Addgene SpCas9-mSA* Sequencing Result - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: SpCas9-mSA* Sequencing Result


Map View

NsiI (7879) Neo-R (7752 .. 7771) PluTI (7697) SfoI (7695) NarI (7694) KasI (7693) SphI (7295) Neo-F (7142 .. 7161) ApaLI (6615) pBR322ori-F (6442 .. 6461) StuI (6291) MfeI (6173) HpaI (6164) SV40pA-R (6108 .. 6127) EBV-rev (6054 .. 6073) AleI (6009) NotI (5998) pBluescriptSK (5866 .. 5882) XbaI (5877) SK primer SmaI (5861) TspMI - XmaI (5859) DraIII (5764) BstEII (5568) SfiI (5481) EcoRI (5466) BstBI (5455) BstZ17I (5324) SgrAI (5295) AscI - BssHII (5143) PaqCI (4448) NruI (4428) XmnI (4320) FspAI (4138) BsrGI (96) AseI (160) CMV enhancer NdeI (387) SnaBI (493) CMV-F (672 .. 692) T7 (975 .. 994) T7 promoter Kozak sequence c-myc NLS c-myc NLS PaeR7I - PspXI - XhoI (1158) c-myc NLS EcoRV (1217) PvuI (1304) SbfI (1435) XcmI (1539) SalI (1728) BlpI (2136) Bsu36I (2454) BspHI (3127) BmgBI (3698) HindIII (3719) SpCas9-mSA* 7881 bp

Sequence View

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

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

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

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

300 CMV enhancer g c c t g g c t g a c c g c c c a a c g a c c c c c g c c c a t t g a c g t c a a t a a t g a c g t a t g t t c c c a t c g g a c c g a c t g g c g g g t t g c t g g g g g c g g g t a a c t g c a g t t a t t a c t g c a t a c a a g g g t a

360 CMV enhancer a g t a a c g c c a a t a g g g a c t t t c c a t t g a c g t c a a t g g g t g g a g t a t t t a c g g t a a a c t g c t c a t t g c g g t t a t c c c t g a a a g g t a a c t g c a g t t a c c c a c c t c a t a a a t g c c a t t t g a c g

NdeI 420 CMV enhancer 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 c c g c c c c c t a t t g a c g t c a a t g a 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 g g c g g g g g a t a a c t g c a g t t a c t

480 CMV enhancer c g g t a a a t g g c c c g c c t g g c a t t a t g c c c a g t a c a t g a c c t t a c g g g a c t t t c c t a c t t g g c c a t t t a c c g g g c g g a c c g t a a t a c g g g t c a t g t a c t g g a a t g c c c t g a a a g g a t g a a c

SnaBI 540 CMV enhancer CMV promoter 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 t t t t g g c a g t a c 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 a a a a c c g t c a t g t g

600 CMV promoter 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 c a c c c c a t t g a c g t 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 g t g g g g t a a c t g c a

660 CMV promoter 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 t g t c g t a a t a a c c c 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 a c a g c a t t a t t g g g

720 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 c g c c c c g 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 t a t a t a a g c a g a g g g c g g g g c 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 a t a t a t t c g t c t c c

780 CMV promoter t c g t t t a g t g a a c c g t c a g a t c a c t a g t a g c t t t a t t g c g g t a g t t t a t c a c a g t t a a a t a g c a a a t c a c t t g g c a g t c t a g t g a t c a t c g a a a t a a c g c c a t c a a a t a g t g t c a a t t t a

840 chimeric intron t g c t a a c g c a g t c a g t g c t c g a c t g a t c a c a g g t a a g t a t c a a g g t t a c a a g a c a g g t t t a c g a t t g c g t c a g t c a c g a g c t g a c t a g t g t c c a t t c a t a g t t c c a a t g t t c t g t c c a a a

900 chimeric intron a a g g a g g c c a a t a g a a a c t g g g c t t g t c g a g a c a g a g a a g a t t c t t g c g t t t c t g a t a g g t t c c t c c g g t t a t c t t t g a c c c g a a c a g c t c t g t c t c t t c t a a g a a c g c a a a g a c t a t c c

960 chimeric intron c a c c t a t t g g t c t t a c t g a c a t c c a c t t t g c c t t t c t c t c c a c a g g g g t a c c g a a g c c g c g t g g a t a a c c a g a a t g a c t g t a g g t g a a a c g g a a a g a g a g g t g t c c c c a t g g c t t c g g c g

1020 T7 promoter Kozak sequence T A A T A C G A C T C A C T A T A G G G T7 *LAFQTGPGGG t a g c g c t a c c g g t c t a a t a c g a c t c a c t a t a g g g c g a a t t g g g t a c c g g g c c c g c c a c c a a t c g c g a t g g c c a g a t t a t g c t g a g t g a t a t c c c g c t t a a c c c a t g g c c c g g g c g g t g g t

1080 1 5 PAAKRVKLD (in frame with c-myc NLS) GGSGGGSG Kozak sequence c-myc NLS MASPAAKRVKLDGGSGGGSG HSAGGGLAHLQVAAAAAAAA t g g c t a g c c c c g c c g c c a a g c g c g t g a a g c t g g a c g g c g g c a g c g g c g g c g g c a g c g g c g a c c g a t c g g g g c g g c g g t t c g c g c a c t t c g a c c t g c c g c c g t c g c c g c c g c c g t c g c c g c

1140 GGSG 1 5 PAAKRVKLD (in frame with c-myc NLS) GGSGGGS c-myc NLS GGSGPAAKRVKLDGGSGGGS AAAARCSLPHFQIATAASAA g c g g c a g c g g c c c g g c a g c t a a g a g g g t g a a a c t g g a t g g c g g t a g c g g c g g a g g c a g c g c g c c g t c g c c g g g c c g t c g a t t c t c c c a c t t t g a c c t a c c g c c a t c g c c g c c t c c g t c g c

PaeR7IPspXIXhoI 1200 DKK Cas9 DKK Cas9 (in frame with c-myc NLS) GGGSGPLE 1 5 PAAKRVKLD DKK c-myc NLS Cas9 GGGSGPLEPAAKRVKLDDKK TAAAAGELRCSLPHFQIVLL g t g g c g g c a g c g g c c c c c t c g a g c c g g c a g c t a a g a g g g t g a a a c t g g a t g a c a a g a a g t c a c c g c c g t c g c c g g g g g a g c t c g g c c g t c g a t t c t c c c a c t t t g a c c t a c t g t t c t t c a

EcoRV 1260 5 10 15 20 YSIGLDIGTNSVGWAVITDE Cas9 5 10 15 20 YSIGLDIGTNSVGWAVITDE Cas9 5 10 15 20 YSIGLDIGTNSVGWAVITDE Cas9 YSIGLDIGTNSVGWAVITDE VGNPEIDTGVADAPGDNRVL a c t c c a t t g g g c t c g a t a t c g g t a c c a a c a g c g t c g g c t g g g c c g t c a t t a c g g a c g a g t t g a g g t a a c c c g a g c t a t a g c c a t g g t t g t c g c a g c c g a c c c g g c a g t a a t g c c t g c t c a

PvuI 1320 25 30 35 40 YKVPSKKFKVLGNTDRHSIK Cas9 25 30 35 40 YKVPSKKFKVLGNTDRHSIK Cas9 25 30 35 40 YKVPSKKFKVLGNTDRHSIK Cas9 YKVPSKKFKVLGNTDRHSIK VLHRAFFEFNQAIGIAVAYL a c a a g g t g c c g a g c a a a a a a t t c a a a g t t c t g g g c a a t a c c g a t c g c c a c a g c a t a a a g a t g t t c c a c g g c t c g t t t t t t a a g t t t c a a g a c c c g t t a t g g c t a g c g g t g t c g t a t t t c t

1380 45 50 55 60 KNLIGALLFDSGETAEATRL Cas9 45 50 55 60 KNLIGALLFDSGETAEATRL Cas9 45 50 55 60 KNLIGALLFDSGETAEATRL Cas9 KNLIGALLFDSGETAEATRL LVENSGEQEVGPLRGFGRPE a g a a c c t c a t t g g a g c c c t c c t g t t c g a c t c c g g g g a g a c g g c c g a a g c c a c g c g g c t c a t c t t g g a g t a a c c t c g g g a g g a c a a g c t g a g g c c c c t c t g c c g g c t t c g g t g c g c c g a g t

SbfI 1440 65 70 75 80 KRTARRRYTRRKNRICYLQE Cas9 65 70 75 80 KRTARRRYTRRKNRICYLQE Cas9 65 70 75 80 KRTARRRYTRRKNRICYLQE Cas9 KRTARRRYTRRKNRICYLQE FSCCPASIGASLIPDAVQLL a a a g a a c a g c a c g g c g c a g a t a t a c c c g c a g a a a g a a t c g g a t c t g c t a c c t g c a g g a g a t t t c t t g t c g t g c c g c g t c t a t a t g g g c g t c t t t c t t a g c c t a g a c g a t g g a c g t c c t c t

1500 85 90 95 100 IFSNEMAKVDDSFFHRLEES Cas9 85 90 95 100 IFSNEMAKVDDSFFHRLEES Cas9 85 90 95 100 IFSNEMAKVDDSFFHRLEES Cas9 IFSNEMAKVDDSFFHRLEES DKTILHSLHIVREEMPQLLG t c t t t a g t a a t g a g a t g g c t a a g g t g g a t g a c t c t t t c t t c c a t a g g c t g g a g g a g t c c t a g a a a t c a t t a c t c t a c c g a t t c c a c c t a c t g a g a a a g a a g g t a t c c g a c c t c c t c a g g a

XcmI 1560 105 110 115 120 FLVEEDKKHERHPIFGNIVD Cas9 105 110 115 120 FLVEEDKKHERHPIFGNIVD Cas9 105 110 115 120 FLVEEDKKHERHPIFGNIVD Cas9 FLVEEDKKHERHPIFGNIVD KQHLLIFLVLAVWDKAIDHV t t t t g g t g g a g g a g g a t a a a a a g c a c g a g c g c c a c c c a a t c t t t g g c a a t a t c g t g g a c g a a a a c c a c c t c c t c c t a t t t t t c g t g c t c g c g g t g g g t t a g a a a c c g t t a t a g c a c c t g c

1620 125 130 135 140 EVAYHEKYPTIYHLRKKLVD Cas9 125 130 135 140 EVAYHEKYPTIYHLRKKLVD Cas9 125 130 135 140 EVAYHEKYPTIYHLRKKLVD Cas9 EVAYHEKYPTIYHLRKKLVD LHRVMFLVWGYIM a g g t g g c g t a c c a t g a a a a g t a c c c a a c c a t a t a t c a t c t g a g g a a g a a g c t g g t a g a c a t c c a c c g c a t g g t a c t t t t c a t g g g t t g g t a t a t a g t a g a c t c c t t c t t c g a c c a t c t g t

1680 145 150 155 160 STDKADLRLIYLALAHMIKF Cas9 145 150 155 160 STDKADLRLIYLALAHMIKF Cas9 145 150 155 160 STDKADLRLIYLALAHMIKF Cas9 STDKADLRLIYLALAHMIKF g t a c t g a t a a g g c t g a c t t g c g g t t g a t c t a t c t c g c g c t g g c g c a c a t g a t c a a a t t t c c a t g a c t a t t c c g a c t g a a c g c c a a c t a g a t a g a g c g c g a c c g c g t g t a c t a g t t t a a a g

SalI 1740 165 170 175 180 RGHFLIEGDLNPDNSDVDKL Cas9 165 170 175 180 RGHFLIEGDLNPDNSDVDKL Cas9 165 170 175 180 RGHFLIEGDLNPDNSDVDKL Cas9 RGHFLIEGDLNPDNSDVDKL g g g g a c a c t t c c t c a t c g a g g g g g a c c t g a a c c c a g a c a a c a g c g a t g t c g a c a a a c t c t c c c c t g t g a a g g a g t a g c t c c c c c t g g a c t t g g g t c t g t t g t c g c t a c a g c t g t t t g a g a

1800 185 190 195 200 FIQLVQTYNQLFEENPINAS Cas9 185 190 195 200 FIQLVQTYNQLFEENPINAS Cas9 185 190 195 200 FIQLVQTYNQLFEENPINAS Cas9 FIQLVQTYNQLFEENPINAS t t a t c c a a c t g g t t c a g a c t t a c a a t c a g c t t t t c g a g g a g a a c c c g a t c a a c g c a t c c g a a t a g g t t g a c c a a g t c t g a a t g t t a g t c g a a a a g c t c c t c t t g g g c t a g t t g c g t a g g c

1860 205 210 215 220 GVDAKAILSARLSKSRRLEN Cas9 205 210 215 220 GVDAKAILSARLSKSRRLEN Cas9 205 210 215 220 GVDAKAILSARLSKSRRLEN Cas9 GVDAKAILSARLSKSRRLEN g c g t t g a c g c c a a a g c a a t c c t g a g c g c t a g g c t g t c c a a a t c c c g g c g g c t c g a a a a c c c g c a a c t g c g g t t t c g t t a g g a c t c g c g a t c c g a c a g g t t t a g g g c c g c c g a g c t t t t g g

1920 225 230 235 240 LIAQLPGEKKNGLFGNLIAL Cas9 225 230 235 240 LIAQLPGEKKNGLFGNLIAL Cas9 225 230 235 240 LIAQLPGEKKNGLFGNLIAL Cas9 LIAQLPGEKKNGLFGNLIAL t c a t c g c a c a g c t c c c t g g g g a g a a g a a g a a c g g c c t g t t t g g t a a t c t t a t c g c c c t g t a g t a g c g t g t c g a g g g a c c c c t c t t c t t c t t g c c g g a c a a a c c a t t a g a a t a g c g g g a c a

1980 245 250 255 260 SLGLTPNFKSNFDLAEDAKL Cas9 245 250 255 260 SLGLTPNFKSNFDLAEDAKL Cas9 245 250 255 260 SLGLTPNFKSNFDLAEDAKL Cas9 SLGLTPNFKSNFDLAEDAKL c a c t c g g g c t g a c c c c c a a c t t t a a a t c t a a c t t c g a c c t g g c c g a a g a t g c c a a g c t g c g t g a g c c c g a c t g g g g g t t g a a a t t t a g a t t g a a g c t g g a c c g g c t t c t a c g g t t c g a c g

2040 265 270 275 280 QLSKDTYDDDLDNLLAQIGD Cas9 265 270 275 280 QLSKDTYDDDLDNLLAQIGD Cas9 265 270 275 280 QLSKDTYDDDLDNLLAQIGD Cas9 QLSKDTYDDDLDNLLAQIGD a a c t g a g c a a a g a c a c c t a c g a t g a t g a t c t c g a c a a t c t g c t g g c c c a g a t c g g c g a c c t t g a c t c g t t t c t g t g g a t g c t a c t a c t a g a g c t g t t a g a c g a c c g g g t c t a g c c g c t g g

2100 285 290 295 300 QYADLFLAAKNLSDAILLSD Cas9 285 290 295 300 QYADLFLAAKNLSDAILLSD Cas9 285 290 295 300 QYADLFLAAKNLSDAILLSD Cas9 QYADLFLAAKNLSDAILLSD a g t a c g c a g a c c t t t t t t t g g c g g c a a a g a a c c t g t c a g a c g c c a t t c t g c t g a g t g a t a t c a t g c g t c t g g a a a a a a a c c g c c g t t t c t t g g a c a g t c t g c g g t a a g a c g a c t c a c t a t

BlpI 2160 305 310 315 320 ILRVNTEITKAPLSASMIKR Cas9 305 310 315 320 ILRVNTEITKAPLSASMIKR Cas9 305 310 315 320 ILRVNTEITKAPLSASMIKR Cas9 ILRVNTEITKAPLSASMIKR t t c t g c g a g t g a a c a c g g a g a t c a c c a a a g c t c c g c t g a g c g c t a g t a t g a t c a a g c g c t a a g a c g c t c a c t t g t g c c t c t a g t g g t t t c g a g g c g a c t c g c g a t c a t a c t a g t t c g c g a

2220 325 330 335 340 YDEHHQDLTLLKALVRQQLP Cas9 325 330 335 340 YDEHHQDLTLLKALVRQQLP Cas9 325 330 335 340 YDEHHQDLTLLKALVRQQLP Cas9 YDEHHQDLTLLKALVRQQLP a t g a t g a g c a c c a c c a a g a c t t g a c t t t g c t g a a g g c c c t t g t c a g a c a g c a a c t g c c t g t a c t a c t c g t g g t g g t t c t g a a c t g a a a c g a c t t c c g g g a a c a g t c t g t c g t t g a c g g a c

2280 345 350 355 360 EKYKEIFFDQSKNGYAGYID Cas9 345 350 355 360 EKYKEIFFDQSKNGYAGYID Cas9 345 350 355 360 EKYKEIFFDQSKNGYAGYID Cas9 EKYKEIFFDQSKNGYAGYID a g a a g t a c a a g g a a a t t t t c t t c g a t c a g t c t a a a a a t g g c t a c g c c g g a t a c a t t g a c g t c t t c a t g t t c c t t t a a a a g a a g c t a g t c a g a t t t t t a c c g a t g c g g c c t a t g t a a c t g c

2340 365 370 375 380 GGASQEEFYKFIKPILEKMD Cas9 365 370 375 380 GGASQEEFYKFIKPILEKMD Cas9 365 370 375 380 GGASQEEFYKFIKPILEKMD Cas9 GGASQEEFYKFIKPILEKMD g c g g a g c a a g c c a g g a g g a a t t t t a c a a a t t t a t t a a g c c c a t c t t g g a a a a a a t g g a c g c g c c t c g t t c g g t c c t c c t t a a a a t g t t t a a a t a a t t c g g g t a g a a c c t t t t t t a c c t g c

2400 385 390 395 400 GTEELLVKLNREDLLRKQRT Cas9 385 390 395 400 GTEELLVKLNREDLLRKQRT Cas9 385 390 395 400 GTEELLVKLNREDLLRKQRT Cas9 GTEELLVKLNREDLLRKQRT g c a c c g a g g a g c t g c t g g t a a a g c t g a a c a g a g a a g a t c t g t t g c g c a a a c a g c g c a c t t c g t g g c t c c t c g a c g a c c a t t t c g a c t t g t c t c t t c t a g a c a a c g c g t t t g t c g c g t g a a

Bsu36I 2460 405 410 415 420 FDNGSIPHQIHLGELHAILR Cas9 405 410 415 420 FDNGSIPHQIHLGELHAILR Cas9 405 410 415 420 FDNGSIPHQIHLGELHAILR Cas9 FDNGSIPHQIHLGELHAILR t c g a c a a t g g a a g c a t c c c c c a c c a g a t t c a c c t g g g c g a a c t g c a c g c t a t c c t c a g g c a g c t g t t a c c t t c g t a g g g g g t g g t c t a a g t g g a c c c g c t t g a c g t g c g a t a g g a g t c c g

2520 425 430 435 440 RQEDFYPFLKDNREKIEKIL Cas9 425 430 435 440 RQEDFYPFLKDNREKIEKIL Cas9 425 430 435 440 RQEDFYPFLKDNREKIEKIL Cas9 RQEDFYPFLKDNREKIEKIL g g c a a g a g g a t t t c t a c c c c t t t t t g a a a g a t a a c a g g g a a a a g a t t g a g a a a a t c c t c a c c g t t c t c c t a a a g a t g g g g a a a a a c t t t c t a t t g t c c c t t t t c t a a c t c t t t t a g g a g t

2580 445 450 455 460 TFRIPYYVGPLARGNSRFAW Cas9 445 450 455 460 TFRIPYYVGPLARGNSRFAW Cas9 445 450 455 460 TFRIPYYVGPLARGNSRFAW Cas9 TFRIPYYVGPLARGNSRFAW c a t t t c g g a t a c c c t a c t a t g t a g g c c c c c t c g c t c g g g g a a a t t c c a g a t t c g c g t g g a g t a a a g c c t a t g g g a t g a t a c a t c c g g g g g a g c g a g c c c c t t t a a g g t c t a a g c g c a c c t

2640 465 470 475 480 MTRKSEETITPWNFEEVVDK Cas9 465 470 475 480 MTRKSEETITPWNFEEVVDK Cas9 465 470 475 480 MTRKSEETITPWNFEEVVDK Cas9 MTRKSEETITPWNFEEVVDK *FLGDSGPVELFDHIL t g a c t c g c a a a t c a g a a g a g a c c a t c a c t c c c t g g a a c t t c g a g g a a g t c g t g g a t a a g g a c t g a g c g t t t a g t c t t c t c t g g t a g t g a g g g a c c t t g a a g c t c c t t c a g c a c c t a t t c c

2700 485 490 495 500 GASAQSFIERMTNFDKNLPN Cas9 485 490 495 500 GASAQSFIERMTNFDKNLPN Cas9 485 490 495 500 GASAQSFIERMTNFDKNLPN Cas9 GASAQSFIERMTNFDKNLPN PGRGLGEDFPHSVKIFIQRV g g g c c t c t g c c c a g t c c t t c a t c g a a a g g a t g a c t a a c t t t g a t a a a a a t c t g c c t a a c g c c c g g a g a c g g g t c a g g a a g t a g c t t t c c t a c t g a t t g a a a c t a t t t t t a g a c g g a t t g c

2760 505 510 515 520 EKVLPKHSLLYEYFTVYNEL Cas9 505 510 515 520 EKVLPKHSLLYEYFTVYNEL Cas9 505 510 515 520 EKVLPKHSLLYEYFTVYNEL Cas9 EKVLPKHSLLYEYFTVYNEL FLHKRFVRQQVLVECNIVLE a a a a g g t g c t t c c t a a a c a c t c t c t g c t g t a c g a g t a c t t c a c a g t t t a t a a c g a g c t c a t t t t c c a c g a a g g a t t t g t g a g a g a c g a c a t g c t c a t g a a g t g t c a a a t a t t g c t c g a g t

2820 525 530 535 540 TKVKYVTEGMRKPAFLSGEQ Cas9 525 530 535 540 TKVKYVTEGMRKPAFLSGEQ Cas9 525 530 535 540 TKVKYVTEGMRKPAFLSGEQ Cas9 TKVKYVTEGMRKPAFLSGEQ GLDFVDCFPHSLWCEQRSLL c c a a g g t c a a a t a c g t c a c a g a a g g g a t g a g a a a g c c a g c a t t c c t g t c t g g a g a g c a g a g g t t c c a g t t t a t g c a g t g t c t t c c c t a c t c t t t c g g t c g t a a g g a c a g a c c t c t c g t c t

2880 545 550 555 560 KKAIVDLLFKTNRKVTVKQL Cas9 545 550 555 560 KKAIVDLLFKTNRKVTVKQL Cas9 545 550 555 560 KKAIVDLLFKTNRKVTVKQL Cas9 KKAIVDLLFKTNRKVTVKQL LFSDHVEEELRVPFNGHFLE a g a a a g c t a t c g t g g a c c t c c t c t t c a a g a c g a a c c g g a a a g t t a c c g t g a a a c a g c t c a t c t t t c g a t a g c a c c t g g a g g a g a a g t t c t g c t t g g c c t t t c a a t g g c a c t t t g t c g a g t

2940 565 570 575 580 KEDYFKKIECFDSVEISGVE Cas9 565 570 575 580 KEDYFKKIECFDSVEISGVE Cas9 565 570 575 580 KEDYFKKIECFDSVEISGVE Cas9 KEDYFKKIECFDSVEISGVE FFVIEFLNFTEVRNFDASHL a a g a a g a c t a t t t c a a a a a g a t t g a a t g t t t c g a c t c t g t t g a a a t c a g c g g a g t g g a g g t t c t t c t g a t a a a g t t t t t c t a a c t t a c a a a g c t g a g a c a a c t t t a g t c g c c t c a c c t c c

3000 585 590 595 600 DRFNASLGTYHDLLKIIKDK Cas9 585 590 595 600 DRFNASLGTYHDLLKIIKDK Cas9 585 590 595 600 DRFNASLGTYHDLLKIIKDK Cas9 DRFNASLGTYHDLLKIIKDK IAEVCGQSRIVIEQFDNFVL a t c g c t t c a a c g c a t c c c t g g g a a c g t a t c a c g a t c t c c t g a a a a t c a t t a a a g a c a a g g t a g c g a a g t t g c g t a g g g a c c c t t g c a t a g t g c t a g a g g a c t t t t a g t a a t t t c t g t t c c

3060 605 610 615 620 DFLDNEENEDILEDIVLTLT Cas9 605 610 615 620 DFLDNEENEDILEDIVLTLT Cas9 605 610 615 620 DFLDNEENEDILEDIVLTLT Cas9 DFLDNEENEDILEDIVLTLT VEQVILLVLVNKLVNDEGKR a c t t c c t g g a c a a t g a g g a g a a c g a g g a c a t t c t t g a g g a c a t t g t c c t c a c c c t t a c g t t g a a g g a c c t g t t a c t c c t c t t g c t c c t g t a a g a a c t c c t g t a a c a g g a g t g g g a a t g c a

3120 625 630 635 640 LFEDREMIEERLKTYAHLFD Cas9 625 630 635 640 LFEDREMIEERLKTYAHLFD Cas9 625 630 635 640 LFEDREMIEERLKTYAHLFD Cas9 LFEDREMIEERLKTYAHLFD QKFIPLHNFFAQFSVSM t g t t t g a a g a t a g g g a g a t g a t t g a a g a a c g c t t g a a a a c t t a c g c t c a t c t c t t c g a c g a c a a a c t t c t a t c c c t c t a c t a a c t t c t t g c g a a c t t t t g a a t g c g a g t a g a g a a g c t g c

BspHI 3180 645 650 655 660 DKVMKQLKRRRYTGWGRLSR Cas9 645 650 655 660 DKVMKQLKRRRYTGWGRLSR Cas9 645 650 655 660 DKVMKQLKRRRYTGWGRLSR Cas9 DKVMKQLKRRRYTGWGRLSR *S a c a a a g t c a t g a a a c a g c t c a a g a g a c g c c g a t a t a c a g g a t g g g g g c g g c t g t c a a g a a t g t t t c a g t a c t t t g t c g a g t t c t c t g c g g c t a t a t g t c c t a c c c c c g c c g a c a g t t c t t

3240 665 670 675 680 KLINGIRDKQSGKTILDFLK Cas9 665 670 675 680 KLINGIRDKQSGKTILDFLK Cas9 665 670 675 680 KLINGIRDKQSGKTILDFLK Cas9 KLINGIRDKQSGKTILDFLK FQDIADSVLLTSLCDQIKKL a a c t g a t c a a t g g c a t c c g a g a c a a g c a g a g t g g a a a g a c a a t c c t g g a t t t t c t t a a g t t t g a c t a g t t a c c g t a g g c t c t g t t c g t c t c a c c t t t c t g t t a g g a c c t a a a a g a a t t c a

3300 685 690 695 700 SDGFANRNFMQLIHDDSLTF Cas9 685 690 695 700 SDGFANRNFMQLIHDDSLTF Cas9 685 690 695 700 SDGFANRNFMQLIHDDSLTF Cas9 SDGFANRNFMQLIHDDSLTF GISKGVPVEHLQDMIVREGK c c g a t g g a t t t g c c a a c c g g a a c t t c a t g c a g t t g a t c c a t g a t g a c t c t c t c a c c t t t a g g c t a c c t a a a c g g t t g g c c t t g a a g t a c g t c a a c t a g g t a c t a c t g a g a g a g t g g a a a t

3360 705 710 715 720 KEDIQKAQVSGQGDSLHEHI Cas9 705 710 715 720 KEDIQKAQVSGQGDSLHEHI Cas9 705 710 715 720 KEDIQKAQVSGQGDSLHEHI Cas9 KEDIQKAQVSGQGDSLHEHI LLVDLFCLNRALPVTKVLVD a g g a g g a c a t c c a g a a a g c a c a a g t t t c t g g c c a g g g g g a c a g t c t t c a c g a g c a c a t c g t c c t c c t g t a g g t c t t t c g t g t t c a a a g a c c g g t c c c c c t g t c a g a a g t g c t c g t g t a g c

3420 725 730 735 740 ANLAGSPAIKKGILQTVKVV Cas9 725 730 735 740 ANLAGSPAIKKGILQTVKVV Cas9 725 730 735 740 ANLAGSPAIKKGILQTVKVV Cas9 ANLAGSPAIKKGILQTVKVV SIKCTAWSDFLSYQLGNLDH c t a a t c t t g c a g g t a g c c c a g c t a t c a a a a a g g g a a t a c t g c a g a c c g t t a a g g t c g t g g g a t t a g a a c g t c c a t c g g g t c g a t a g t t t t t c c c t t a t g a c g t c t g g c a a t t c c a g c a c c

3480 745 750 755 760 DELVKVMGRHKPENIVIEMA Cas9 745 750 755 760 DELVKVMGRHKPENIVIEMA Cas9 745 750 755 760 DELVKVMGRHKPENIVIEMA Cas9 DELVKVMGRHKPENIVIEMA IFEDFYHSPMLGLIDNDLHG a t g a a c t c g t c a a a g t a a t g g g a a g g c a t a a g c c c g a g a a t a t c g t t a t c g a g a t g g c c c t a c t t g a g c a g t t t c a t t a c c c t t c c g t a t t c g g g c t c t t a t a g c a a t a g c t c t a c c g g g

3540 765 770 775 780 RENQTTQKGQKNSRERMKRI Cas9 765 770 775 780 RENQTTQKGQKNSRERMKRI Cas9 765 770 775 780 RENQTTQKGQKNSRERMKRI Cas9 RENQTTQKGQKNSRERMKRI SLVLSGLLSLLVTPFPHLPN g a g a g a a c c a a a c t a c c c a g a a g g g a c a g a a g a a c a g t a g g g a a a g g a t g a a g a g g a t t g c t c t c t t g g t t t g a t g g g t c t t c c c t g t c t t c t t g t c a t c c c t t t c c t a c t t c t c c t a a c

3600 785 790 795 800 EEGIKELGSQILKEHPVENT Cas9 785 790 795 800 EEGIKELGSQILKEHPVENT Cas9 785 790 795 800 EEGIKELGSQILKEHPVENT Cas9 EEGIKELGSQILKEHPVENT FLTYFFQPGLDKLFVWNFVG a a g a g g g t a t a a a a g a a c t g g g g t c c c a a a t c c t t a a g g a a c a c c c a g t t g a a a a c a c c c t t c t c c c a t a t t t t c t t g a c c c c a g g g t t t a g g a a t t c c t t g t g g g t c a a c t t t t g t g g g

3660 805 810 815 820 QLQNEKLYLYYLQNGRDMYV Cas9 805 810 815 820 QLQNEKLYLYYLQNGRDMYV Cas9 805 810 815 820 QLQNEKLYLYYLQNGRDMYV Cas9 QLQNEKLYLYYLQNGRDMYV LKLILLEVQVVQLVAPVHVH a g c t t c a g a a t g a g a a g c t c t a c c t g t a c t a c c t g c a g a a c g g c a g g g a c a t g t a c g t g g t c g a a g t c t t a c t c t t c g a g a t g g a c a t g a t g g a c g t c t t g c c g t c c c t g t a c a t g c a c c

HindIIIBmgBI 3720 825 830 835 840 DQELDINRLSDYDVDHIVPQ Cas9 825 830 835 840 DQELDINRLSDYDVDHIVPQ Cas9 825 830 835 840 DQELDINRLSDYDVDHIVPQ Cas9 DQELDINRLSDYDVDHIVPQ ILFQVDVPQGVVVHIM a t c a g g a a c t g g a c a t c a a c c g g t t g t c c g a c t a c g a c g t g g a t c a t a t c g t g c c c c a a a t a g t c c t t g a c c t g t a g t t g g c c a a c a g g c t g a t g c t g c a c c t a g t a t a g c a c g g g g t t t

3780 845 850 855 860 SFLKDDSIDNKVLTRSDKNR Cas9 845 850 855 860 SFLKDDSIDNKVLTRSDKNR Cas9 845 850 855 860 SFLKDDSIDNKVLTRSDKNR Cas9 SFLKDDSIDNKVLTRSDKNR g c t t t c t c a a a g a t g a t t c t a t t g a t a a t a a a g t g t t g a c a a g a t c c g a t a a a a a t a g a g c g a a a g a g t t t c t a c t a a g a t a a c t a t t a t t t c a c a a c t g t t c t a g g c t a t t t t t a t c t c

3840 865 870 875 880 GKSDNVPSEEVVKKMKNYWR Cas9 865 870 875 880 GKSDNVPSEEVVKKMKNYWR Cas9 865 870 875 880 GKSDNVPSEEVVKKMKNYWR Cas9 GKSDNVPSEEVVKKMKNYWR g g a a g a g t g a t a a c g t c c c c t c a g a a g a a g t t g t c a a g a a a a t g a a a a a t t a t t g g c g g c c c t t c t c a c t a t t g c a g g g g a g t c t t c t t c a a c a g t t c t t t t a c t t t t t a a t a a c c g c c g

3900 885 890 895 900 QLLNAKLITQRKFDNLTKAE Cas9 885 890 895 900 QLLNAKLITQRKFDNLTKAE Cas9 885 890 895 900 QLLNAKLITQRKFDNLTKAE Cas9 QLLNAKLITQRKFDNLTKAE a g c t g c t g a a c g c c a a a c t g a t c a c a c a a c g g a a g t t c g a t a a t c t g a c t a a g g c t g a a c t c g a c g a c t t g c g g t t t g a c t a g t g t g t t g c c t t c a a g c t a t t a g a c t g a t t c c g a c t t g

3960 905 910 915 920 RGGLSELDKAGFIKRQLVET Cas9 905 910 915 920 RGGLSELDKAGFIKRQLVET Cas9 905 910 915 920 RGGLSELDKAGFIKRQLVET Cas9 RGGLSELDKAGFIKRQLVET g a g g t g g c c t g t c t g a g t t g g a t a a a g c c g g c t t c a t c a a a a g g c a g c t t g t t g a g a c a c c t c c a c c g g a c a g a c t c a a c c t a t t t c g g c c g a a g t a g t t t t c c g t c g a a c a a c t c t g t g

4020 925 930 935 940 RQITKHVAQILDSRMNTKYD Cas9 925 930 935 940 RQITKHVAQILDSRMNTKYD Cas9 925 930 935 940 RQITKHVAQILDSRMNTKYD Cas9 RQITKHVAQILDSRMNTKYD g c c a g a t c a c c a a g c a c g t g g c c c a a a t t c t c g a t t c a c g c a t g a a c a c c a a g t a c g a t g c g g t c t a g t g g t t c g t g c a c c g g g t t t a a g a g c t a a g t g c g t a c t t g t g g t t c a t g c t a c

4080 945 950 955 960 ENDKLIREVKVITLKSKLVS Cas9 945 950 955 960 ENDKLIREVKVITLKSKLVS Cas9 945 950 955 960 ENDKLIREVKVITLKSKLVS Cas9 ENDKLIREVKVITLKSKLVS * a a a a t g a c a a a c t g a t t c g a g a g g t g a a a g t t a t t a c t c t g a a g t c t a a g c t g g t c t c a g t t t t a c t g t t t g a c t a a g c t c t c c a c t t t c a a t a a t g a g a c t t c a g a t t c g a c c a g a g t c

FspAI 4140 965 970 975 980 DFRKDFQFYKVREINNYHHA Cas9 965 970 975 980 DFRKDFQFYKVREINNYHHA Cas9 965 970 975 980 DFRKDFQFYKVREINNYHHA Cas9 DFRKDFQFYKVREINNYHHA IESLVKLKILHSLDVIVVMR a t t t c a g a a a g g a c t t t c a g t t t t a t a a g g t g a g a g a g a t c a a c a a t t a c c a c c a t g c g c t a a a g t c t t t c c t g a a a g t c a a a a t a t t c c a c t c t c t c t a g t t g t t a a t g g t g g t a c g c g

4200 985 990 995 1000 HDAYLNAVVGTALIKKYPKL Cas9 985 990 995 1000 HDAYLNAVVGTALIKKYPKL Cas9 985 990 995 1000 HDAYLNAVVGTALIKKYPKL Cas9 HDAYLNAVVGTALIKKYPKL MIGVQICHYASCKDFFIGLQ a t g a t g c c t a c c t g a a t g c a g t g g t a g g c a c t g c a c t t a t c a a a a a a t a t c c c a a g c t g g t a c t a c g g a t g g a c t t a c g t c a c c a t c c g t g a c g t g a a t a g t t t t t t a t a g g g t t c g a c c

4260 1005 1010 1015 1020 ESEFVYGDYKVYDVRKMIAK Cas9 1005 1010 1015 1020 ESEFVYGDYKVYDVRKMIAK Cas9 1005 1010 1015 1020 ESEFVYGDYKVYDVRKMIAK Cas9 ESEFVYGDYKVYDVRKMIAK FRFKNVSVIFHVINPFHDCL a a t c t g a a t t t g t t t a c g g a g a c t a t a a a g t g t a c g a t g t t a g g a a a a t g a t c g c a a a g t t t a g a c t t a a a c a a a t g c c t c t g a t a t t t c a c a t g c t a c a a t c c t t t t a c t a g c g t t t c a

XmnI 4320 1025 1030 1035 1040 SEQEIGKATAKYFFYSNIMN Cas9 1025 1030 1035 1040 SEQEIGKATAKYFFYSNIMN Cas9 1025 1030 1035 1040 SEQEIGKATAKYFFYSNIMN Cas9 SEQEIGKATAKYFFYSNIMN RLLFYALGGSLVEKVAINHI c t g a g c a g g a a a t a g g c a a g g c c a c c g c t a a g t a c t t c t t t t a c a g c a a t a t t a t g a a t t g a c t c g t c c t t t a t c c g t t c c g g t g g c g a t t c a t g a a g a a a a t g t c g t t a t a a t a c t t a a

4380 1045 1050 1055 1060 FFKTEITLANGEIRKRPLIE Cas9 1045 1050 1055 1060 FFKTEITLANGEIRKRPLIE Cas9 1045 1050 1055 1060 FFKTEITLANGEIRKRPLIE Cas9 FFKTEITLANGEIRKRPLIE KELGLNCQGISLNPLSWKDF t t t t c a a g a c c g a g a t t a c a c t g g c c a a t g g a g a g a t t c g g a a g c g a c c a c t t a t c g a a a a a a a g t t c t g g c t c t a a t g t g a c c g g t t a c c t c t c t a a g c c t t c g c t g g t g a a t a g c t t t

NruI 4440 1065 1070 1075 1080 TNGETGEIVWDKGRDFATVR Cas9 1065 1070 1075 1080 TNGETGEIVWDKGRDFATVR Cas9 1065 1070 1075 1080 TNGETGEIVWDKGRDFATVR Cas9 TNGETGEIVWDKGRDFATVR CVSFCSFDHPVLTPIERCDA c a a a c g g a g a a a c a g g a g a a a t c g t g t g g g a c a a g g g t a g g g a t t t c g c g a c a g t c c g c a g t t t g c c t c t t t g t c c t c t t t a g c a c a c c c t g t t c c c a t c c c t a a a g c g c t g t c a g g c g t

PaqCI 4500 1085 1090 1095 1100 KVLSMPQVNIVKKTEVQTGG Cas9 1085 1090 1095 1100 KVLSMPQVNIVKKTEVQTGG Cas9 1085 1090 1095 1100 KVLSMPQVNIVKKTEVQTGG Cas9 KVLSMPQVNIVKKTEVQTGG LDQGHRLHVDNFLGFYLGSA a g g t c c t g t c c a t g c c g c a g g t g a a c a t c g t t a a a a a g a c c g a a g t a c a g a c c g g a g g c t t c c a g g a c a g g t a c g g c g t c c a c t t g t a g c a a t t t t t c t g g c t t c a t g t c t g g c c t c c g a

4560 1105 1110 1115 1120 FSKESILPKRNSDKLIARKK Cas9 1105 1110 1115 1120 FSKESILPKRNSDKLIARKK Cas9 1105 1110 1115 1120 FSKESILPKRNSDKLIARKK Cas9 FSKESILPKRNSDKLIARKK EGLFTDERFPVAVLQDCAFF t c t c c a a g g a a a g t a t c c t c c c g a a a a g g a a c a g c g a c a a g c t g a t c g c a c g c a a a a a a g a g a g g t t c c t t t c a t a g g a g g g c t t t t c c t t g t c g c t g t t c g a c t a g c g t g c g t t t t t t c

4620 1125 1130 1135 1140 DWDPKKYGGFDSPTVAYSVL Cas9 1125 1130 1135 1140 DWDPKKYGGFDSPTVAYSVL Cas9 1125 1130 1135 1140 DWDPKKYGGFDSPTVAYSVL Cas9 DWDPKKYGGFDSPTVAYSVL IPVGLFVASEIRRCDSVTYQ a t t g g g a c c c c a a g a a a t a c g g c g g a t t c g a t t c t c c t a c a g t c g c t t a c a g t g t a c t g g t a a c c c t g g g g t t c t t t a t g c c g c c t a a g c t a a g a g g a t g t c a g c g a a t g t c a c a t g a c c

4680 1145 1150 1155 1160 VVAKVEKGKSKKLKSVKELL Cas9 1145 1150 1155 1160 VVAKVEKGKSKKLKSVKELL Cas9 1145 1150 1155 1160 VVAKVEKGKSKKLKSVKELL Cas9 VVAKVEKGKSKKLKSVKELL NHGFHLFPLRFFEFADLFQQ t t g t g g c c a a a g t g g a g a a a g g g a a g t c t a a a a a a c t c a a a a g c g t c a a g g a a c t g c t g g a a c a c c g g t t t c a c c t c t t t c c c t t c a g a t t t t t t g a g t t t t c g c a g t t c c t t g a c g a c c

4740 1165 1170 1175 1180 GITIMERSSFEKNPIDFLEA Cas9 1165 1170 1175 1180 GITIMERSSFEKNPIDFLEA Cas9 1165 1170 1175 1180 GITIMERSSFEKNPIDFLEA Cas9 GITIMERSSFEKNPIDFLEA ADCDHLSGAELFVGDVKEFR g c a t c a c a a t c a t g g a g c g a t c c a g c t t c g a g a a a a a c c c c a t c g a c t t t c t c g a a g c g a c g t a g t g t t a g t a c c t c g c t a g g t c g a a g c t c t t t t t g g g g t a g c t g a a a g a g c t t c g c t

4800 1185 1190 1195 1200 KGYKEVKKDLIIKLPKYSLF Cas9 1185 1190 1195 1200 KGYKEVKKDLIIKLPKYSLF Cas9 1185 1190 1195 1200 KGYKEVKKDLIIKLPKYSLF Cas9 KGYKEVKKDLIIKLPKYSLF FSIFLDFFVEDNLQGLVREK a a g g a t a t a a a g a g g t c a a a a a a g a c c t c a t c a t t a a g c t g c c c a a g t a c t c t c t c t t t g t t c c t a t a t t t c t c c a g t t t t t t c t g g a g t a g t a a t t c g a c g g g t t c a t g a g a g a g a a a c

4860 1205 1210 1215 1220 ELENGRKRMLASAGELQKGN Cas9 1205 1210 1215 1220 ELENGRKRMLASAGELQKGN Cas9 1205 1210 1215 1220 ELENGRKRMLASAGELQKGN Cas9 ELENGRKRMLASAGELQKGN LKFVAPFSHESTRALQLFTV a g c t t g a a a a c g g c c g g a a a c g a a t g c t c g c t a g t g c g g g c g a g c t g c a g a a a g g t a a c g t c g a a c t t t t g c c g g c c t t t g c t t a c g a g c g a t c a c g c c c g c t c g a c g t c t t t c c a t t g c

4920 1225 1230 1235 1240 ELALPSKYVNFLYLASHYEK Cas9 1225 1230 1235 1240 ELALPSKYVNFLYLASHYEK Cas9 1225 1230 1235 1240 ELALPSKYVNFLYLASHYEK Cas9 ELALPSKYVNFLYLASHYEK LQCQGRFVNIEQIQGAVIFL a g c t g g c a c t g c c c t c t a a a t a c g t t a a t t t c t t g t a t c t g g c c a g c c a c t a t g a a a a g c t c g a c c g t g a c g g g a g a t t t a t g c a a t t a a a g a a c a t a g a c c g g t c g g t g a t a c t t t t c g

4980 1245 1250 1255 1260 LKGSPEDNEQKQLFVEQHKH Cas9 1245 1250 1255 1260 LKGSPEDNEQKQLFVEQHKH Cas9 1245 1250 1255 1260 LKGSPEDNEQKQLFVEQHKH Cas9 LKGSPEDNEQKQLFVEQHKH EFPRGFIILLLLQEHFLVFV t c a a a g g g t c t c c c g a a g a t a a t g a g c a g a a g c a g c t g t t c g t g g a a c a a c a c a a a c a c t a g t t t c c c a g a g g g c t t c t a t t a c t c g t c t t c g t c g a c a a g c a c c t t g t t g t g t t t g t g a

5040 1265 1270 1275 1280 YLDEIIEQISEFSKRVILAD Cas9 1265 1270 1275 1280 YLDEIIEQISEFSKRVILAD Cas9 1265 1270 1275 1280 YLDEIIEQISEFSKRVILAD Cas9 YLDEIIEQISEFSKRVILAD VKILDDLLYALEGFSHDEGV a c c t t g a t g a g a t c a t c g a g c a a a t a a g c g a g t t c t c c a a a a g a g t g a t c c t c g c c g a c g t g g a a c t a c t c t a g t a g c t c g t t t a t t c g c t c a a g a g g t t t t c t c a c t a g g a g c g g c t g c

5100 1285 1290 1295 1300 ANLDKVLSAYNKHRDKPIRE Cas9 1285 1290 1295 1300 ANLDKVLSAYNKHRDKPIRE Cas9 1285 1290 1295 1300 ANLDKVLSAYNKHRDKPIRE Cas9 ANLDKVLSAYNKHRDKPIRE SVEILHKRSVILVPILGDPL c t a a c c t c g a t a a g g t g c t t t c t g c t t a c a a t a a g c a c a g g g a t a a g c c c a t c a g g g a g c g a t t g g a g c t a t t c c a c g a a a g a c g a a t g t t a t t c g t g t c c c t a t t c g g g t a g t c c c t c g

BssHIIAscI 5160 1305 1310 1315 1320 QAENIIHLFTLTNLGAPAAF Cas9 1305 1310 1315 1320 QAENIIHLFTLTNLGAPAAF Cas9 1305 1310 1315 1320 QAENIIHLFTLTNLGAPAAF Cas9 QAENIIHLFTLTNLGAPAAF LCFVNDVQKSQGVQARRCGE a g g c a g a a a a c a t t a t c c a c t t g t t t a c t c t g a c c a a c t t g g g c g c g c c t g c a g c c t t c a t c c g t c t t t t g t a a t a g g t g a a c a a a t g a g a c t g g t t g a a c c c g c g c g g a c g t c g g a a g t

5220 1325 1330 1335 1340 KYFDTTIDRKRYTSTKEVLD Cas9 1325 1330 1335 1340 KYFDTTIDRKRYTSTKEVLD Cas9 1325 1330 1335 1340 KYFDTTIDRKRYTSTKEVLD Cas9 KYFDTTIDRKRYTSTKEVLD LVEVGGYVSLPVGRCLLDQV a g t a c t t c g a c a c c a c c a t a g a c a g a a a g c g g t a c a c c t c t a c a a a g g a g g t c c t g g a c g t c a t g a a g c t g t g g t g g t a t c t g t c t t t c g c c a t g t g g a g a t g t t t c c t c c a g g a c c t g c

5280 1345 1350 1355 1360 ATLIHQSITGLYETRIDLSQ Cas9 1345 1350 1355 1360 ATLIHQSITGLYETRIDLSQ Cas9 1345 1350 1355 1360 ATLIHQSITGLYETRIDLSQ Cas9 ATLIHQSITGLYETRIDLSQ GCQNM c c a c a c t g a t t c a t c a g t c a a t t a c g g g g c t c t a t g a a a c a a g a a t c g a c c t c t c t c a g c g g t g t g a c t a a g t a g t c a g t t a a t g c c c c g a g a t a c t t t g t t c t t a g c t g g a g a g a g t c g

BstZ17ISgrAI 5340 1365 LGGD (in frame with Cas9) GTGGPKKKRKVYPYDV Cas9 1365 LGGD (in frame with Cas9) GTGGPKKKRKVYPYDV Cas9 1365 LGGD GTGG 130 PKKKRKV 1 5 YPYDV Cas9 SV40 NLS 3xHA LGGDGTGGPKKKRKVYPYDV t c g g t g g a g a c g g c a c c g g c g g g c c c a a g a a g a a g a g g a a g g t a t a c c c a t a c g a t g t t c a g c c a c c t c t g c c g t g g c c g c c c g g g t t c t t c t t c t c c t t c c a t a t g g g t a t g c t a c a a g

5400 (in frame with Cas9) PDYAGYPYDVPDYAGSYPYD (in frame with Cas9) PDYAGYPYDVPDYAGSYPYD 10 15 20 25 PDYAGYPYDVPDYAGSYPYD 3xHA PDYAGYPYDVPDYAGSYPYD c t g a c t a t g c g g g c t a t c c c t a t g a c g t c c c g g a c t a t g c a g g a t c g t a t c c t t a t g a c g g a c t g a t a c g c c c g a t a g g g a t a c t g c a g g g c c t g a t a c g t c c t a g c a t a g g a a t a c t g c

BstBI 5460 (in frame with Cas9) VPDYAGSAAPAAKKKKLDFE (in frame with Cas9) VPDYAGSAAPAAKKKKLDFE 30 VPDYA GSAA 1 5 PAAKKKKLD FE 3xHA NLS VPDYAGSAAPAAKKKKLDFE t t c c a g a t t a c g c t g g a t c c g c c g c t c c g g c a g c t a a g a a a a a g a a a c t g g a t t t c g a a t a a g g t c t a a t g c g a c c t a g g c g g c g a g g c c g t c g a t t c t t t t t c t t t g a c c t a a a g c t t a

SfiIEcoRI 5520 (in frame with Cas9) SGEFGAQPARSMAEAGITGT (in frame with Cas9) SGEFGAQPARSMAEAGITGT (in frame with NLS) SGEFGAQPARSM 1 5 AEAGITGT mSA SGEFGAQPARSMAEAGITGT c c g g a g a a t t c g g g g c c c a g c c g g c c a g a t c t a t g g c g g a a g c g g g t a t c a c c g g c a c g t g g c c t c t t a a g c c c c g g g t c g g c c g g t c t a g a t a c c g c c t t c g c c c a t a g t g g c c g t g c a

BstEII 5580 (in frame with Cas9) WYNQSGSTFTVTAGADGNLT (in frame with Cas9) WYNQSGSTFTVTAGADGNLT 10 15 20 25 WYNQSGSTFTVTAGADGNLT mSA WYNQSGSTFTVTAGADGNLT g g t a c a a c c a g t c t g g t t c t a c c t t c a c c g t t a c c g c g g g t g c g g a c g g t a a c c t g a c c g c c a t g t t g g t c a g a c c a a g a t g g a a g t g g c a a t g g c g c c c a c g c c t g c c a t t g g a c t g g c

5640 (in frame with Cas9) GQYENRAQGTGCQNSPYTLT (in frame with Cas9) GQYENRAQGTGCQNSPYTLT 30 35 40 45 GQYENRAQGTGCQNSPYTLT mSA GQYENRAQGTGCQNSPYTLT g t c a g t a c g a a a a c c g t g c g c a g g g c a c t g g t t g c c a g a a c t c t c c g t a c a c c c t g a c c g c a g t c a t g c t t t t g g c a c g c g t c c c g t g a c c a a c g g t c t t g a g a g g c a t g t g g g a c t g g c

5700 (in frame with Cas9) GRYNGTKLEWRVEWNNSTEN (in frame with Cas9) GRYNGTKLEWRVEWNNSTEN 50 55 60 65 GRYNGTKLEWRVEWNNSTEN mSA GRYNGTKLEWRVEWNNSTEN g t c g t t a c a a c g g t a c c a a a c t g g a a t g g c g t g t t g a a t g g a a c a a c t c t a c c g a a a a c t c a g c a a t g t t g c c a t g g t t t g a c c t t a c c g c a c a a c t t a c c t t g t t g a g a t g g c t t t t g a

5760 (in frame with Cas9) CHSRTEWRGQYQGGAEARIN (in frame with Cas9) CHSRTEWRGQYQGGAEARIN 70 75 80 85 CHSRTEWRGQYQGGAEARIN mSA CHSRTEWRGQYQGGAEARIN g c c a c t c t c g t a c c g a a t g g c g t g g t c a g t a c c a g g g t g g t g c g g a a g c g c g t a t c a a c a c g g t g a g a g c a t g g c t t a c c g c a c c a g t c a t g g t c c c a c c a c g c c t t c g c g c a t a g t t g t

DraIII 5820 (in frame with Cas9) TQWNLTYEGGSGPATEQGQD (in frame with Cas9) TQWNLTYEGGSGPATEQGQD 90 95 100 105 TQWNLTYEGGSGPATEQGQD mSA TQWNLTYEGGSGPATEQGQD c c c a g t g g a a c c t g a c c t a c g a a g g t g g t t c t g g t c c g g c g a c c g a a c a g g g t c a g g a c a g g g t c a c c t t g g a c t g g a t g c t t c c a c c a a g a c c a g g c c g c t g g c t t g t c c c a g t c c t g t

XbaITspMIXmaISmaI 5880 (in frame with Cas9) TFTKVKPSAASGSPGGSTSS (in frame with Cas9) TFTKVKPSAASGSPGGSTSS 110 TFTKVK (in frame with mSA) PSAASGSPGGSTSS mSA SK primer C T A G G T G A T C A A G A T pBluescriptSK TFTKVKPSAASGSPGGSTSS c c t t c a c c a a a g t t a a a c c g t c t g c g g c g t c t g g a t c c c c c g g g g g a t c c a c t a g t t c t a g g a a g t g g t t t c a a t t t g g c a g a c g c c g c a g a c c t a g g g g g c c c c c t a g g t g a t c a a g a t

5940 (in frame with Cas9) RGSAAPAAKRVKLDGGSGGG (in frame with Cas9) RGSAAPAAKRVKLDGGSGGG (in frame with mSA) RGSAA 1 5 PAAKRVKLD (in frame with c-myc NLS) GGSGGG c-myc NLS SK primer C T pBluescriptSK RGSAAPAAKRVKLDGGSGGG g a g g a t c c g c c g c t c c c g c c g c c a a g c g c g t g a a g c t g g a c g g c g g c a g c g g c g g c g g c a c t c c t a g g c g g c g a g g g c g g c g g t t c g c g c a c t t c g a c c t g c c g c c g t c g c c g c c g c c g t

NotI 6000 (in frame with Cas9) SGGGSGSGPAAKRVKLD* (in frame with Cas9) SGGGSGSGPAAKRVKLD* (in frame with c-myc NLS) SGGGSGSG 1 5 PAAKRVKLD * c-myc NLS SGGGSGSGPAAKRVKLD* g c g g c g g c g g c a g c g g c t c c g g a c c g g c a g c t a a g a g g g t g a a a c t g g a t t g a t a a g c g g c g c c g c c g c c g t c g c c g a g g c c t g g c c g t c g a t t c t c c c a c t t t g a c c t a a c t a t t c g c c

AleI 6060 C T A C T C A EBV-rev c c g c c a c c g c g g t g g a g c t c c a g c t t t t g t t c c c t t t a g t g a g g g t t a a t c t c g a t g a g t g g c g g t g g c g c c a c c t c g a g g t c g a a a a c a a g g g a a a t c a c t c c c a a t t a g a g c t a c t c a

6120 SV40 poly(A) signal G A A A T T T G T G A T G SV40pA-R A A C C T G T T T G G T G EBV-rev t t g g a c a a a c c a c a a c t a g a a t g c a g t g a a a a a a a t g c t t t a t t t g t g a a a t t t g t g a t g a a c c t g t t t g g t g t t g a t c t t a c g t c a c t t t t t t t a c g a a a t a a a c a c t t t a a a c a c t a c

MfeIHpaI 6180 SV40 poly(A) signal C T A T T G C SV40pA-R c t a t t g c t t t a t t t g t a a c c a t t a t a a g c t g c a a t a a a c a a g t t a a c a a c a a c a a t t g c a g a t a a c g a a a t a a a c a t t g g t a a t a t t c g a c g t t a t t t g t t c a a t t g t t g t t g t t a a c g t

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

StuI 6300 t c t a c a a a t g t g g t a c t t a a g a g g g g g a g a c c a a a g g g c g a g a c g t t a a g g c c t c a c g t g a g a t g t t t a c a c c a t g a a t t c t c c c c c t c t g g t t t c c c g c t c t g c a a t t c c g g a g t g c a c

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

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

6480 ori T T T C T A T G G T C C G C A A A G G G pBR322ori-F g g c g a a a c c c g a c a g g a c t a t a a a g a t a c c a g g c g t t t c c c c c t g g a a g c t c c c t c g t g c c c g c t t t g g g c t g t c c t g a t a t t t c t a t g g t c c g c a a a g g g g g a c c t t c g a g g g a g c a c g

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

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

ApaLI 6660 ori c c a a g c t g g g c t g t g t g c a c g a a c c c c c c g t t c a g c c c g a c c g c t g c g c c t t a t c c g g t a g g t t c g a c c c g a c a c a c g t g c t t g g g g g g c a a g t c g g g c t g g c g a c g c g g a a t a g g c c a t

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

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

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

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

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

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

7080 265 260 255 250 *FFEDLLRYFAIRQSDP NeoR/KanR 265 260 255 250 *FFEDLLRYFAIRQSDP NeoR/KanR *FFEDLLRYFAIRQSDP t c a t g c c g t c t c a g a a g a a c t c g t c a a g a a g g c g a t a g a a g g c g a t g c g c t g c g a a t c g g a g t a c g g c a g a g t c t t c t t g a g c a g t t c t t c c g c t a t c t t c c g c t a c g c g a c g c t t a g c c

7140 245 240 235 230 AAIGYLVLFRDAWEGGLEEA NeoR/KanR 245 240 235 230 AAIGYLVLFRDAWEGGLEEA NeoR/KanR AAIGYLVLFRDAWEGGLEEA g a g c g g c g a t a c c g t a a a g c a c g a g g a a g c g g t c a g c c c a t t c g c c g c c a a g c t c t t c a g c t c g c c g c t a t g g c a t t t c g t g c t c c t t c g c c a g t c g g g t a a g c g g c g g t t c g a g a a g t c

7200 225 220 215 210 IDRTALAIDQYRDAVGLRGC NeoR/KanR 225 220 215 210 IDRTALAIDQYRDAVGLRGC NeoR/KanR T T A T A G T G C C C A T C G G T T G C Neo-F IDRTALAIDQYRDAVGLRGC c a a t a t c a c g g g t a g c c a a c g c t a t g t c c t g a t a g c g g t c c g c c a c a c c c a g c c g g c c a c g t t a t a g t g c c c a t c g g t t g c g a t a c a g g a c t a t c g c c a g g c g g t g t g g g t c g g c c g g t g

7260 205 200 195 190 DIFGSFRGNEVMINPLCADG NeoR/KanR 205 200 195 190 DIFGSFRGNEVMINPLCADG NeoR/KanR DIFGSFRGNEVMINPLCADG a g t c g a t g a a t c c a g a a a a g c g g c c a t t t t c c a c c a t g a t a t t c g g c a a g c a g g c a t c g c t c a g c t a c t t a g g t c t t t t c g c c g g t a a a a g g t g g t a c t a t a a g c c g t t c g t c c g t a g c g

SphI 7320 185 180 175 170 HTVVLDEGDPMSAKLRAFLE NeoR/KanR 185 180 175 170 HTVVLDEGDPMSAKLRAFLE NeoR/KanR HTVVLDEGDPMSAKLRAFLE c a t g g g t c a c g a c g a g a t c c t c g c c g t c g g g c a t g c t c g c c t t g a g c c t g g c g a a c a g t t g t a c c c a g t g c t g c t c t a g g a g c g g c a g c c c g t a c g a g c g g a a c t c g g a c c g c t t g t c a a

7380 165 160 155 150 APALGQHEEDLDDQDVLGAE NeoR/KanR 165 160 155 150 APALGQHEEDLDDQDVLGAE NeoR/KanR APALGQHEEDLDDQDVLGAE c g g c t g g c g c g a g c c c c t g a t g c t c t t c g t c c a g a t c a t c c t g a t c g a c a a g a c c g g c t t g c c g a c c g c g c t c g g g g a c t a c g a g a a g c a g g t c t a g t a g g a c t a g c t g t t c t g g c c g a a

7440 145 140 135 130 MRTRAREIRHKAQHDFPCTA NeoR/KanR 145 140 135 130 MRTRAREIRHKAQHDFPCTA NeoR/KanR MRTRAREIRHKAQHDFPCTA c c a t c c g a g t a c g t g c t c t c t c g a t g c g a t g t t t c g c t t g g t g g t c g a a t g g g c a g g t a g g g t a g g c t c a t g c a c g a g a g a g c t a c g c t a c a a a g c g a a c c a c c a g c t t a c c c g t c c a t c

7500 125 120 115 110 PDLTHLRRMADAMISVKEAP NeoR/KanR 125 120 115 110 PDLTHLRRMADAMISVKEAP NeoR/KanR PDLTHLRRMADAMISVKEAP c c g g a t c a a g c g t a t g c a g c c g c c g c a t t g c a t c a g c c a t g a t g g a t a c t t t c t c g g c a g g g c c t a g t t c g c a t a c g t c g g c g g c g t a a c g t a g t c g g t a c t a c c t a t g a a a g a g c c g t c

7560 105 100 95 90 ALHSSLLDQGPVEGLLLWDR NeoR/KanR 105 100 95 90 ALHSSLLDQGPVEGLLLWDR NeoR/KanR ALHSSLLDQGPVEGLLLWDR g a g c a a g g t g a g a t g a c a g g a g a t c c t g c c c c g g c a c t t c g c c c a a t a g c a g c c a g t c c c c t c g t t c c a c t c t a c t g t c c t c t a g g a c g g g g c c g t g a a g c g g g t t a t c g t c g g t c a g g g

7620 85 80 75 70 GAETVVDLVAACPVGTTALW NeoR/KanR 85 80 75 70 GAETVVDLVAACPVGTTALW NeoR/KanR GAETVVDLVAACPVGTTALW t t c c c g c t t c a g t g a c a a c g t c g a g t a c a g c t g c g c a a g g a a c g c c c g t c g t g g c c a g c c a a g g g c g a a g t c a c t g t t g c a g c t c a t g t c g a c g c g t t c c t t g c g g g c a g c a c c g g t c g g

7680 65 60 55 50 SLRAAEDQLENLAGSLDTKV NeoR/KanR 65 60 55 50 SLRAAEDQLENLAGSLDTKV NeoR/KanR SLRAAEDQLENLAGSLDTKV a c g a t a g c c g c g c t g c c t c g t c t t g c a g t t c a t t c a g g g c a c c g g a c a g g t c g g t c t t g a t g c t a t c g g c g c g a c g g a g c a g a a c g t c a a g t a a g t c c c g t g g c c t g t c c a g c c a g a a c t

KasINarISfoIPluTI 7740 45 40 35 30 FLVPRGQASLRFVAADSCGI NeoR/KanR 45 40 35 30 FLVPRGQASLRFVAADSCGI NeoR/KanR FLVPRGQASLRFVAADSCGI c a a a a a g a a c c g g g c g c c c c t g c g c t g a c a g c c g g a a c a c g g c g g c a t c a g a g c a g c c g a g t t t t t c t t g g c c c g c g g g g a c g c g a c t g t c g g c c t t g t g c c g c c g t a g t c t c g t c g g c t

7800 25 20 15 10 TQQAWDYGFLREVWAAPSGA NeoR/KanR 25 20 15 10 TQQAWDYGFLREVWAAPSGA NeoR/KanR G C C C A G T C A T A G C C G A A T A G Neo-R TQQAWDYGFLREVWAAPSGA t t g t c t g t t g t g c c c a g t c a t a g c c g a a t a g c c t c t c c a c c c a a g c g g c c g g a g a a c c t g a a c a g a c a a c a c g g g t c a g t a t c g g c t t a t c g g a g a g g t g g g t t c g c c g g c c t c t t g g a c

7860 5 1 HLGDQEIM NeoR/KanR 5 1 HLGDQEIM NeoR/KanR HLGDQEIMINNFCKDPNTED c g t g c a a t c c a t c t t g t t c a a t c a t a a t a t t a t t g a a g c a t t t a t c a g g g t t c g t c t c g t g c a c g t t a g g t a g a a c a a g t t a g t a t t a t a a t a a c t t c g t a a a t a g t c c c a a g c a g a g c a

NsiI 3ʹ 5ʹ 7881 RDGGM c c c g g t c t c c t c c c a t g c a t g g g g c c a g a g g a g g g t a c g t 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.