a a c t g c g g t g g g a c a t g c a c t g t a a g c t c g g a a c g c a c t a c a c g c g g c c g c g g t a c t a

1560 (in frame with SV40 NLS) HSRIGRVVFGVRNAKTGAAG HSRIGRVVFGVRNAKTGAAG c c a c t c t a g g a t c g g c c g c g t g g t g t t t g g c g t g a g g a a c g c a a a a a c c g g c g c c g c a g g g g t g a g a t c c t a g c c g g c g c a c c a c a a a c c g c a c t c c t t g c g t t t t t g g c c g c g g c g t c c

1620 (in frame with SV40 NLS) SLMDVLHYPGMNHRVEITEG SLMDVLHYPGMNHRVEITEG c t c c c t g a t g g a c g t g c t g c a c t a c c c c g g c a t g a a t c a c c g c g t c g a a a t t a c c g a g g g g a g g g a c t a c c t g c a c g a c g t g a t g g g g c c g t a c t t a g t g g c g c a g c t t t a a t g g c t c c c

1680 (in frame with SV40 NLS) ILADECAALLCYFFRMPRQV ILADECAALLCYFFRMPRQV a a t c c t g g c a g a t g a a t g t g c c g c c c t g c t g t g c t a t t t c t t t c g g a t g c c t a g a c a g g t t t a g g a c c g t c t a c t t a c a c g g c g g g a c g a c a c g a t a a a g a a a g c c t a c g g a t c t g t c c a

1740 (in frame with SV40 NLS) FNAQKKAQSSTDSGGSSGGS FNAQKKAQSSTDSGGSSGGS g t t c a a t g c t c a g a a g a a g g c c c a g a g c t c c a c c g a c t c c g g a g g a t c t a g c g g a g g c t c c a a g t t a c g a g t c t t c t t c c g g g t c t c g a g g t g g c t g a g g c c t c c t a g a t c g c c t c c g a g

1800 (in frame with SV40 NLS) SGSETPGTSESATPESSGGS SGSETPGTSESATPESSGGS c t c t g g c t c t g a g a c a c c t g g c a c a a g c g a g a g c g c a a c a c c t g a a a g c a g c g g g g g c a g g a g a c c g a g a c t c t g t g g a c c g t g t t c g c t c t c g c g t t g t g g a c t t t c g t c g c c c c c g t c

1860 SGGS 5 10 15 DKKYSIGLAIGTNSVG Cas9 5 DKKYSIGL 10 A 15 IGTNSVG Cas9(D10A) SGGSDKKYSIGLAIGTNSVG *VLLVADAQGDAGVRHA c a g c g g g g g g t c a g a c a a g a a g t a c a g c a t c g g c c t g g c c a t c g g c a c c a a c t c t g t g g g g t c g c c c c c c a g t c t g t t c t t c a t g t c g t a g c c g g a c c g g t a g c c g t g g t t g a g a c a c c c

1920 20 25 30 35 WAVITDEYKVPSKKFKVLGN Cas9(D10A) 20 25 30 35 WAVITDEYKVPSKKFKVLGN Cas9 WAVITDEYKVPSKKFKVLGN PGHDGVLVLHGALFELHQAV 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 t t c a a g g t g c t g g g c a a 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 a a g t t c c a c g a c c c g t t

1980 40 45 50 55 TDRHSIKKNLIGALLFDSGE Cas9(D10A) 40 45 50 55 TDRHSIKKNLIGALLFDSGE Cas9 TDRHSIKKNLIGALLFDSGE GVPVADLLVQDSGQQEVAAF 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 c t g t t c g a c a g c g g c g a 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 g a c a a g c t g t c g c c g c t

2040 60 65 70 75 TAEATRLKRTARRRYTRRKN Cas9(D10A) 60 65 70 75 TAEATRLKRTARRRYTRRKN Cas9 TAEATRLKRTARRRYTRRKN CGLGGPQLSGGSSSVGSPLV 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 t a c a c c a g a c g g a a g a a 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 a t g t g g t c t g c c t t c t t

BglII 2100 80 85 90 95 RICYLQEIFSNEMAKVDDSF Cas9(D10A) 80 85 90 95 RICYLQEIFSNEMAKVDDSF Cas9 RICYLQEIFSNEMAKVDDSF PDAIQLLDEAVLHGLHVVAE 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 a g g t g g a c g a c a g 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 t t c c a c c t g c t g t c g a a

2160 100 105 110 115 FHRLEESFLVEEDKKHERHP Cas9(D10A) 100 105 110 115 FHRLEESFLVEEDKKHERHP Cas9 FHRLEESFLVEEDKKHERHP EVSQFLGEQHFLILLVLPVG 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 a a g c a c g a g c g g c a c c c 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 t t c g t g c t c g c c g t g g g

2220 120 125 130 135 IFGNIVDEVAYHEKYPTIYH Cas9(D10A) 120 125 130 135 IFGNIVDEVAYHEKYPTIYH Cas9 IFGNIVDEVAYHEKYPTIYH DEAVDHVLHGVVLLVGGDVV 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 a c c c c a c c a t c t a c c a 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 a t g g g g t g g t a g a t g g t

2280 140 145 150 155 LRKKLVDSTDKADLRLIYLA Cas9(D10A) 140 145 150 155 LRKKLVDSTDKADLRLIYLA Cas9 LRKKLVDSTDKADLRLIYLA QSLFQHVAGVLGVQPQDIQG 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 c g g c t g a t c t a t c t g g c 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 g c c g a c t a g a t a g a c c g

2340 160 165 170 175 LAHMIKFRGHFLIEGDLNPD Cas9(D10A) 160 165 170 175 LAHMIKFRGHFLIEGDLNPD Cas9 LAHMIKFRGHFLIEGDLNPD QGVHDLEPAVEQDLAVQVGV 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 g c g a c c t g a a c c c c g a 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 c c g c t g g a c t t g g g g c t

2400 180 185 190 195 NSDVDKLFIQLVQTYNQLFE Cas9(D10A) 180 185 190 195 NSDVDKLFIQLVQTYNQLFE Cas9 NSDVDKLFIQLVQTYNQLFE VAVHVLQEDLQHLGVVLQEL 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 t a c a a c c a g c t g t t c g a 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 a t g t t g g t c g a c a a g c t

2460 200 205 210 215 ENPINASGVDAKAILSARLS Cas9(D10A) 200 205 210 215 ENPINASGVDAKAILSARLS Cas9 ENPINASGVDAKAILSARLS FVGDVGAAHVGLGDQRGSQA 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 c t g t c t g c c a g a c t g a g 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 g a c a g a c g g t c t g a c t c

2520 220 225 230 235 KSRRLENLIAQLPGEKKNGL Cas9(D10A) 220 225 230 235 KSRRLENLIAQLPGEKKNGL Cas9 KSRRLENLIAQLPGEKKNGL LASPQFIQDGLQGALLLIAQ 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 g a a g a a g a a t g g c c t g t t c t c g t c t g c c g a c c t t t t a g a c t a g c g g g t c g a c g g g c c g c t c t t c t t c t t a c c g g a

2580 240 245 250 255 FGNLIALSLGLTPNFKSNFD Cas9(D10A) 240 245 250 255 FGNLIALSLGLTPNFKSNFD Cas9 FGNLIALSLGLTPNFKSNFD ESVQNGQAQAQGGVELAVEV 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 t t c a a g a g c a a c t t c 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 a a g t t c t c g t t g a a g c t

2640 260 265 270 275 LAEDAKLQLSKDTYDDDLDN Cas9(D10A) 260 265 270 275 LAEDAKLQLSKDTYDDDLDN Cas9 LAEDAKLQLSKDTYDDDLDN QGLIGFQLQALVGVVVVQVV 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 g a c g a c g a c c t g g a c a a 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 c t g c t g c t g g a c c t g t t

2700 280 285 290 295 LLAQIGDQYADLFLAAKNLS Cas9(D10A) 280 285 290 295 LLAQIGDQYADLFLAAKNLS Cas9 LLAQIGDQYADLFLAAKNLS QQGLDAVLVGVQKQGGLVQG 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 g c c g c c a a g a a c c t g t c 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 c g g c g g t t c t t g g a c a g

2760 300 305 310 315 DAILLSDILRVNTEITKAPL Cas9(D10A) 300 305 310 315 DAILLSDILRVNTEITKAPL Cas9 DAILLSDILRVNTEITKAPL VGDQQAVDQSHVGLDGLGGQ 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 a t c a c c a a g g c c c c c c t 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 t a g t g g t t c c g g g g g g a

2820 320 325 330 335 SASMIKRYDEHHQDLTLLKA Cas9(D10A) 320 325 330 335 SASMIKRYDEHHQDLTLLKA Cas9 SASMIKRYDEHHQDLTLLKA AGRHDLSVVLVVLVQGQQFS 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 c t g a c c c t g c t g a a a g c 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 g a c t g g g a c g a c t t t c g

2880 340 345 350 355 LVRQQLPEKYKEIFFDQSKN Cas9(D10A) 340 345 350 355 LVRQQLPEKYKEIFFDQSKN Cas9 LVRQQLPEKYKEIFFDQSKN EHPLLQRLLVFLNEEVLALV 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 t t c g a c c a g a g c a a g a a 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 a a g c t g g t c t c g t t c t t

2940 360 365 370 375 GYAGYIDGGASQEEFYKFIK Cas9(D10A) 360 365 370 375 GYAGYIDGGASQEEFYKFIK Cas9 GYAGYIDGGASQEEFYKFIK AVGAVNVASGALFLEVLEDL 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 t t c t a c a a g t t c a t c a a 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 a a g a t g t t c a a g t a g t t

3000 380 385 390 395 PILEKMDGTEELLVKLNRED Cas9(D10A) 380 385 390 395 PILEKMDGTEELLVKLNRED Cas9 PILEKMDGTEELLVKLNRED GDQFLHVAGLFQEHLQVSLV 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 c t g a a c a g a g a g g a 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 t t c g a c t t g t c t c t c c t

3060 400 405 410 415 LLRKQRTFDNGSIPHQIHLG Cas9(D10A) 400 405 410 415 LLRKQRTFDNGSIPHQIHLG Cas9 LLRKQRTFDNGSIPHQIHLG QQPLLPGEVVAADGVLDVQS 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 c a c c a g a t c c a c c t g g g 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 g t g g t c t a g g t g g a c c c

3120 420 425 430 435 ELHAILRRQEDFYPFLKDNR Cas9(D10A) 420 425 430 435 ELHAILRRQEDFYPFLKDNR Cas9 ELHAILRRQEDFYPFLKDNR LQVGNQPPLFIKVWEQLVVP 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 t t c c t g a a g g a c a a c c g 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 a a g g a c t t c c t g t t g g c

PspOMIApaI 3180 440 445 450 455 EKIEKILTFRIPYYVGPLAR Cas9(D10A) 440 445 450 455 EKIEKILTFRIPYYVGPLAR Cas9 EKIEKILTFRIPYYVGPLAR FLDLLDQGEADGVVHARQGP 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 g t g g g c c c t c t g g c c a g 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 c a c c c g g g a g a c c g g t c

3240 460 465 470 475 GNSRFAWMTRKSEETITPWN Cas9(D10A) 460 465 470 475 GNSRFAWMTRKSEETITPWN Cas9 GNSRFAWMTRKSEETITPWN SVASEGPHGSLALFGDGGPV 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 a c c a t c a c c c c c t g g a a 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 t g g t a g t g g g g g a c c t t

3300 480 485 490 495 FEEVVDKGASAQSFIERMTN Cas9(D10A) 480 485 490 495 FEEVVDKGASAQSFIERMTN Cas9 FEEVVDKGASAQSFIERMTN ELFHHVLASGGLAEDLPHGV 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 a t c g a g c g g a t g a c c a a 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 t a g c t c g c c t a c t g g t t

3360 500 505 510 515 FDKNLPNEKVLPKHSLLYEY Cas9(D10A) 500 505 510 515 FDKNLPNEKVLPKHSLLYEY Cas9 FDKNLPNEKVLPKHSLLYEY EILVQGVLLHQGLVAQQVLV 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 a g c c t g c t g t a c g a g t a 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 t c g g a c g a c a t g c t c a t

3420 520 525 530 535 FTVYNELTKVKYVTEGMRKP Cas9(D10A) 520 525 530 535 FTVYNELTKVKYVTEGMRKP Cas9 FTVYNELTKVKYVTEGMRKP EGHIVLQGFHFVHGLSHSLG 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 g a g g g a a t g a g a a a g c c 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 c t c c c t t a c t c t t t c g g

3480 540 545 550 555 AFLSGEQKKAIVDLLFKTNR Cas9(D10A) 540 545 550 555 AFLSGEQKKAIVDLLFKTNR Cas9 AFLSGEQKKAIVDLLFKTNR GEQAALLFLGDHVQQELGVP 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 t g c t g t t c a a g a c c a a c c 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 a c g a c a a g t t c t g g t t g g c

3540 560 565 570 575 KVTVKQLKEDYFKKIECFDS Cas9(D10A) 560 565 570 575 KVTVKQLKEDYFKKIECFDS Cas9 KVTVKQLKEDYFKKIECFDS FHGHLLQFLVVELFDLAEVG 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 a t c g a g t g c t t c g a c t 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 t a g c t c a c g a a g c t g a g

PasI 3600 580 585 590 595 VEISGVEDRFNASLGTYHDL Cas9(D10A) 580 585 590 595 VEISGVEDRFNASLGTYHDL Cas9 VEISGVEDRFNASLGTYHDL HFDGAHFIPEVGGQACVVIQ 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 g g c a c a t a c c a c g a t c t 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 c c g t g t a t g g t g c t a g a

3660 600 605 610 615 LKIIKDKDFLDNEENEDILE Cas9(D10A) 600 605 610 615 LKIIKDKDFLDNEENEDILE Cas9 LKIIKDKDFLDNEENEDILE QFNDLVLVEQVILFVLVNQF 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 a a c g a g g a c a t t c t g 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 t t g c t c c t g t a a g a c c t

EcoRV 3720 620 625 630 635 DIVLTLTLFEDREMIEERLK Cas9(D10A) 620 625 630 635 DIVLTLTLFEDREMIEERLK Cas9 DIVLTLTLFEDREMIEERLK IDHQGQCQKLVSLHDLFPQF 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 a t c g a g g a a c g g c t g a a 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 t a g c t c c t t g c c g a c t t

3780 640 645 650 655 TYAHLFDDKVMKQLKRRRYT Cas9(D10A) 640 645 650 655 TYAHLFDDKVMKQLKRRRYT Cas9 TYAHLFDDKVMKQLKRRRYT *AWRNSSLTIFCSFRRLYV GIGVQEVVFHHLLQLPPSVG 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 a a g c g g c g g a g a t a c a c 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 t t c g c c g c c t c t a t g t g

3840 660 665 670 675 GWGRLSRKLINGIRDKQSGK Cas9(D10A) 660 665 670 675 GWGRLSRKLINGIRDKQSGK Cas9 GWGRLSRKLINGIRDKQSGK PQPLSLRFSILPMRSLCDPL APAPQAPLQDVADPVLLGAL 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 g a c a a g c a g t c c g g c a a 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 c t g t t c g t c a g g c c g t t

3900 680 685 690 695 TILDFLKSDGFANRNFMQLI Cas9(D10A) 680 685 690 695 TILDFLKSDGFANRNFMQLI Cas9 TILDFLKSDGFANRNFMQLI VIRSKRFDSPKALLFKMCSI CDQIEQLGVAEGVSVEHLQD 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 a a c t t c a t g c a g c t g a 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 t t g a a g t a c g t c g a c t a

3960 700 705 710 715 HDDSLTFKEDIQKAQVSGQG Cas9(D10A) 700 705 710 715 HDDSLTFKEDIQKAQVSGQG Cas9 HDDSLTFKEDIQKAQVSGQG WSSLRVKLSSMWFAWTDPWP VVVAQGKFLVDLFGLHGALA 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 c a g g t g t c c g g c c a g g g 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 g t c c a c a g g c c g g t c c c

4020 720 725 730 735 DSLHEHIANLAGSPAIKKGI Cas9(D10A) 720 725 730 735 DSLHEHIANLAGSPAIKKGI Cas9 DSLHEHIANLAGSPAIKKGI SLRCSCMALRAPLGAMLFPM IAQVLVNGIQGAAGGNLLAD 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 c c a t t a a g a a g g g c a t 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 c g g t a a t t c t t c c c g t a

4080 740 745 750 755 LQTVKVVDELVKVMGRHKPE Cas9(D10A) 740 745 750 755 LQTVKVVDELVKVMGRHKPE Cas9 LQTVKVVDELVKVMGRHKPE RCVTFTTSSSTFTIPRCLGS QLCHLHHVLEHFHHAPVLGL 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 g g c c g g c a c a a g c c c g 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 c c g g c c g t g t t c g g g c t

4140 760 765 770 775 NIVIEMARENQTTQKGQKNS Cas9(D10A) 760 765 770 775 NIVIEMARENQTTQKGQKNS Cas9 NIVIEMARENQTTQKGQKNS FMTISIALSFWVVWFPCFFL VDHDFHGSLVLGGLLSLLVA 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 g a a g g g a c a g a a g a a c a g 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 t c t t c c c t g t c t t c t t g t c

4200 780 785 790 795 RERMKRIEEGIKELGSQILK Cas9(D10A) 780 785 790 795 RERMKRIEEGIKELGSQILK Cas9 RERMKRIEEGIKELGSQILK RSLIFRISSPM ALSHLPDFLADFLQAALDQF 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 g g c a g c c a g a t c c t g a a 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 c c g t c g g t c t a g g a c t t

4260 800 805 810 815 EHPVENTQLQNEKLYLYYLQ Cas9(D10A) 800 805 810 815 EHPVENTQLQNEKLYLYYLQ Cas9 EHPVENTQLQNEKLYLYYLQ FVGHFVGLQLVLLQVQVVQL 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 t a c c t g t a c t a c c t g c a 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 a t g g a c a t g a t g g a c g t

4320 820 825 830 835 NGRDMYVDQELDINRLSDYD Cas9(D10A) 820 825 830 835 NGRDMYVDQELDINRLSDYD Cas9 NGRDMYVDQELDINRLSDYD IPPIHVHVLFQVDVPQGVVI 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 c g g c t g t c c g a c t a c g a 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 g c c g a c a g g c t g a t g c t

4380 840 845 850 855 VDHIVPQSFLKDDSIDNKVL Cas9(D10A) 840 845 850 855 VDHIVPQSFLKDDSIDNKVL Cas9 VDHIVPQSFLKDDSIDNKVL HVM 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 a t c g a c a a c a a g g 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 t a g c t g t t g t t c c a c g a

4440 860 865 870 875 TRSDKNRGKSDNVPSEEVVK Cas9(D10A) 860 865 870 875 TRSDKNRGKSDNVPSEEVVK Cas9 TRSDKNRGKSDNVPSEEVVK 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 c c g a a g a g g t c g t g a 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 a g g c t t c t c c a g c a c t t

4500 880 885 890 895 KMKNYWRQLLNAKLITQRKF Cas9(D10A) 880 885 890 895 KMKNYWRQLLNAKLITQRKF Cas9 KMKNYWRQLLNAKLITQRKF 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 t g a t t a c c c a g a g a a a g 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 a c t a a t g g g t c t c t t t c a a

4560 900 905 910 915 DNLTKAERGGLSELDKAGFI Cas9(D10A) 900 905 910 915 DNLTKAERGGLSELDKAGFI Cas9 DNLTKAERGGLSELDKAGFI 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 g a t a a g g c c g g c t t c a t 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 c t a t t c c g g c c g a a g t a

PmlI 4620 920 925 930 935 KRQLVETRQITKHVAQILDS Cas9(D10A) 920 925 930 935 KRQLVETRQITKHVAQILDS Cas9 KRQLVETRQITKHVAQILDS 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 g c a c a g a t c c t g g a c t c 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 c g t g t c t a g g a c c t g a g

4680 940 945 950 955 RMNTKYDENDKLIREVKVIT Cas9(D10A) 940 945 950 955 RMNTKYDENDKLIREVKVIT Cas9 RMNTKYDENDKLIREVKVIT 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 g a a g t g a a a g t g a t c a c 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 c t t c a c t t t c a c t a g t g

4740 960 965 970 975 LKSKLVSDFRKDFQFYKVRE Cas9(D10A) 960 965 970 975 LKSKLVSDFRKDFQFYKVRE Cas9 LKSKLVSDFRKDFQFYKVRE 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 t t t t a c a a a g t g c g c g a 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 a a a a t g t t t c a c g c g c t

4800 980 985 990 995 INNYHHAHDAYLNAVVGTAL Cas9(D10A) 980 985 990 995 INNYHHAHDAYLNAVVGTAL Cas9 INNYHHAHDAYLNAVVGTAL g a t c a a c a a c t a c c a c c a c g c c c a c g a c g c c t a c c t g a a c g c c g t c g t g g g a a c c g c c c t c t a g t t g t t g a t g g t g g t g c g g g t g c t g c g g a t g g a c t t g c g g c a g c a c c c t t g g c g g g a

4860 1000 1005 1010 1015 IKKYPKLESEFVYGDYKVYD Cas9(D10A) 1000 1005 1010 1015 IKKYPKLESEFVYGDYKVYD Cas9 IKKYPKLESEFVYGDYKVYD g a t c a a a a a g t a c c c t a a g c t g g a a a g c g a g t t c g t g t a c g g c g a c t a c a a g g t g t a c g a c t a g t t t t t c a t g g g a t t c g a c c t t t c g c t c a a g c a c a t g c c g c t g a t g t t c c a c a t g c t

4920 1020 1025 1030 1035 VRKMIAKSEQEIGKATAKYF Cas9(D10A) 1020 1025 1030 1035 VRKMIAKSEQEIGKATAKYF Cas9 VRKMIAKSEQEIGKATAKYF c g t g c g g a a g a t g a t c g c c a a g a g c g a g c a g g a a 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 g c a c g c c t t c t a c t a g c g g t t c t c g c t c g t c c t t 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

4980 1040 1045 1050 1055 FYSNIMNFFKTEITLANGEI Cas9(D10A) 1040 1045 1050 1055 FYSNIMNFFKTEITLANGEI Cas9 FYSNIMNFFKTEITLANGEI c t t c t a c a g c a a c a t c a t g a a c t t t t t c a a g a c c 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 g a a g a t g t c g t t g t a g t a c t t g a a a a a g t t c t g g 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

5040 1060 1065 1070 1075 RKRPLIETNGETGEIVWDKG Cas9(D10A) 1060 1065 1070 1075 RKRPLIETNGETGEIVWDKG Cas9 RKRPLIETNGETGEIVWDKG c c g g a a g c g g c c t c t g a t c g a g a c a a a c g g c g a a 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 g g c c t t c g c c g g a g a c t a g c t c t g t t t g c c g c t t 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

5100 1080 1085 1090 1095 RDFATVRKVLSMPQVNIVKK Cas9(D10A) 1080 1085 1090 1095 RDFATVRKVLSMPQVNIVKK Cas9 RDFATVRKVLSMPQVNIVKK c c g g g a t t t t g c c a c c g t g c g g a a a g t g c t g a g c 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 g c c c t a a a a c g g t g g c a c g c c t t t c a c g a c t c g 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

5160 1100 1105 1110 1115 TEVQTGGFSKESILPKRNSD Cas9(D10A) 1100 1105 1110 1115 TEVQTGGFSKESILPKRNSD Cas9 TEVQTGGFSKESILPKRNSD g a c c g a g g t g c a g a c a g g c g g c t t c a g c a a a g a g t 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 c t g g c t c c a c g t c t g t c c g c c g a a g t c g t t t c t c 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

KflI 5220 1120 1125 1130 1135 KLIARKKDWDPKKYGGFDSP Cas9(D10A) 1120 1125 1130 1135 KLIARKKDWDPKKYGGFDSP Cas9 KLIARKKDWDPKKYGGFDSP t a a g c t g a t c g c c a g a a a g a a g g a c t g g g a c c c t a 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 a t t c g a c t a g c g g t c t t t c t t c c t g a c c c t g g g a 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

5280 1140 1145 1150 1155 TVAYSVLVVAKVEKGKSKKL Cas9(D10A) 1140 1145 1150 1155 TVAYSVLVVAKVEKGKSKKL Cas9 TVAYSVLVVAKVEKGKSKKL c a c c g t g g c c t a t t c t g t g c t g g t g g t g g c c a a a 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 t g g c a c c g g a t a a g a c a c g a c c a c c a c c g g t t t 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

BsaBI* 5340 1160 1165 1170 1175 KSVKELLGITIMERSSFEKN Cas9(D10A) 1160 1165 1170 1175 KSVKELLGITIMERSSFEKN Cas9 KSVKELLGITIMERSSFEKN g a a g a g t g t g a a a g a g c t g c t g g g g a t c a c c a t c 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 c t t c t c a c a c t t t c t c g a c g a c c c c t a g t g g t a g 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

5400 1180 1185 1190 1195 PIDFLEAKGYKEVKKDLIIK Cas9(D10A) 1180 1185 1190 1195 PIDFLEAKGYKEVKKDLIIK Cas9 PIDFLEAKGYKEVKKDLIIK t c c c a t c g a c t t t c t g g a a g c c a a g g g c t a c a a a 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 a g g g t a g c t g a a a g a c c t t c g g t t c c c g a t g t t t 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

5460 1200 1205 1210 1215 LPKYSLFELENGRKRMLASA Cas9(D10A) 1200 1205 1210 1215 LPKYSLFELENGRKRMLASA Cas9 LPKYSLFELENGRKRMLASA g c t g c c t a a g t a c t c c c t g t t c g a g c t g g a a a a c 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

5520 1220 1225 1230 1235 GELQKGNELALPSKYVNFLY Cas9(D10A) 1220 1225 1230 1235 GELQKGNELALPSKYVNFLY Cas9 GELQKGNELALPSKYVNFLY c g g c g a a c t g c a g a a g g g a a a c g a a c t g g c c c t g c c c t c c a a a t a t g t g a a c t t c c t g t a g c c g c t t g a c g t c t t c c c t t t g c t t g a c c g g g a 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

5580 1240 1245 1250 1255 LASHYEKLKGSPEDNEQKQL Cas9(D10A) 1240 1245 1250 1255 LASHYEKLKGSPEDNEQKQL Cas9 LASHYEKLKGSPEDNEQKQL c c t g g c c a g c c a c t a t g a g a a g c t g a a g g g c t c c 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 g a c c g g t c g g t g a t a c t c t t c g a c t t c c c g a g g 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

5640 1260 1265 1270 1275 FVEQHKHYLDEIIEQISEFS Cas9(D10A) 1260 1265 1270 1275 FVEQHKHYLDEIIEQISEFS Cas9 FVEQHKHYLDEIIEQISEFS g t t t g t g g a a c a g c a c a a g c a c t a c c t g g a c g a g 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

5700 1280 1285 1290 1295 KRVILADANLDKVLSAYNKH Cas9(D10A) 1280 1285 1290 1295 KRVILADANLDKVLSAYNKH Cas9 KRVILADANLDKVLSAYNKH c a a g a g a g t g a t c c t g g c c g a c g c t a a t c t g g a c 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 g t t c t c t c a c t a g g a c c g g c t g c g a t t a g a c c t g 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

5760 1300 1305 1310 1315 RDKPIREQAENIIHLFTLTN Cas9(D10A) 1300 1305 1310 1315 RDKPIREQAENIIHLFTLTN Cas9 RDKPIREQAENIIHLFTLTN c c g g g a t a a g c c c a t c a g a g a g c a g g c c g a g a a t 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 g g c c c t a t t c g g g t a g t c t c t c g t c c g g c t c t t a 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

5820 1320 1325 1330 1335 LGAPAAFKYFDTTIDRKRYT Cas9(D10A) 1320 1325 1330 1335 LGAPAAFKYFDTTIDRKRYT Cas9 LGAPAAFKYFDTTIDRKRYT t c t g g g a g c c c c t g c c g c c t t c a a g t a c t t t g a 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 a g a c c c t c g g g g a c g g c g g a a g t t c a t g a a a c t 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

5880 1340 1345 1350 1355 STKEVLDATLIHQSITGLYE Cas9(D10A) 1340 1345 1350 1355 STKEVLDATLIHQSITGLYE Cas9 STKEVLDATLIHQSITGLYE c a g c a c c a a a g a g g t g c t g g a c g c c a c c c t g a t 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 t c g t g g t t t c t c c a c g a c c t g c g g t g g g a c t a g 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

5940 1360 1365 TRIDLSQLGGD 1 5 KRPAATKKA Cas9 nucleoplasmin NLS 1360 1365 TRIDLSQLGGD (in frame with Cas9(D10A)) KRPAATKKA Cas9(D10A) TRIDLSQLGGDKRPAATKKA g a c a c g g a t c g a c c t g t c t c a g c t g g g a g g c g a 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 t g t g c c t a g c t g g a c a g a g t c g a c c c t c c g c t 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

FseI 6000 (in frame with Cas9(D10A)) GQAKKKKGS* 10 15 GQAKKKK GS* nucleoplasmin NLS GQAKKKKGS* c g g c c a g g c a a a a a a g a a a a a g g g a t c c t a a g c t c g c t t t c t t g c t g t c c a a t t t c t a t t g c c g g t c c g t t t t t t c t t t t t c c c t a g g a t t c g a g c g a a a g a a c g a c a g g t t a a a g a t a a

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

BmtINheI 6120 a g c a t c t g g a t t c t g c c t a a t a a a a a a c a t t t a t t t t c a t t g c t a g c t c g c t t t c t t g c t t c g t a g a c c t a a g a c g g a t t a t t t t t t g t a a a t a a a a g t a a c g a t c g a g c g a a a g a a c g a

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

EcoRI 6240 t a t g a a g g g c c t t g a g c a t c t g g a t t c t g c c t a a t a a a a a a c a t t t a t t t t c a t t g c g a a a t a c t t c c c g g a a c t c g t a g a c c t a a g a c g g a t t a t t t t t t g t a a a t a a a a g t a a c g c t t

XbaI 6300 bGH poly(A) signal G G A G C T G A C A C G G A A G A T BGH-rev t t c t a g a a 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 t t c t a g t t g c c a g c c a a a g a t c t t 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 a a g a t c a a c g g t c g g t

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

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

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

6540 bGH poly(A) signal AAV2 ITR AAV2 ITR g g g g a g g t a 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 c t c t g c g c g c t c c c c t c c a t 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 g a g a g a c g c g c g a

6600 AAV2 ITR AAV2 ITR 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 c t t t g c c c g g g c g 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 g a a a c g g g c c c g c

SbfI 6660 AAV2 ITR AAV2 ITR 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 c g g t a t t t t c t 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 c g c c a t a a a a g a g

6720 f1 ori 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 t a g t a c g c g c c c t 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 a t c a t g c g c g g g a

6780 f1 ori 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 c c g c t a c a c t t g 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 t g g c g a t g t g a a c

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

6900 f1 ori 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 t t t a g t g c t t t a 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 a a a t c a c g a a a t g

6960 f1 ori 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 g g g c c a t c g c c c t 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 c c c g g t a g c g g g a

7020 f1 ori G T G G A C T C T T G T F1ori-F 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 a g t g g a c t c t t g t 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 t c a c c t g a g a a c a

PsiI 7080 f1 ori T C C A A A C T G G F1ori-F t c c a a a c t g g a a c a a c a c t c a a c c c t a t c t c g g g c t a t t c t t t t g a t t t a t a a g g g a t t t a g g t t t g a c c t t g t t g t g a g t t g g g a t a g a g c c c g a t a a g a a a a c t a a a t a t t c c c t a a a

7140 f1 ori t g c c g a t t t c g g c c t a t t g g t t a a a a a a t g a g c t g a t t t a a c a a a a a t t t a a c g c g a a t t a c g g c t a a a g c c g g a t a a c c a a t t t t t t a c t c g a c t a a a t t g t t t t t a a a t t g c g c t t a a

7200 f1 ori A T G T T A G A C G A G A C T A C pRS-marker 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 t c t g c t c t g a t g 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 t a g a c g a g a c t a c

7260 G G C pRS-marker 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 c c c t g a c g g g c t t 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 g g g a c t g c c c g a a

7320 C C G G G A G C T G C A T G T G T C pGEX 3' 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 a g c t g c a t g t g t c 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 t c g a c g t a c a c a g

7380 A G A G G pGEX 3' C G G A G C A C T A T G C G G A T A pBRforEco 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 g t g a t a c g c c t a t 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 c a c t a t g c g g a t a

7440 A pBRforEco 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 g g c a c t t t t c g g g 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 c c g t g a a a a g c c c

7500 AmpR promoter 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 a a t a t g t a t c c g 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 t t a t a c a t a g g c g

7560 1 signal sequ... MS AmpR promoter AmpR MS 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 a a g a g t a t g a g t a 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 t t c t c a t a c t c a t

7620 5 10 15 20 signal sequence IQHFRVALIPFFAAFCLPVF AmpR IQHFRVALIPFFAAFCLPVF 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 c t t c c t g t t t t t g 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 g a a g g a c a a a a a c

7680 A 25 30 35 40 HPETLVKVKDAEDQLGARV AmpR AHPETLVKVKDAEDQLGARV 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 g t g c a c g a g t g g 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 c c a c g t g c t c a c c

7740 45 50 55 60 GYIELDLNSGKILESFRPEE AmpR GYIELDLNSGKILESFRPEE 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 g c c c c g a a g a a 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 g c g g g g c t t c t t g

7800 65 70 75 80 RFPMMSTFKVLLCGAVLSRI AmpR C G A T A C A C C G C G C C A T A A T A Amp-R RFPMMSTFKVLLCGAVLSRI 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 t t a t c c c g t a 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 a a t a g g g c a t a a c

7860 85 90 95 100 DAGQEQLGRRIHYSQNDLVE AmpR DAGQEQLGRRIHYSQNDLVE 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 g a c t t g g t t g a g t 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 c t g a a c c a a c t c a

7920 105 110 115 120 YSPVTEKHLTDGMTVRELCS AmpR YSPVTEKHLTDGMTVRELCS 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 g a a t t a t g c a g t g 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 c t t a a t a c g t c a c

PvuI 7980 125 130 135 140 AAITMSDNTAANLLLTTIGG AmpR AAITMSDNTAANLLLTTIGG 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 a c g a t c g g a g g 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 t g c t a g c c t c c t g

8040 145 150 155 160 PKELTAFLHNMGDHVTRLDR AmpR PKELTAFLHNMGDHVTRLDR 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 c g c c t t g a t c g t t 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 g c g g a a c t a g c a a

8100 165 170 175 180 WEPELNEAIPNDERDTTMPV AmpR WEPELNEAIPNDERDTTMPV 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 a c g a t g c c t g t a g 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 t g c t a c g g a c a t c

8160 185 190 195 200 AMATTLRKLLTGELLTLASR AmpR AMATTLRKLLTGELLTLASR 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 c t a g c t t c c c g g 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 g a t c g a a g g g c c g

8220 205 210 215 220 QQLIDWMEADKVAGPLLRSA AmpR QQLIDWMEADKVAGPLLRSA 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 c t g c g c t c g g c c c 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 g a c g c g a g c c g g g

8280 225 230 235 240 LPAGWFIADKSGAGERGSRG AmpR LPAGWFIADKSGAGERGSRG 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 g a a g c c g c g g t a 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 c c t t c g g c g c c a t

8340 245 250 255 260 IIAALGPDGKPSRIVVIYTT AmpR IIAALGPDGKPSRIVVIYTT 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 a t c t a c a c g a c g g 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 t a g a t g t g c t g c c

8400 265 270 275 280 GSQATMDERNRQIAEIGASL AmpR GSQATMDERNRQIAEIGASL 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 g g t g c c t c a c t g a 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 c c a c g g a g t g a c t

8460 285 IKHW* AmpR IKHW* 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 a t t g a t t t a a a a c 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 t a a c t a a a t t t t g

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

8580 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 a a g a t c a a a g g a 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 t t c t a g t t t c c t a

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

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

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

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

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

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

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

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

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

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

AflIIIPciI 9240 U6 promoter G A G G G C hU6-F 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 c a c a t g t g a g g g c 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 g t g t a c a c t c c c g

9300 U6 promoter C T A T T T C C C A T G A T T hU6-F c t a t t t c c c a t g a t t c c t t c a t a t t t g c a t a t a c g a t a c a a g g c t g t t a g a g a g a t a a t t g a t a a a g g g t a c t a a g g a a g t a t a a a c g t a t a t g c t a t g t t c c g a c a a t c t c t c t a t t a a

9360 U6 promoter g g a a t t a a t t t g a c t g t a a a c a c a a a g a t a t t a g t a c a a a a t a c g t g a c g t a g a a a g t a a c c t t a a t t a a a c t g a c a t t t g t g t t t c t a t a a t c a t g t t t t a t g c a c t g c a t c t t t c a t t

9420 U6 promoter G A C T A T C A T A T G C T T LKO.1 5' t a a t t t c t t g g g t a g t t t g c a g t t t t a a a a t t a t g t t t t a a a a t g g a c t a t c a t a t g c t t a t t a a a g a a c c c a t c a a a c g t c a a a a t t t t a a t a c a a a a t t t t a c c t g a t a g t a t a c g a a

9480 U6 promoter A C C G T LKO.1 5' a c c g t a a c t t g a a a g t a t t t c g a t t t c t t g g c t t t a t a t a t c t t g t g g a a a g g a c g a a a c t g g c a t t g a a c t t t c a t a a a g c t a a a g a a c c g a a a t a t a t a g a a c a c c t t t c c t g c t t t g

PaeR7IPspXIXhoIAbsIAgeI 9540 U6 promoter a c c g g t g t c t t c c t c g a g g a a g a c c c g t t t g a g a g c t a g a a a t a g c a a g t t c a a a t a a g g t g g c c a c a g a a g g a g c t c c t t c t g g g c a a a c t c t c g a t c t t t a t c g t t c a a g t t t a t t c c

AleI 9600 c t a g t c c g t t a t c a a c t t g g c t g a a t g c c t g c g a g c a t c c c a c c c a a g t g g c a c c g a g t c g a t c a g g c a a t a g t t g a a c c g a c t t a c g g a c g c t c g t a g g g t g g g t t c a c c g t g g c t c a g

3ʹ 5ʹ 9613 g g t g c t t t t t t t g c c a c g a a a a a a a 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.