Addgene pCSDest2-2XNLS-SpCas9-WT-NLS-3XHA-NLS-TALentry - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pCSDest2-2XNLS-SpCas9-WT-NLS-3XHA-NLS-TALentry


Map View

SP6 (37 .. 54) NdeI (8231) F1ori-R (7501 .. 7520) DraIII (7376) F1ori-F (7289 .. 7310) AmpR promoter Amp-R (6784 .. 6803) AhdI (6234) pBR322ori-F (5482 .. 5501) L4440 (5231 .. 5248) BspQI - SapI (5225) CAP binding site M13/pUC Reverse (5013 .. 5035) lac operator M13 Reverse (5000 .. 5016) M13 rev T3 (4961 .. 4981) T3 promoter NsiI (4934) SacII (4922) HpaI (4848) SV40pA-R (4792 .. 4811) EBV-rev (4738 .. 4757) SnaBI (4710) attB2 NLS 3xHA BstZ17I (4430) SV40 NLS SgrAI (4401) BspDI - ClaI (89) attB1 NcoI (142) Kozak sequence NLS NheI (195) BmtI (199) TspMI - XmaI (207) SmaI (209) BsiWI (237) BstAPI (249) NLS SbfI (541) XcmI (645) BlpI (1242) Bsu36I (1560) Eco53kI (1862) SacI (1864) AgeI (2785) BmgBI (2804) PmlI (3083) FspAI (3244) NruI (3534) PaqCI (3554) AscI (4249) pCSDest2-2XNLS-SpCas9-WT-NLS-3XHA-NLS-TALentry 8663 bp

Sequence View

5ʹ 3ʹ 60 CMV IE94 promoter SP6 promoter A T T T A G G T G A C A C T A T A G SP6 a c c g c c a t t c t g c c t g g g g a c g t c g g a g c a a g c t t g a t t t a g g t g a c a c t a t a g a a t a c a t g g c g g t a a g a c g g a c c c c t g c a g c c t c g t t c g a a c t a a a t c c a c t g t g a t a t c t t a t g t

BspDIClaI 120 attB1 a g c t a c t t g t t c t t t t t g c a g g a t c c c a t c g a t t c g a a t t c a a g g a t c a a c a a g t t t g t a t c g a t g a a c a a g a a a a a c g t c c t a g g g t a g c t a a g c t t a a g t t c c t a g t t g t t c a a a c a t

NcoI 180 1 5 PAAKKKK attB1 Kozak sequence NLS MGRAAPAAKKKK c a a a a a a g c a g g c t g g c g c c a c c a t g g g t a g a g c c g c g c c g g c a g c t a a g a a a a a g a a a c g t t t t t t c g t c c g a c c g c g g t g g t a c c c a t c t c g g c g c g g c c g t c g a t t c t t t t t c t t t g

BsiWITspMIXmaISmaIBmtINheI 240 LD (in frame with NLS) QLGASGSPGSGGSGSSGR NLS LDQLGASGSPGSGGSGSSGR *TAT t g g a t c a a t t g g g c g c t a g c g g t a g t c c c g g g t c t g g a g g t a g c g g c t c a a g t g g c c g t a a c c t a g t t a a c c c g c g a t c g c c a t c a g g g c c c a g a c c t c c a t c g c c g a g t t c a c c g g c a t

BstAPI 300 D Cas9 D Cas9 TAA 1 5 PAAKKKKLD (in frame with NLS) SEFGSGS D NLS Cas9 TAAPAAKKKKLDSEFGSGSD RCSRRCFLFLQVRFESASAV c g g c a g c t c c t g c g g c a a a a a a g a a a a a g t t g g a c t c t g a a t t c g g a a g c g g a a g c g a c a g c c g t c g a g g a c g c c g t t t t t t c t t t t t c a a c c t g a g a c t t a a g c c t t c g c c t t c g c t g t

360 5 10 15 20 KKYSIGLDIGTNSVGWAVIT Cas9 5 10 15 20 KKYSIGLDIGTNSVGWAVIT Cas9 5 10 15 20 KKYSIGLDIGTNSVGWAVIT Cas9 KKYSIGLDIGTNSVGWAVIT LLVGNPEIDTGVADAPGDNR a g a a g t 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 t c t t c a 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

420 25 30 35 40 DEYKVPSKKFKVLGNTDRHS Cas9 25 30 35 40 DEYKVPSKKFKVLGNTDRHS Cas9 25 30 35 40 DEYKVPSKKFKVLGNTDRHS Cas9 DEYKVPSKKFKVLGNTDRHS VLVLHRAFFEFNQAIGIAVA a c g a g t 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 g c t c 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

480 45 50 55 60 IKKNLIGALLFDSGETAEAT Cas9 45 50 55 60 IKKNLIGALLFDSGETAEAT Cas9 45 50 55 60 IKKNLIGALLFDSGETAEAT Cas9 IKKNLIGALLFDSGETAEAT YLLVENSGEQEVGPLRGFGR t a a a g a 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 a t t t c t 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

540 65 70 75 80 RLKRTARRRYTRRKNRICYL Cas9 65 70 75 80 RLKRTARRRYTRRKNRICYL Cas9 65 70 75 80 RLKRTARRRYTRRKNRICYL Cas9 RLKRTARRRYTRRKNRICYL PEFSCCPASIGASLIPDAVQ g g c t c a 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 c c g a g t 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

SbfI 600 85 90 95 100 QEIFSNEMAKVDDSFFHRLE Cas9 85 90 95 100 QEIFSNEMAKVDDSFFHRLE Cas9 85 90 95 100 QEIFSNEMAKVDDSFFHRLE Cas9 QEIFSNEMAKVDDSFFHRLE LLDKTILHSLHIVREEMPQL a g g a g a 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 t c c t 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

XcmI 660 105 110 115 120 ESFLVEEDKKHERHPIFGNI Cas9 105 110 115 120 ESFLVEEDKKHERHPIFGNI Cas9 105 110 115 120 ESFLVEEDKKHERHPIFGNI Cas9 ESFLVEEDKKHERHPIFGNI LGKQHLLIFLVLAVWDKAID a g t c c t 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 c a g g a 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

720 125 130 135 140 VDEVAYHEKYPTIYHLRKKL Cas9 125 130 135 140 VDEVAYHEKYPTIYHLRKKL Cas9 125 130 135 140 VDEVAYHEKYPTIYHLRKKL Cas9 VDEVAYHEKYPTIYHLRKKL HVLHRVMFLVWGYIM t g g a c g 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 a c c t g c 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

780 145 150 155 160 VDSTDKADLRLIYLALAHMI Cas9 145 150 155 160 VDSTDKADLRLIYLALAHMI Cas9 145 150 155 160 VDSTDKADLRLIYLALAHMI Cas9 VDSTDKADLRLIYLALAHMI t a g a c a 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 t c t g t 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

840 165 170 175 180 KFRGHFLIEGDLNPDNSDVD Cas9 165 170 175 180 KFRGHFLIEGDLNPDNSDVD Cas9 165 170 175 180 KFRGHFLIEGDLNPDNSDVD Cas9 KFRGHFLIEGDLNPDNSDVD a a t t t c 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 t t a a a g 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

900 185 190 195 200 KLFIQLVQTYNQLFEENPIN Cas9 185 190 195 200 KLFIQLVQTYNQLFEENPIN Cas9 185 190 195 200 KLFIQLVQTYNQLFEENPIN Cas9 KLFIQLVQTYNQLFEENPIN a a c t c t 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 t t g a g a 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

960 205 210 215 220 ASGVDAKAILSARLSKSRRL Cas9 205 210 215 220 ASGVDAKAILSARLSKSRRL Cas9 205 210 215 220 ASGVDAKAILSARLSKSRRL Cas9 ASGVDAKAILSARLSKSRRL c a t c c g 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 g t a g g 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

1020 225 230 235 240 ENLIAQLPGEKKNGLFGNLI Cas9 225 230 235 240 ENLIAQLPGEKKNGLFGNLI Cas9 225 230 235 240 ENLIAQLPGEKKNGLFGNLI Cas9 ENLIAQLPGEKKNGLFGNLI a a a a c c 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 t t t t g g 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

1080 245 250 255 260 ALSLGLTPNFKSNFDLAEDA Cas9 245 250 255 260 ALSLGLTPNFKSNFDLAEDA Cas9 245 250 255 260 ALSLGLTPNFKSNFDLAEDA Cas9 ALSLGLTPNFKSNFDLAEDA c c c t g t 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 g g g a c a 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

1140 265 270 275 280 KLQLSKDTYDDDLDNLLAQI Cas9 265 270 275 280 KLQLSKDTYDDDLDNLLAQI Cas9 265 270 275 280 KLQLSKDTYDDDLDNLLAQI Cas9 KLQLSKDTYDDDLDNLLAQI a g c t g c 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 t c g a c g 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

1200 285 290 295 300 GDQYADLFLAAKNLSDAILL Cas9 285 290 295 300 GDQYADLFLAAKNLSDAILL Cas9 285 290 295 300 GDQYADLFLAAKNLSDAILL Cas9 GDQYADLFLAAKNLSDAILL g c g a c c 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 c g c t g g 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

BlpI 1260 305 310 315 320 SDILRVNTEITKAPLSASMI Cas9 305 310 315 320 SDILRVNTEITKAPLSASMI Cas9 305 310 315 320 SDILRVNTEITKAPLSASMI Cas9 SDILRVNTEITKAPLSASMI g t g a t a 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 c a c t a 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

1320 325 330 335 340 KRYDEHHQDLTLLKALVRQQ Cas9 325 330 335 340 KRYDEHHQDLTLLKALVRQQ Cas9 325 330 335 340 KRYDEHHQDLTLLKALVRQQ Cas9 KRYDEHHQDLTLLKALVRQQ a g c g c t 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 c g c g a 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

1380 345 350 355 360 LPEKYKEIFFDQSKNGYAGY Cas9 345 350 355 360 LPEKYKEIFFDQSKNGYAGY Cas9 345 350 355 360 LPEKYKEIFFDQSKNGYAGY Cas9 LPEKYKEIFFDQSKNGYAGY t g c c t g 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 a c g g a c 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

1440 365 370 375 380 IDGGASQEEFYKFIKPILEK Cas9 365 370 375 380 IDGGASQEEFYKFIKPILEK Cas9 365 370 375 380 IDGGASQEEFYKFIKPILEK Cas9 IDGGASQEEFYKFIKPILEK t t g a c g 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 a a c t g c 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

1500 385 390 395 400 MDGTEELLVKLNREDLLRKQ Cas9 385 390 395 400 MDGTEELLVKLNREDLLRKQ Cas9 385 390 395 400 MDGTEELLVKLNREDLLRKQ Cas9 MDGTEELLVKLNREDLLRKQ t g g a c g 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 a c c t g c 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

1560 405 410 415 420 RTFDNGSIPHQIHLGELHAI Cas9 405 410 415 420 RTFDNGSIPHQIHLGELHAI Cas9 405 410 415 420 RTFDNGSIPHQIHLGELHAI Cas9 RTFDNGSIPHQIHLGELHAI g c a c t t 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 c g t g a a 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

Bsu36I 1620 425 430 435 440 LRRQEDFYPFLKDNREKIEK Cas9 425 430 435 440 LRRQEDFYPFLKDNREKIEK Cas9 425 430 435 440 LRRQEDFYPFLKDNREKIEK Cas9 LRRQEDFYPFLKDNREKIEK t c a g g c 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 a g t c c g 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

1680 445 450 455 460 ILTFRIPYYVGPLARGNSRF Cas9 445 450 455 460 ILTFRIPYYVGPLARGNSRF Cas9 445 450 455 460 ILTFRIPYYVGPLARGNSRF Cas9 ILTFRIPYYVGPLARGNSRF t c c t c a 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 a g g a g t 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

1740 465 470 475 480 AWMTRKSEETITPWNFEEVV Cas9 465 470 475 480 AWMTRKSEETITPWNFEEVV Cas9 465 470 475 480 AWMTRKSEETITPWNFEEVV Cas9 AWMTRKSEETITPWNFEEVV *FLGDSGPVELFDH c g t g g a 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 g c a c c t 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

1800 485 490 495 500 DKGASAQSFIERMTNFDKNL Cas9 485 490 495 500 DKGASAQSFIERMTNFDKNL Cas9 485 490 495 500 DKGASAQSFIERMTNFDKNL Cas9 DKGASAQSFIERMTNFDKNL ILPGRGLGEDFPHSVKIFIQ a t a a g g 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 t a t t c c 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

1860 505 510 515 520 PNEKVLPKHSLLYEYFTVYN Cas9 505 510 515 520 PNEKVLPKHSLLYEYFTVYN Cas9 505 510 515 520 PNEKVLPKHSLLYEYFTVYN Cas9 PNEKVLPKHSLLYEYFTVYN RVFLHKRFVRQQVLVECNIV c t a a c g 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 g a t t g c 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

Eco53kISacI 1920 525 530 535 540 ELTKVKYVTEGMRKPAFLSG Cas9 525 530 535 540 ELTKVKYVTEGMRKPAFLSG Cas9 525 530 535 540 ELTKVKYVTEGMRKPAFLSG Cas9 ELTKVKYVTEGMRKPAFLSG LEGLDFVDCFPHSLWCEQRS a g c t c a 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 t c g a g t 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

1980 545 550 555 560 EQKKAIVDLLFKTNRKVTVK Cas9 545 550 555 560 EQKKAIVDLLFKTNRKVTVK Cas9 545 550 555 560 EQKKAIVDLLFKTNRKVTVK Cas9 EQKKAIVDLLFKTNRKVTVK LLLFSDHVEEELRVPFNGHF a g c a g a 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 t c g t c t 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

2040 565 570 575 580 QLKEDYFKKIECFDSVEISG Cas9 565 570 575 580 QLKEDYFKKIECFDSVEISG Cas9 565 570 575 580 QLKEDYFKKIECFDSVEISG Cas9 QLKEDYFKKIECFDSVEISG LEFLVIEFLNFTEVRNFDAS a g c t c a a a g a g 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 c g a g t t t c t c 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

2100 585 590 595 600 VEDRFNASLGTYHDLLKIIK Cas9 585 590 595 600 VEDRFNASLGTYHDLLKIIK Cas9 585 590 595 600 VEDRFNASLGTYHDLLKIIK Cas9 VEDRFNASLGTYHDLLKIIK HLIAEVCGQSRIVIEQFDNF t g g a g g 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 c t c c 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

2160 605 610 615 620 DKDFLDNEENEDILEDIVLT Cas9 605 610 615 620 DKDFLDNEENEDILEDIVLT Cas9 605 610 615 620 DKDFLDNEENEDILEDIVLT Cas9 DKDFLDNEENEDILEDIVLT VLVEQVILLVLVNKLVNDEG 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 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 g t t c c 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

2220 625 630 635 640 LTLFEDREMIEERLKTYAHL Cas9 625 630 635 640 LTLFEDREMIEERLKTYAHL Cas9 625 630 635 640 LTLFEDREMIEERLKTYAHL Cas9 LTLFEDREMIEERLKTYAHL KRQKFIPLHNFFAQFSVSM t t a c g t 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 a a t g c a 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

2280 645 650 655 660 FDDKVMKQLKRRRYTGWGRL Cas9 645 650 655 660 FDDKVMKQLKRRRYTGWGRL Cas9 645 650 655 660 FDDKVMKQLKRRRYTGWGRL Cas9 FDDKVMKQLKRRRYTGWGRL t c g a c g 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 a g c t g c 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

2340 665 670 675 680 SRKLINGIRDKQSGKTILDF Cas9 665 670 675 680 SRKLINGIRDKQSGKTILDF Cas9 665 670 675 680 SRKLINGIRDKQSGKTILDF Cas9 SRKLINGIRDKQSGKTILDF *SFQDIADSVLLTSLCDQIK c a a g a a 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 g t t c t 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

2400 685 690 695 700 LKSDGFANRNFMQLIHDDSL Cas9 685 690 695 700 LKSDGFANRNFMQLIHDDSL Cas9 685 690 695 700 LKSDGFANRNFMQLIHDDSL Cas9 LKSDGFANRNFMQLIHDDSL KLGISKGVPVEHLQDMIVRE t t a a g t 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 a a t t c 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

2460 705 710 715 720 TFKEDIQKAQVSGQGDSLHE Cas9 705 710 715 720 TFKEDIQKAQVSGQGDSLHE Cas9 705 710 715 720 TFKEDIQKAQVSGQGDSLHE Cas9 TFKEDIQKAQVSGQGDSLHE GKLLVDLFCLNRALPVTEVL c c t t t a 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 c c a c g a g c g g a a a t 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 g g t g c t c g

2520 725 730 735 740 HIANLAGSPAIKKGILQTVK Cas9 725 730 735 740 HIANLAGSPAIKKGILQTVK Cas9 725 730 735 740 HIANLAGSPAIKKGILQTVK Cas9 HIANLAGSPAIKKGILQTVK VDSIKCTAWSDFLSYQLGNL a c a t c g 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 g t a g c 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

2580 745 750 755 760 VVDELVKVMGRHKPENIVIE Cas9 745 750 755 760 VVDELVKVMGRHKPENIVIE Cas9 745 750 755 760 VVDELVKVMGRHKPENIVIE Cas9 VVDELVKVMGRHKPENIVIE DHIFEDFYHSPMLGLIDNDL t c g t g g 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 a g c a 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

2640 765 770 775 780 MARENQTTQKGQKNSRERMK Cas9 765 770 775 780 MARENQTTQKGQKNSRERMK Cas9 765 770 775 780 MARENQTTQKGQKNSRERMK Cas9 MARENQTTQKGQKNSRERMK HGSLVLSGLLSLLVTPFPHL t g g c c c 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 a c c g g 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

2700 785 790 795 800 RIEEGIKELGSQILKEHPVE Cas9 785 790 795 800 RIEEGIKELGSQILKEHPVE Cas9 785 790 795 800 RIEEGIKELGSQILKEHPVE Cas9 RIEEGIKELGSQILKEHPVE PNFLTYFFQPGLDKLFVWNF g g a t t g 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 c c t a a 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

2760 805 810 815 820 NTQLQNEKLYLYYLQNGRDM Cas9 805 810 815 820 NTQLQNEKLYLYYLQNGRDM Cas9 805 810 815 820 NTQLQNEKLYLYYLQNGRDM Cas9 NTQLQNEKLYLYYLQNGRDM VGLKLILLEVQVVQLVAPVH a c a c c c 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 t g t g 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

BmgBIAgeI 2820 825 830 835 840 YVDQELDINRLSDYDVDHIV Cas9 825 830 835 840 YVDQELDINRLSDYDVDHIV Cas9 825 830 835 840 YVDQELDINRLSDYDVDHIV Cas9 YVDQELDINRLSDYDVDHIV VHILFQVDVPQGVVVHIM a c g t g g 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 t g c a c c 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

2880 845 850 855 860 PQSFLKDDSIDNKVLTRSDK Cas9 845 850 855 860 PQSFLKDDSIDNKVLTRSDK Cas9 845 850 855 860 PQSFLKDDSIDNKVLTRSDK Cas9 PQSFLKDDSIDNKVLTRSDK c c c a a a 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 g g g t t t 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

2940 865 870 875 880 NRGKSDNVPSEEVVKKMKNY Cas9 865 870 875 880 NRGKSDNVPSEEVVKKMKNY Cas9 865 870 875 880 NRGKSDNVPSEEVVKKMKNY Cas9 NRGKSDNVPSEEVVKKMKNY a t a g a g 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 t a t c t 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

3000 885 890 895 900 WRQLLNAKLITQRKFDNLTK Cas9 885 890 895 900 WRQLLNAKLITQRKFDNLTK Cas9 885 890 895 900 WRQLLNAKLITQRKFDNLTK Cas9 WRQLLNAKLITQRKFDNLTK g g c g g c 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 c g c c g 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

3060 905 910 915 920 AERGGLSELDKAGFIKRQLV Cas9 905 910 915 920 AERGGLSELDKAGFIKRQLV Cas9 905 910 915 920 AERGGLSELDKAGFIKRQLV Cas9 AERGGLSELDKAGFIKRQLV c t g a a c 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 g a c t t g 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

PmlI 3120 925 930 935 940 ETRQITKHVAQILDSRMNTK Cas9 925 930 935 940 ETRQITKHVAQILDSRMNTK Cas9 925 930 935 940 ETRQITKHVAQILDSRMNTK Cas9 ETRQITKHVAQILDSRMNTK a g a c a c 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 t c t g 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

3180 945 950 955 960 YDENDKLIREVKVITLKSKL Cas9 945 950 955 960 YDENDKLIREVKVITLKSKL Cas9 945 950 955 960 YDENDKLIREVKVITLKSKL Cas9 YDENDKLIREVKVITLKSKL a c g a t g 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 g c t a c 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

3240 965 970 975 980 VSDFRKDFQFYKVREINNYH Cas9 965 970 975 980 VSDFRKDFQFYKVREINNYH Cas9 965 970 975 980 VSDFRKDFQFYKVREINNYH Cas9 VSDFRKDFQFYKVREINNYH *IESLVKLKILHSLDVIVV t c t c a g 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 g a g t 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

FspAI 3300 985 990 995 1000 HAHDAYLNAVVGTALIKKYP Cas9 985 990 995 1000 HAHDAYLNAVVGTALIKKYP Cas9 985 990 995 1000 HAHDAYLNAVVGTALIKKYP Cas9 HAHDAYLNAVVGTALIKKYP MRMIGVQICHYASCKDFFIG a t g c g c 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 t a c g c 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

3360 1005 1010 1015 1020 KLESEFVYGDYKVYDVRKMI Cas9 1005 1010 1015 1020 KLESEFVYGDYKVYDVRKMI Cas9 1005 1010 1015 1020 KLESEFVYGDYKVYDVRKMI Cas9 KLESEFVYGDYKVYDVRKMI LQFRFKNVSVIFHVINPFHD a g c t g g 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 t c g a c c 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

3420 1025 1030 1035 1040 AKSEQEIGKATAKYFFYSNI Cas9 1025 1030 1035 1040 AKSEQEIGKATAKYFFYSNI Cas9 1025 1030 1035 1040 AKSEQEIGKATAKYFFYSNI Cas9 AKSEQEIGKATAKYFFYSNI CLRLLFYALGGSLVEKVAIN c a a a g t 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 g t t t c a 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

3480 1045 1050 1055 1060 MNFFKTEITLANGEIRKRPL Cas9 1045 1050 1055 1060 MNFFKTEITLANGEIRKRPL Cas9 1045 1050 1055 1060 MNFFKTEITLANGEIRKRPL Cas9 MNFFKTEITLANGEIRKRPL HIKELGLNCQGISLNPLSWK t g a a t t 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 a c t t 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

NruI 3540 1065 1070 1075 1080 IETNGETGEIVWDKGRDFAT Cas9 1065 1070 1075 1080 IETNGETGEIVWDKGRDFAT Cas9 1065 1070 1075 1080 IETNGETGEIVWDKGRDFAT Cas9 IETNGETGEIVWDKGRDFAT DFCVSFCSFDHPVLTPIERC t c g a a a 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 a g c t t t 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

PaqCI 3600 1085 1090 1095 1100 VRKVLSMPQVNIVKKTEVQT Cas9 1085 1090 1095 1100 VRKVLSMPQVNIVKKTEVQT Cas9 1085 1090 1095 1100 VRKVLSMPQVNIVKKTEVQT Cas9 VRKVLSMPQVNIVKKTEVQT DALDQGHRLHVDNFLGFYLG t c c g c a 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 a g g c g 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

3660 1105 1110 1115 1120 GGFSKESILPKRNSDKLIAR Cas9 1105 1110 1115 1120 GGFSKESILPKRNSDKLIAR Cas9 1105 1110 1115 1120 GGFSKESILPKRNSDKLIAR Cas9 GGFSKESILPKRNSDKLIAR SAEGLFTDERFPVAVLQDCA g a g g c t 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 c t c c g a 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

3720 1125 1130 1135 1140 KKDWDPKKYGGFDSPTVAYS Cas9 1125 1130 1135 1140 KKDWDPKKYGGFDSPTVAYS Cas9 1125 1130 1135 1140 KKDWDPKKYGGFDSPTVAYS Cas9 KKDWDPKKYGGFDSPTVAYS FFIPVGLFVASEIRRCDSVT a a a a a g 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 t t t t c 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

3780 1145 1150 1155 1160 VLVVAKVEKGKSKKLKSVKE Cas9 1145 1150 1155 1160 VLVVAKVEKGKSKKLKSVKE Cas9 1145 1150 1155 1160 VLVVAKVEKGKSKKLKSVKE Cas9 VLVVAKVEKGKSKKLKSVKE YQNHGFHLFPLRFFEFADLF t a c t g g 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 a t g a c c 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

3840 1165 1170 1175 1180 LLGITIMERSSFEKNPIDFL Cas9 1165 1170 1175 1180 LLGITIMERSSFEKNPIDFL Cas9 1165 1170 1175 1180 LLGITIMERSSFEKNPIDFL Cas9 LLGITIMERSSFEKNPIDFL QQADCDHLSGAELFVGDVKE t g c t g g 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 c g a c c 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

3900 1185 1190 1195 1200 EAKGYKEVKKDLIIKLPKYS Cas9 1185 1190 1195 1200 EAKGYKEVKKDLIIKLPKYS Cas9 1185 1190 1195 1200 EAKGYKEVKKDLIIKLPKYS Cas9 EAKGYKEVKKDLIIKLPKYS FRFSIFLDFFVEDNLQGLVR a a g c g a 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 t c g c t 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

3960 1205 1210 1215 1220 LFELENGRKRMLASAGELQK Cas9 1205 1210 1215 1220 LFELENGRKRMLASAGELQK Cas9 1205 1210 1215 1220 LFELENGRKRMLASAGELQK Cas9 LFELENGRKRMLASAGELQK EKLKFVAPFSHESTRALQLF t c t t t g 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 a g a a a c 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

4020 1225 1230 1235 1240 GNELALPSKYVNFLYLASHY Cas9 1225 1230 1235 1240 GNELALPSKYVNFLYLASHY Cas9 1225 1230 1235 1240 GNELALPSKYVNFLYLASHY Cas9 GNELALPSKYVNFLYLASHY TVLQCQGRFVNIEQIQGAVI g t a a c g 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 c a t t 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

4080 1245 1250 1255 1260 EKLKGSPEDNEQKQLFVEQH Cas9 1245 1250 1255 1260 EKLKGSPEDNEQKQLFVEQH Cas9 1245 1250 1255 1260 EKLKGSPEDNEQKQLFVEQH Cas9 EKLKGSPEDNEQKQLFVEQH FLEFPRGFIILLLLQEHFLV a a a a g c 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 t t t t c g 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

4140 1265 1270 1275 1280 KHYLDEIIEQISEFSKRVIL Cas9 1265 1270 1275 1280 KHYLDEIIEQISEFSKRVIL Cas9 1265 1270 1275 1280 KHYLDEIIEQISEFSKRVIL Cas9 KHYLDEIIEQISEFSKRVIL FVVKILDDLLYALEGFSHDE a a c a c t 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 t t g t g a 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

4200 1285 1290 1295 1300 ADANLDKVLSAYNKHRDKPI Cas9 1285 1290 1295 1300 ADANLDKVLSAYNKHRDKPI Cas9 1285 1290 1295 1300 ADANLDKVLSAYNKHRDKPI Cas9 ADANLDKVLSAYNKHRDKPI GVSVEILHKRSVILVPILGD c c g a c g 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 c t 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

AscI 4260 1305 1310 1315 1320 REQAENIIHLFTLTNLGAPA Cas9 1305 1310 1315 1320 REQAENIIHLFTLTNLGAPA Cas9 1305 1310 1315 1320 REQAENIIHLFTLTNLGAPA Cas9 REQAENIIHLFTLTNLGAPA PLLCFVNDVQKSQGVQARRC g g g a g c 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 c t c g 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

4320 1325 1330 1335 1340 AFKYFDTTIDRKRYTSTKEV Cas9 1325 1330 1335 1340 AFKYFDTTIDRKRYTSTKEV Cas9 1325 1330 1335 1340 AFKYFDTTIDRKRYTSTKEV Cas9 AFKYFDTTIDRKRYTSTKEV GELVEVGGYVSLPVGRCLLD c c t t c a 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 g g a a g t 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

4380 1345 1350 1355 1360 LDATLIHQSITGLYETRIDL Cas9 1345 1350 1355 1360 LDATLIHQSITGLYETRIDL Cas9 1345 1350 1355 1360 LDATLIHQSITGLYETRIDL Cas9 LDATLIHQSITGLYETRIDL QVGCQNM t g g a c g 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 a c c t 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

BstZ17ISgrAI 4440 1365 SQLGGD (in frame with Cas9) GTGGPKKKRKVYPY Cas9 1365 SQLGGD (in frame with Cas9) GTGGPKKKRKVYPY Cas9 1365 SQLGGD GTGG 130 PKKKRKV 1 YPY Cas9 SV40 NLS 3xHA SQLGGDGTGGPKKKRKVYPY c t c a g c 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 g a g t c g 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

4500 (in frame with Cas9) DVPDYAGYPYDVPDYAGSYP (in frame with Cas9) DVPDYAGYPYDVPDYAGSYP 5 10 15 20 DVPDYAGYPYDVPDYAGSYP 3xHA DVPDYAGYPYDVPDYAGSYP a t g t t c 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 t a c a a 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

4560 (in frame with Cas9) YDVPDYAGSAAPAAKKKKLD (in frame with Cas9) YDVPDYAGSAAPAAKKKKLD 25 30 YDVPDYA GSAA 1 5 PAAKKKKLD 3xHA NLS YDVPDYAGSAAPAAKKKKLD a t g a c g 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 a c t g c 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

4620 (in frame with Cas9) FESGVQVFMLV* (in frame with Cas9) FESGVQVFMLV* (in frame with NLS) FESGVQVFMLV* FESGVQVFMLV* t c g a a t c c g g a g t a c a a g t c t t c a t g c t a g t a t a g c t g a a g a c a a g a t c t t a a t c c g g a t a g c t t a g g c c t c a t g t t c a g a a g t a c g a t c a t a t c g a c t t c t g t t c t a g a a t t a g g c c t a

4680 attB2 a a g c g g c c g c c t c g a g a t a t c t a g a c c c a g c t t t c t t g t a c a a a g t g g t t g a t c c t c t c g t t c g c c g g c g g a g c t c t a t a g a t c t g g g t c g a a a g a a c a t g t t t c a c c a a c t a g g a g a g c

SnaBI 4740 SV40 poly(A) signal C T A EBV-rev a g c c t c t a g a a c t a t a g t g a g t c g t a t t a c g t a g a t c c a g a c a t g a t a a g a t a c a t t g a t t c g g a g a t c t t g a t a t c a c t c a g c a t a a t g c a t c t a g g t c t g t a c t a t t c t a t g t a a c t a

4800 SV40 poly(A) signal G A A A T T T G T SV40pA-R C T C A A A C C T G T T T G G T G EBV-rev g a g t 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 c t c a 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

HpaI 4860 SV40 poly(A) signal G A T G C T A T T G C SV40pA-R g a t g 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 c t a c 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

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

NsiISacII 4980 T3 promoter G G A A A T C A C T C C C A A T T A A C T3 g c g g c g c c a a t g c a t t g g g c c c g g t a c 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 t g c g c c g c g g t t a c g t a a c c c g g g c c a t g 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 a c

5040 M13 rev lac operator G T3 C A G T A T C G A C A A A G G A C M13 Reverse G G A C A C A C T T T A A C A A T A G G C G A M13/pUC Reverse c g c g c t t g g c g t a a t c a t g g t c a t a g c t g t t t c c t g t g t g a a a t t g t t a t c c g c t c a c a a g c g c g a a c c g c a t t a g t a c c a g t a t c g a c a a a g g a c a c a c t t t a a c a a t a g g c g a g t g t t

5100 -10 -35 lac promoter CAP binding site t t c c a c a c a a c a t a c g a g c c g g a a g c a t a a a g t g t a a a g c c t g g g g t g c c t a a t g a g t g a a a g g t g t g t t g t a t g c t c g g c c t t c g t a t t t c a c a t t t c g g a c c c c a c g g a t t a c t c a c t

5160 CAP binding site g c t a a c t c a c a t t a a t t g c g t t g c g c t c a c t g c c c g c t t t c c a g t c g g g a a a c c t g t c g t c g a t t g a g t g t a a t t a a c g c a a c g c g a g t g a c g g g c g a a a g g t c a g c c c t t t g g a c a g c a

5220 g c c a g c t g c a t t a a t g a a t c g g c c a a c g c g c g g g g a g a g g c g g t t t g c g t a t t g g g c g c t c g g t c g a c g t a a t t a c t t a g c c g g t t g c g c g c c c c t c t c c g c c a a a c g c a t a a c c c g c g a

BspQISapI 5280 G A G C G A G T G A C T G A G C G A L4440 c t t c c g c t t c c t c g c t c a c t g a c t c g c t g c g c t c g g t c g t t c g g c t g c g g c g a g c g g t a t g a a g g c g a a g g a g c g a g t g a c t g a g c g a c g c g a g c c a g c a a g c c g a c g c c g c t c g c c a t a

5340 c a g c t c a c t c a a a g g c g g t a a t a c g g t t a t c c a c a g a a t c a g g g g a t a a c g c a g g a a a g a g t c g a g t g a g t t t c c g c c a t t a t g c c a a t a g g t g t c t t a g t c c c c t a t t g c g t c c t t t c t

5400 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

5460 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

5520 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

5580 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 c 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 g 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

5640 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

5700 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

5760 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

5820 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

5880 ori c t a a c t a c g g c t a c a c t a g a a g g 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 c 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

5940 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

6000 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

6060 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

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

6180 285 *WHKILS AmpR *WHKILS a a t c a a t c t a a a g t a t a t a t g a g t a a a c t t g g t c t g a c a g t t a c c a a t g c t t a a t c a g t g t t a g t t a g a t t t c a t a t a t a c t c a t t t g a a c c a g a c t g t c a a t g g t t a c g a a t t a g t c a c

AhdI 6240 280 275 270 265 AGIEAIQRNREDMTAQSGTT AmpR AGIEAIQRNREDMTAQSGTT a g g c a c c t a t c t c a g c g a t c t g t c t a t t t c g t t c a t c c a t a g t t g c c t g a c t c c c c g t c g t c c g t g g a t a g a g t c g c t a g a c a g a t a a a g c a a g t a g g t a t c a a c g g a c t g a g g g g c a g c

6300 260 255 250 245 YIVVIRSPKGDPGLAAIIGR AmpR YIVVIRSPKGDPGLAAIIGR t g t a g a t a a c t a c g a t a c g g g a g g g c t t a c c a t c t g g c c c c a g t g c t g c a a t g a t a c c g c a c a t c t a t t g a t g c t a t g c c c t c c c g a a t g g t a g a c c g g g g t c a c g a c g t t a c t a t g g c g

6360 240 235 230 225 SGREGAGSKDAIFWGAPLAS AmpR SGREGAGSKDAIFWGAPLAS g a g a c c c a c g c t c a c c g g c t c c a g a t t t a t c a g c a a t a a a c c a g c c a g c c g g a a g g g c c g c t c t g g g t g c g a g t g g c c g a g g t c t a a a t a g t c g t t a t t t g g t c g g t c g g c c t t c c c g g c

6420 220 215 210 205 RLLPGAVKDAEMWDILQQRS AmpR RLLPGAVKDAEMWDILQQRS a g c g c a g a a g t g g t c c t g c a a c t t t a t c c g c c t c c a t c c a g t c t a t t a a t t g t t g c c g g g t c g c g t c t t c a c c a g g a c g t t g a a a t a g g c g g a g g t a g g t c a g a t a a t t a a c a a c g g c c c

6480 200 195 190 185 ALTLLEGTLLKRLTTAMAVP AmpR ALTLLEGTLLKRLTTAMAVP a a g c t a g a g t a a g t a g t t c g c c a g t t a a t a g t t t g c g c a a c g t t g t t g c c a t t g c t a c a g t t c g a t c t c a t t c a t c a a g c g g t c a a t t a t c a a a c g c g t t g c a a c a a c g g t a a c g a t g t c

6540 180 175 170 165 MTTDREDNPIAENLEPEWRD AmpR MTTDREDNPIAENLEPEWRD g c a t c g t g g t g t c a c g c t c g t c g t t t g g t a t g g c t t c a t t c a g c t c c g g t t c c c a a c g a t c g t a g c a c c a c a g t g c g a g c a g c a a a c c a t a c c g a a g t a a g t c g a g g c c a a g g g t t g c t a

6600 160 155 150 145 LRTVHDGMNHLFATLEKPGG AmpR LRTVHDGMNHLFATLEKPGG c a a g g c g a g t t a c a t g a t c c c c c a t g t t g t g c a a a a a a g c g g t t a g c t c c t t c g g t c c t c g t t c c g c t c a a t g t a c t a g g g g g t a c a a c a c g t t t t t t c g c c a a t c g a g g a a g c c a g g a g

6660 140 135 130 125 ITTLLLNAATNDSMTIAASC AmpR ITTLLLNAATNDSMTIAASC c g a t c g t t g t c a g a a g t a a g t t g g c c g c a g t g t t a t c a c t c a t g g t t a t g g c a g c a c t g c g c t a g c a a c a g t c t t c a t t c a a c c g g c g t c a c a a t a g t g a g t a c c a a t a c c g t c g t g a c g

6720 120 115 110 105 LERVTMGDTLHKETVPSYEV AmpR LERVTMGDTLHKETVPSYEV a t a a t t c t c t t a c t g t c a t g c c a t c c g t a a g a t g c t t t t c t g t g a c t g g t g a g t a c t c a a t a t t a a g a g a a t g a c a g t a c g g t a g g c a t t c t a c g a a a a g a c a c t g a c c a c t c a t g a g t t

6780 100 95 90 85 LDNQSYHIRRGLQEQGADIR AmpR LDNQSYHIRRGLQEQGADIR c c a a g t c a t t c t g a g a a t a g t g t a t g c g g c g a c c g a g t t g c t c t t g c c c g g c g t c a a t a c g g t t c a g t a a g a c t c t t a t c a c a t a c g c c g c t g g c t c a a c g a g a a c g g g c c g c a g t t a t g

6840 80 75 70 65 SLVAGCLLVKFTSMMPFREE AmpR A T A A T A C C G C G C C A C A T A G C Amp-R SLVAGCLLVKFTSMMPFREE g g g a t a a t a c c g c g c c a c a t a g c a g a a c t t t a a a a g t g c t c a t c a t t g g a a a a c g t t c t t c c c t a t t a t g g c g c g g t g t a t c g t c t t g a a a t t t t c a c g a g t a g t a a c c t t t t g c a a g a a

6900 60 55 50 45 PRFSELIKGSNLDLEIYGVR AmpR PRFSELIKGSNLDLEIYGVR c g g g g c g a a a a c t c t c a a g g a t c t t a c c g c t g t t g a g a t c c a g t t c g a t g t a a c c c a c t c g c c c c g c t t t t g a g a g t t c c t a g a a t g g c g a c a a c t c t a g g t c a a g c t a c a t t g g g t g a g

6960 40 35 30 25 AGLQDEADKVKVLTEPH signal sequence AFV AmpR AGLQDEADKVKVLTEPHAFV g t g c a c c c a a c t g a t c t t c a g c a t c t t t t a c t t t c a c c a g c g t t t c t g g g t g a g c a a a a a c a c g t g g g t t g a c t a g a a g t c g t a g a a a a t g a a a g t g g t c g c a a a g a c c c a c t c g t t t t t

7020 20 15 10 5 1 signal sequence PLCFAAFFPILAVRFHQISM AmpR PLCFAAFFPILAVRFHQISM c a g g a a g g c a a a a t g c c g c a a a a a a g g g a a t a a g g g c g a c a c g g a a a t g t t g a a t a c t c a g t c c t t c c g t t t t a c g g c g t t t t t t c c c t t a t t c c c g c t g t g c c t t t a c a a c t t a t g a g t

7080 AmpR AmpR promoter t a c t c t t c c t t t t t c a a t a t t a t t g a a g c a t t t a t c a g g g t t a t t g t c t c a t g a g c g g a t a t g a g a a g g a a a a a g t t a t a a t a a c t t c g t a a a t a g t c c c a a t a a c a g a g t a c t c g c c t a

7140 AmpR promoter a c a t a t t t g a a t g t a t t t a g a a a a a t a a a c a a a t a g g g g t t c c g c g c a c a t t t c c c c g a a t g t a t a a a c t t a c a t a a a t c t t t t t a t t t g t t t a t c c c c a a g g c g c g t g t a a a g g g g c t t

7200 f1 ori a a g t g c c a c c t a a a t t g t a a g c g t t a a t a t t t t g t t a a a a t t c g c g t t a a a t t t t t g t t a t t c a c g g t g g a t t t a a c a t t c g c a a t t a t a a a a c a a t t t t a a g c g c a a t t t a a a a a c a a t

7260 f1 ori a a t c a g c t c a t t t t t t a a c c a a t a g g c c g a a a t c g g c a a a a t c c c t t a t a a a t c a a a a g a t t a g t c g a g t a a a a a a t t g g t t a t c c g g c t t t a g c c g t t t t a g g g a a t a t t t a g t t t t c t

7320 f1 ori G G T C A A A C C T T G T T C T C A G G T G F1ori-F a t a g a c c g a g a t a g g g t t g a g t g t t g t t c c a g t t t g g a a c a a g a g t c c a c t a t t a a a g a a t a t c t g g c t c t a t c c c a a c t c a c a a c a a g g t c a a a c c t t g t t c t c a g g t g a t a a t t t c t t

DraIII 7380 f1 ori c g t g g a c t c c a a c g t c a a a g g g c g a a a a a c c g t c t a t c a g g g c g a t g g c c c a c t a c g t g a g c a c c t g a g g t t g c a g t t t c c c g c t t t t t g g c a g a t a g t c c c g c t a c c g g g t g a t g c a c t

7440 f1 ori a c c a t c a c c c t a a t c a a g t t t t t t g g g g t c g a g g t g c c g t a a a g c a c t a a a t c g g a a c c c t g g t a g t g g g a t t a g t t c a a a a a a c c c c a g c t c c a c g g c a t t t c g t g a t t t a g c c t t g g g

7500 f1 ori t a a a g g g a g c c c c c g a t t t a g a g c t t g a c g g g g a a a g c c g g c g a a c g t g g c g a g a a a g g a a t t t c c c t c g g g g g c t a a a t c t c g a a c t g c c c c t t t c g g c c g c t t g c a c c g c t c t t t c c t

7560 f1 ori A G G G A A G A A A G C G A A A G G A G F1ori-R a g g g a a g a a a g c g a a a g g a g c g g g c g c t a g g g c g c t g g c a a g t g t a g c g g t c a c g c t g c g t c c c t t c t t t c g c t t t c c t c g c c c g c g a t c c c g c g a c c g t t c a c a t c g c c a g t g c g a c g c

7620 f1 ori c g t a a c c a c c a c a c c c g c c g c g c t t a a t g c g c c g c t a c a g g g c g c g t c c c a t t c g c c a t t g c a t t g g t g g t g t g g g c g g c g c g a a t t a c g c g g c g a t g t c c c g c g c a g g g t a a g c g g t a a

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

7740 CMV IE94 promoter g a c c a t a g c c a a t t c a a t a t g g c g t a t a t g g a c t c a t g c c a a t t c a a t a t g g t g g a t c t g c t g g t a t c g g t t a a g t t a t a c c g c a t a t a c c t g a g t a c g g t t a a g t t a t a c c a c c t a g a c

7800 CMV IE94 promoter g a c c t g t g c c a a t t c a a t a t g g c g t a t a t g g a c t c g t g c c a a t t c a a t a t g g t g g a t c t g c t g g a c a c g g t t a a g t t a t a c c g c a t a t a c c t g a g c a c g g t t a a g t t a t a c c a c c t a g a c

7860 CMV IE94 promoter g a c c c c a g c c a a t t c a a t a t g g c g g a c t t g g c a c c a t g c c a a t t c a a t a t g g c g g a c t t g c t g g g g t c g g t t a a g t t a t a c c g c c t g a a c c g t g g t a c g g t t a a g t t a t a c c g c c t g a a c

7920 CMV IE94 promoter g c a c t g t g c c a a c t g g g g a g g g g t c t a c t t g g c a c g g t g c c a a g t t t g a g g a g g g g t c t t c g t g a c a c g g t t g a c c c c t c c c c a g a t g a a c c g t g c c a c g g t t c a a a c t c c t c c c c a g a a

7980 CMV IE94 promoter g g c c c t g t g c c a a g t c c g c c a t a t t g a a t t g g c a t g g t g c c a a t a a t g g c g g c c a t a t t g c c g g g a c a c g g t t c a g g c g g t a t a a c t t a a c c g t a c c a c g g t t a t t a c c g c c g g t a t a a c

8040 CMV IE94 promoter g c t a t a t g c c a g g a t c a a t a t a t a g g c a a t a t c c a a t a t g g c c c t a t g c c a a t a t g g c t a c g a t a t a c g g t c c t a g t t a t a t a t c c g t t a t a g g t t a t a c c g g g a t a c g g t t a t a c c g a t

8100 CMV IE94 promoter t t g g c c a g g t t c a a t a c t a t g t a t t g g c c c t a t g c c a t a t a g t a t t c c a t a t a t g g g t t t a a c c g g t c c a a g t t a t g a t a c a t a a c c g g g a t a c g g t a t a t c a t a a g g t a t a t a c c c a a a

8160 CMV IE94 promoter t c c t a t t g a c g t a g a t a g c c c c t c c c a a t g g g c g g t c c c a t a t a c c a t a t a t g g g g c t t c a g g a t a a c t g c a t c t a t c g g g g a g g g t t a c c c g c c a g g g t a t a t g g t a t a t a c c c c g a a g

8220 CMV IE94 promoter c t a a t a c c g c c c a t a g c c a c t c c c c c a t t g a c g t c a a t g g t c t c t a t a t a t g g t c t t t c c g a t t a t g g c g g g t a t c g g t g a g g g g g t a a c t g c a g t t a c c a g a g a t a t a t a c c a g a a a g g

NdeI 8280 CMV IE94 promoter t a t t g a c g t c a t a t g g g c g g t c c t a t t g a c g t a t a t g g c g c c t c c c c c a t t g a c g t c a a t a t a a c t g c a g t a t a c c c g c c a g g a t a a c t g c a t a t a c c g c g g a g g g g g t a a c t g c a g t t a

8340 CMV IE94 promoter t a c g g t a a a t g g c c c g c c t g g c t c a a t g c c c a t t g a c g t c a a t a g g a c c a c c c a c c a t t g a t g c c a t t t a c c g g g c g g a c c g a g t t a c g g g t a a c t g c a g t t a t c c t g g t g g g t g g t a a c

8400 CMV IE94 promoter a c g t c a a t g g g a t g g c t c a t t g c c c a t t c a t a t c c g t t c t c a c g c c c c c t a t t g a c g t c a t g c a g t t a c c c t a c c g a g t a a c g g g t a a g t a t a g g c a a g a g t g c g g g g g a t a a c t g c a g t

8460 CMV IE94 promoter a t g a c g g t a a a t g g c c c a c t t g g c a g t a c a t c a a t a t c t a t t a a t a g t a a c t t g g c a a g t t a c t g c c a t t t a c c g g g t g a a c c g t c a t g t a g t t a t a g a t a a t t a t c a t t g a a c c g t t c a

8520 CMV IE94 promoter a c a t t a c t a t t g g a a g g a c g c c a g g g t a c a t t g g c a g t a c t c c c a t t g a c g t c a a t g g c g t g t a a t g a t a a c c t t c c t g c g g t c c c a t g t a a c c g t c a t g a g g g t a a c t g c a g t t a c c g c

8580 CMV IE94 promoter g t a a a t g g c c c g c g a t g g c t g c c a a g t a c a t c c c c a t t g a c g t c a a t g g g g a g g g g c a a t c a t t t a c c g g g c g c t a c c g a c g g t t c a t g t a g g g g t a a c t g c a g t t a c c c c t c c c c g t t a

8640 CMV IE94 promoter g a c g c a a a t g g g c g t t c c a t t g a c g t a a a t g g g c g g t a g g c g t g c c t a a t g g g a g g t c t a c t g c g t t t a c c c g c a a g g t a a c t g c a t t t a c c c g c c a t c c g c a c g g a t t a c c c t c c a g a t

3ʹ 5ʹ 8663 CMV IE94 promoter t a t a a g c a a t g c t c g t t t a g g g a a t a t t c g t t a c g a g c a a a t c c c t

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.