Addgene pX601-mCherry - Sequence Analyzer Skip to main content
Addgene

Sequence Analyzer: pX601-mCherry


Map View

XbaI (137) pBR322ori-F (7962 .. 7981) SacII (7164) BpmI (7160) FspI (7006) PvuI (6860) Amp-R (6660 .. 6679) pBRforEco (6251 .. 6269) pGEX 3' (6191 .. 6213) pRS-marker (6072 .. 6091) PsiI (5958) F1ori-F (5897 .. 5918) F1ori-R (5687 .. 5706) EagI - NotI (5378) LKO.1 5' (5198 .. 5217) hU6-F (5027 .. 5047) Bsu36I (5025) KpnI (5024) Acc65I (5020) bGH poly(A) signal BGH-rev (4806 .. 4823) EcoRI (4782) SgrAI (4752) BbvCI (4609) BbsI (4506) mCherry-F (4484 .. 4503) mCherry-R (4216 .. 4234) PaeR7I - PspXI - XhoI (143) SpeI (168) SnaBI (509) CMV-F (688 .. 708) AgeI (750) Kozak sequence SV40 NLS BspDI * - ClaI * (895) PpuMI * (1449) HindIII (2165) BspEI (2335) BstAPI (2393) CsiI - SexAI * (2743) nucleoplasmin NLS FseI (3997) BamHI (4026) pX601-mCherry 8122 bp

Sequence View

5ʹ 3ʹ 60 AAV2 ITR (alternate) c c t g c a g g c a g c t g c g c g c t c g c t c g c t c a c t g a g g c c g c c c g g g c g t c g g g c g a c c t t t g g a c g t c c g t c g a c g c g c g a g c g a g c g a g t g a c t c c g g c g g g c c c g c a g c c c g c t g g a a a

120 AAV2 ITR (alternate) g g t c g c c c g g c c t c a g t g a g c g a g c g a g c g c g c a g a g a g g g a g t g g c c a a c t c c a t c a c t c c a g c g g g c c g g a g t c a c t c g c t c g c t c g c g c g t c t c t c c c t c a c c g g t t g a g g t a g t g a

SpeIPaeR7IPspXIXhoIXbaI 180 AAV2 ITR (alternate) CMV enhancer a g g g g t t c c t g c g g c c t c t a g a c t c g a g g c g t t g a 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 t c c c c a a g g a c g c c g g a g a t c t g a g c t c c g c a a c t 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

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

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

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

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

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

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

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

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

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 MGGRRVRWE a a t g t c g t a a c a a c t c c g c c c c a t t g a c g c a a a t g g g c g g t a g g c g t g t a c g g t g g g a g g t t a c a g c a t t g t t g a g g c g g g g t a a c t g c g t t t a c c c g c c a t c c g c a c a t g c c a c c c t c c

AgeI 780 PKKK CMV promoter Kozak sequence SV40 NLS VYISRALWLTTGATMAPKKK *SGTGGHGWLLL t c t a t a t a a g c a g a g c t c t c t g g c t a a c t a c c g g t g c c a c c a t g g c c c c a a a g a a g a a g c a g a t a t a t t c g t c t c g a g a g a c c g a t t g a t g g c c a c g g t g g t a c c g g g g t t t c t t c t t c g

840 130 RKV (in frame with SV40 NLS) GIHGVPAA 5 10 KRNYILGLD SV40 NLS SaCas9 RKVGIHGVPAAKRNYILGLD PLDTDVSDWCGLPVVDQAQV g g a a g g t c g g t a t c c a c g g a g t c c c a g c a g c c a a g c g g a a c t a c a t c c t g g g c c t g g a c a c c t t c c a g c c a t a g g t g c c t c a g g g t c g t c g g t t c g c c t t g a t g t a g g a c c c g g a c c t g t

BspDI*ClaI* 900 15 20 25 30 IGITSVGYGIIDYETRDVID SaCas9 IGITSVGYGIIDYETRDVID DADGAHAVADDVVLCPVHDI t c g g c a t c a c c a g c g t g g g c t a c g g c a t c a t c g a c t a c g a g a c a c g g g a c g t g a t c g a t g a g c c g t a g t g g t c g c a c c c g a t g c c g t a g t a g c t g a t g c t c t g t g c c c t g c a c t a g c t a c

960 35 40 45 50 AGVRLFKEANVENNEGRRSK SaCas9 AGVRLFKEANVENNEGRRSK GAHPQEFLGVHFVVLAPPAL c c g g c g t g c g g c t g t t c a a a g a g g c c a a c g t g g a a a a c a a c g a g g g c a g g c g g a g c a a g a g g c c g c a c g c c g a c a a g t t t c t c c g g t t g c a c c t t t t g t t g c t c c c g t c c g c c t c g t t c t

1020 55 60 65 70 RGARRLKRRRRHRIQRVKKL SaCas9 RGARRLKRRRRHRIQRVKKL SAGSPQLPPPPMSDLSHLLQ g a g g c g c c a g a a g g c t g a a g c g g c g g a g g c g g c a t a g a a t c c a g a g a g t g a a g a a g c t g c c t c c g c g g t c t t c c g a c t t c g c c g c c t c c g c c g t a t c t t a g g t c t c t c a c t t c t t c g a c g

1080 75 80 85 90 LFDYNLLTDHSELSGINPYE SaCas9 LFDYNLLTDHSELSGINPYE QEVVVQQGVVALQAADVGVL t g t t c g a c t a c a a c c t g c t g a c c g a c c a c a g c g a g c t g a g c g g c a t c a a c c c c t a c g a g g a c a a g c t g a t g t t g g a c g a c t g g c t g g t g t c g c t c g a c t c g c c g t a g t t g g g g a t g c t c c

1140 95 100 105 110 ARVKGLSQKLSEEEFSAALL SaCas9 ARVKGLSQKLSEEEFSAALL GSHLAQALLQALFLERGGQQ c c a g a g t g a a g g g c c t g a g c c a g a a g c t g a g c g a g g a a g a g t t c t c t g c c g c c c t g c t g c g g t c t c a c t t c c c g g a c t c g g t c t t c g a c t c g c t c c t t c t c a a g a g a c g g c g g g a c g a c g

1200 115 120 125 130 HLAKRRGVHNVNEVEEDTGN SaCas9 HLAKRRGVHNVNEVEEDTGN VQGLSSAHVVHVLHFLVGAV a c c t g g c c a a g a g a a g a g g c g t g c a c a a c g t g a a c g a g g t g g a a g a g g a c a c c g g c a a c g t g g a c c g g t t c t c t t c t c c g c a c g t g t t g c a c t t g c t c c a c c t t c t c c t g t g g c c g t t g c

1260 135 140 145 150 ELSTKEQISRNSKALEEKYV SaCas9 ELSTKEQISRNSKALEEKYV LQGGFLLDAPVALGQFLFVH a g c t g t c c a c c a a a g a g c a g a t c a g c c g g a a c a g c a a g g c c c t g g a a g a g a a a t a c g t g g t c g a c a g g t g g t t t c t c g t c t a g t c g g c c t t g t c g t t c c g g g a c c t t c t c t t t a t g c a c c

1320 155 160 165 170 AELQLERLKKDGEVRGSINR SaCas9 AELQLERLKKDGEVRGSINR GFQLQFPQLFVAFHPAADVS c c g a a c t g c a g c t g g a a c g g c t g a a g a a a g a c g g c g a a g t g c g g g g c a g c a t c a a c a g a t g g c t t g a c g t c g a c c t t g c c g a c t t c t t t c t g c c g c t t c a c g c c c c g t c g t a g t t g t c t a

1380 175 180 185 190 FKTSDYVKEAKQLLKVQKAY SaCas9 FKTSDYVKEAKQLLKVQKAY ELGAVVHFFGFLQQLHLLGV t c a a g a c c a g c g a c t a c g t g a a a g a a g c c a a a c a g c t g c t g a a g g t g c a g a a g g c c t a c c a g t t c t g g t c g c t g a t g c a c t t t c t t c g g t t t g t c g a c g a c t t c c a c g t c t t c c g g a t g g

1440 195 200 205 210 HQLDQSFIDTYIDLLETRRT SaCas9 HQLDQSFIDTYIDLLETRRT VLQVLAEDVGVDVQQFGPPG a c c a g c t g g a c c a g a g c t t c a t c g a c a c c t a c a t c g a c c t g c t g g a a a c c c g g c g g a c c t t g g t c g a c c t g g t c t c g a a g t a g c t g t g g a t g t a g c t g g a c g a c c t t t g g g c c g c c t g g a

PpuMI* 1500 215 220 225 230 YYEGPGEGSPFGWKDIKEWY SaCas9 YYEGPGEGSPFGWKDIKEWY VILSRALAAGEAPLVDFFPV a c t a t g a g g g a c c t g g c g a g g g c a g c c c c t t c g g c t g g a a g g a c a t c a a a g a a t g g t a c g t g a t a c t c c c t g g a c c g c t c c c g t c g g g g a a g c c g a c c t t c c t g t a g t t t c t t a c c a t g c

1560 235 240 245 250 EMLMGHCTYFPEELRSVKYA SaCas9 EMLMGHCTYFPEELRSVKYA LHQHAVAGVEGLFQPAHLVG a g a t g c t g a t g g g c c a c t g c a c c t a c t t c c c c g a g g a a c t g c g g a g c g t g a a g t a c g c c t t c t a c g a c t a c c c g g t g a c g t g g a t g a a g g g g c t c c t t g a c g c c t c g c a c t t c a t g c g g a

1620 255 260 265 270 YNADLYNALNDLNNLVITRD SaCas9 YNADLYNALNDLNNLVITRD VVGVQVVGQVVQVIEHDGPV a c a a c g c c g a c c t g t a c a a c g c c c t g a a c g a c c t g a a c a a t c t c g t g a t c a c c a g g g a c g t g t t g c g g c t g g a c a t g t t g c g g g a c t t g c t g g a c t t g t t a g a g c a c t a g t g g t c c c t g c

1680 275 280 285 290 ENEKLEYYEKFQIIENVFKQ SaCas9 ENEKLEYYEKFQIIENVFKQ LVLLQFIVLLELDDLVHELL a g a a c g a g a a g c t g g a a t a t t a c g a g a a g t t c c a g a t c a t c g a g a a c g t g t t c a a g c a g a t c t t g c t c t t c g a c c t t a t a a t g c t c t t c a a g g t c t a g t a g c t c t t g c a c a a g t t c g t c t

1740 295 300 305 310 KKKPTLKQIAKEILVNEEDI SaCas9 KKKPTLKQIAKEILVNEEDI LLLGGQLLDGFFDEHVFLIN a g a a g a a g c c c a c c c t g a a g c a g a t c g c c a a a g a a a t c c t c g t g a a c g a a g a g g a t a t t a t c t t c t t c g g g t g g g a c t t c g t c t a g c g g t t t c t t t a g g a g c a c t t g c t t c t c c t a t a a t

1800 315 320 325 330 KGYRVTSTGKPEFTNLKVYH SaCas9 KGYRVTSTGKPEFTNLKVYH LAVSHGAGALGLEGVQLHVV a g g g c t a c a g a g t g a c c a g c a c c g g c a a g c c c g a g t t c a c c a a c c t g a a g g t g t a c c a c g t c c c g a t g t c t c a c t g g t c g t g g c c g t t c g g g c t c a a g t g g t t g g a c t t c c a c a t g g t g c

1860 335 340 345 350 DIKDITARKEIIENAELLDQ SaCas9 DIKDITARKEIIENAELLDQ VDLVNGGPFLNNLVGLQQIL a c a t c a a g g a c a t t a c c g c c c g g a a a g a g a t t a t t g a g a a c g c c g a g c t g c t g g a t c a g a t g t a g t t c c t g t a a t g g c g g g c c t t t c t c t a a t a a c t c t t g c g g c t c g a c g a c c t a g t c t

1920 355 360 365 370 IAKILTIYQSSEDIQEELTN SaCas9 IAKILTIYQSSEDIQEELTN NGLDQGDVLAALVDLFFQGI t t g c c a a g a t c c t g a c c a t c t a c c a g a g c a g c g a g g a c a t c c a g g a a g a a c t g a c c a a t c a a c g g t t c t a g g a c t g g t a g a t g g t c t c g t c g c t c c t g t a g g t c c t t c t t g a c t g g t t a g

1980 375 380 385 390 LNSELTQEEIEQISNLKGYT SaCas9 LNSELTQEEIEQISNLKGYT QVGLQGLFLDLLDRIQLAIG *VP t g a a c t c c g a g c t g a c c c a g g a a g a g a t c g a g c a g a t c t c t a a t c t g a a g g g c t a t a c c g a c t t g a g g c t c g a c t g g g t c c t t c t c t a g c t c g t c t a g a g a t t a g a c t t c c c g a t a t g g c

2040 395 400 405 410 GTHNLSLKAINLILDELWHT SaCas9 GTHNLSLKAINLILDELWHT AGVVQAQLGDVQDQVLQPVG VWLRLRFAMLRIRSSSHCVL g c a c c c a c a a c c t g a g c c t g a a g g c c a t c a a c c t g a t c c t g g a c g a g c t g t g g c a c a c c a c g t g g g t g t t g g a c t c g g a c t t c c g g t a g t t g g a c t a g g a c c t g c t c g a c a c c g t g t g g t

2100 415 420 425 430 NDNQIAIFNRLKLVPKKVDL SaCas9 NDNQIAIFNRLKLVPKKVDL VVVLDSDEVPQLQHGLLHVQ SLWIAIKLRSFSTGLFTSRD a c g a c a a c c a g a t c g c t a t c t t c a a c c g g c t g a a g c t g g t g c c c a a g a a g g t g g a c c t g t t g c t g t t g g t c t a g c g a t a g a a g t t g g c c g a c t t c g a c c a c g g g t t c t t c c a c c t g g a c a

2160 435 440 445 450 SQQKEIPTTLVDDFILSPVV SaCas9 SQQKEIPTTLVDDFILSPVV GLLFLDGGGQHVVEDQAGDH WCFSIGVVRTSSKMRLGTTF c c c a g c a g a a a g a g a t c c c c a c c a c c c t g g t g g a c g a c t t c a t c c t g a g c c c c g t c g t g a g g g t c g t c t t t c t c t a g g g g t g g t g g g a c c a c c t g c t g a a g t a g g a c t c g g g g c a g c a c t

HindIII 2220 455 460 465 470 KRSFIQSIKVINAIIKKYGL SaCas9 KRSFIQSIKVINAIIKKYGL LSAEDLADFHDVGDDLLVAQ LLKMWLMLTILAMMLFYPRG a g a g a a g c t t c a t c c a g a g c a t c a a a g t g a t c a a c g c c a t c a t c a a g a a g t a c g g c c t g c t c t c t t c g a a g t a g g t c t c g t a g t t t c a c t a g t t g c g g t a g t a g t t c t t c a t g c c g g a c g

2280 475 480 485 490 PNDIIIELAREKNSKDAQKM SaCas9 PNDIIIELAREKNSKDAQKM GVVDNDLQGALLVGLVGLFH LSMMISSARSFFELSAWFII c c a a c g a c a t c a t t a t c g a g c t g g c c c g c g a g a a g a a c t c c a a g g a c g c c c a g a a a a t g a g g t t g c t g t a g t a a t a g c t c g a c c g g g c g c t c t t c t t g a g g t t c c t g c g g g t c t t t t a c t

BspEI 2340 495 500 505 510 INEMQKRNRQTNERIEEIIR SaCas9 INEMQKRNRQTNERIEEIIR DVLHLLPVPLGVLPDLFDDP LSICFRFRCVLSRISSIMRV t c a a c g a g a t g c a g a a g c g g a a c c g g c a g a c c a a c g a g c g g a t c g a g g a a a t c a t c c g g a a g t t g c t c t a c g t c t t c g c c t t g g c c g t c t g g t t g c t c g c c t a g c t c c t t t a g t a g g c c t

BstAPI 2400 515 520 525 530 TTGKENAKYLIEKIKLHDMQ SaCas9 TTGKENAKYLIEKIKLHDMQ GGAFLVGLVQDLLDLQVVHL VPLSFALYRISFILSCSMCS c c a c c g g c a a a g a g a a c g c c a a g t a c c t g a t c g a g a a g a t c a a g c t g c a c g a c a t g c a g g g g t g g c c g t t t c t c t t g c g g t t c a t g g a c t a g c t c t t c t a g t t c g a c g t g c t g t a c g t c c

2460 535 540 545 550 EGKCLYSLEAIPLEDLLNNP SaCas9 EGKCLYSLEAIPLEDLLNNP FALAQVAQFGDRQFIQQVVG PLHRYLRSAM a a g g c a a g t g c c t g t a c a g c c t g g a a g c c a t c c c t c t g g a a g a t c t g c t g a a c a a c c c c t t t c c g t t c a c g g a c a t g t c g g a c c t t c g g t a g g g a g a c c t t c t a g a c g a c t t g t t g g g g a

2520 555 560 565 570 FNYEVDHIIPRSVSFDNSFN SaCas9 FNYEVDHIIPRSVSFDNSFN EVILHVVDDGSAHGEVVAEV t c a a c t a t g a g g t g g a c c a c a t c a t c c c c a g a a g c g t g t c c t t c g a c a a c a g c t t c a a c a a g t t g a t a c t c c a c c t g g t g t a g t a g g g g t c t t c g c a c a g g a a g c t g t t g t c g a a g t t g t

2580 575 580 585 590 NKVLVKQEENSKKGNRTPFQ SaCas9 NKVLVKQEENSKKGNRTPFQ VLHEHLLFFVALLAVPGWEL a c a a g g t g c t c g t g a a g c a g g a a g a a a a c a g c a a g a a g g g c a a c c g g a c c c c a t t c c a g t t g t t c c a c g a g c a c t t c g t c c t t c t t t t g t c g t t c t t c c c g t t g g c c t g g g g t a a g g t c a

2640 595 600 605 610 YLSSSDSKISYETFKKHILN SaCas9 YLSSSDSKISYETFKKHILN VQAAAVALDAVFGELLVDQI a c c t g a g c a g c a g c g a c a g c a a g a t c a g c t a c g a a a c c t t c a a g a a g c a c a t c c t g a a t c t g g a c t c g t c g t c g c t g t c g t t c t a g t c g a t g c t t t g g a a g t t c t t c g t g t a g g a c t t a g

2700 615 620 625 630 LAKGKGRISKTKKEYLLEER SaCas9 LAKGKGRISKTKKEYLLEER QGLALASDALGLFLIQQFFP t g g c c a a g g g c a a g g g c a g a a t c a g c a a g a c c a a g a a a g a g t a t c t g c t g g a a g a a c g g g a c c g g t t c c c g t t c c c g t c t t a g t c g t t c t g g t t c t t t c t c a t a g a c g a c c t t c t t g c c c

SexAI*CsiI 2760 635 640 645 650 DINRFSVQKDFINRNLVDTR SaCas9 DINRFSVQKDFINRNLVDTR VDVPEGHLFVEDVPVQHIGS a c a t c a a c a g g t t c t c c g t g c a g a a a g a c t t c a t c a a c c g g a a c c t g g t g g a t a c c a g a t t g t a g t t g t c c a a g a g g c a c g t c t t t c t g a a g t a g t t g g c c t t g g a c c a c c t a t g g t c t a

2820 655 660 665 670 YATRGLMNLLRSYFRVNNLD SaCas9 YATRGLMNLLRSYFRVNNLD VGGSAQHVQQPAVESHVVQV a c g c c a c c a g a g g c c t g a t g a a c c t g c t g c g g a g c t a c t t c a g a g t g a a c a a c c t g g a c g t g c g g t g g t c t c c g g a c t a c t t g g a c g a c g c c t c g a t g a a g t c t c a c t t g t t g g a c c t g c

2880 675 680 685 690 VKVKSINGGFTSFLRRKWKF SaCas9 VKVKSINGGFTSFLRRKWKF HFHLGDIAAEGAKQPPLPLK t g a a a g t g a a g t c c a t c a a t g g c g g c t t c a c c a g c t t t c t g c g g c g g a a g t g g a a g t t t a a c t t t c a c t t c a g g t a g t t a c c g c c g a a g t g g t c g a a a g a c g c c g c c t t c a c c t t c a a a t

2940 695 700 705 710 KKERNKGYKHHAEDALIIAN SaCas9 KKERNKGYKHHAEDALIIAN LFLPVLPVLVVGLVGQDNGV a g a a a g a g c g g a a c a a g g g g t a c a a g c a c c a c g c c g a g g a c g c c c t g a t c a t t g c c a a c g t c t t t c t c g c c t t g t t c c c c a t g t t c g t g g t g c g g c t c c t g c g g g a c t a g t a a c g g t t g c

3000 715 720 725 730 ADFIFKEWKKLDKAKKVMEN SaCas9 ADFIFKEWKKLDKAKKVMEN GIEDEFLPLFQVLGFFHHFV c c g a t t t c a t c t t c a a a g a g t g g a a g a a a c t g g a c a a g g c c a a a a a a g t g a t g g a a a a c c g g c t a a a g t a g a a g t t t c t c a c c t t c t t t g a c c t g t t c c g g t t t t t t c a c t a c c t t t t g g

3060 735 740 745 750 QMFEEKQAESMPEIETEQEY SaCas9 QMFEEKQAESMPEIETEQEY LHELFLLGLAHGLDFGLLLV a g a t g t t c g a g g a a a a g c a g g c c g a g a g c a t g c c c g a g a t c g a a a c c g a g c a g g a g t a c a t c t a c a a g c t c c t t t t c g t c c g g c t c t c g t a c g g g c t c t a g c t t t g g c t c g t c c t c a t g t

3120 755 760 765 770 KEIFITPHQIKHIKDFKDYK SaCas9 KEIFITPHQIKHIKDFKDYK FLDEDGGVLDLVNLVELVVL a a g a g a t c t t c a t c a c c c c c c a c c a g a t c a a g c a c a t t a a g g a c t t c a a g g a c t a c a a g t t t c t c t a g a a g t a g t g g g g g g t g g t c t a g t t c g t g t a a t t c c t g a a g t t c c t g a t g t t c a

3180 775 780 785 790 YSHRVDKKPNRELINDTLYS SaCas9 YSHRVDKKPNRELINDTLYS VAVPHVLLRISLQNVVGQVG a c a g c c a c c g g g t g g a c a a g a a g c c t a a t a g a g a g c t g a t t a a c g a c a c c c t g t a c t c c a t g t c g g t g g c c c a c c t g t t c t t c g g a t t a t c t c t c g a c t a a t t g c t g t g g g a c a t g a g g t

3240 795 800 805 810 TRKDDKGNTLIVNNLNGLYD SaCas9 TRKDDKGNTLIVNNLNGLYD GPLVVLAVGQDHVIQVAQVV c c c g g a a g g a c g a c a a g g g c a a c a c c c t g a t c g t g a a c a a t c t g a a c g g c c t g t a c g a c a g g g c c t t c c t g c t g t t c c c g t t g t g g g a c t a g c a c t t g t t a g a c t t g c c g g a c a t g c t g t

3300 815 820 825 830 KDNDKLKKLINKSPEKLLMY SaCas9 KDNDKLKKLINKSPEKLLMY LVIVLQFLQDVLAGFLQQHV a g g a c a a t g a c a a g c t g a a a a a g c t g a t c a a c a a g a g c c c c g a a a a g c t g c t g a t g t a c c t c c t g t t a c t g t t c g a c t t t t t c g a c t a g t t g t t c t c g g g g c t t t t c g a c g a c t a c a t g g

3360 835 840 845 850 HHDPQTYQKLKLIMEQYGDE SaCas9 HHDPQTYQKLKLIMEQYGDE VVVGLGVLFQLQNHFLVAVL a c c a c g a c c c c c a g a c c t a c c a g a a a c t g a a g c t g a t t a t g g a a c a g t a c g g c g a c g a g a t g g t g c t g g g g g t c t g g a t g g t c t t t g a c t t c g a c t a a t a c c t t g t c a t g c c g c t g c t c t

3420 855 860 865 870 KNPLYKYYEETGNYLTKYSK SaCas9 KNPLYKYYEETGNYLTKYSK LIGQVLVVLFGPVVQGLVGF a g a a t c c c c t g t a c a a g t a c t a c g a g g a a a c c g g g a a c t a c c t g a c c a a g t a c t c c a a a a t c t t a g g g g a c a t g t t c a t g a t g c t c c t t t g g c c c t t g a t g g a c t g g t t c a t g a g g t t t t

3480 875 880 885 890 KDNGPVIKKIKYYGNKLNAH SaCas9 KDNGPVIKKIKYYGNKLNAH LVVAGHDLLNLIVAVFQVGM a g g a c a a c g g c c c c g t g a t c a a g a a g a t t a a g t a t t a c g g c a a c a a a c t g a a c g c c c a t c t c c t g t t g c c g g g g c a c t a g t t c t t c t a a t t c a t a a t g c c g t t g t t t g a c t t g c g g g t a g

3540 895 900 905 910 LDITDDYPNSRNKVVKLSLK SaCas9 LDITDDYPNSRNKVVKLSLK t g g a c a t c a c c g a c g a c t a c c c c a a c a g c a g a a a c a a g g t c g t g a a g c t g t c c c t g a a g c a c c t g t a g t g g c t g c t g a t g g g g t t g t c g t c t t t g t t c c a g c a c t t c g a c a g g g a c t t c g

3600 915 920 925 930 PYRFDVYLDNGVYKFVTVKN SaCas9 PYRFDVYLDNGVYKFVTVKN c c t a c a g a t t c g a c g t g t a c c t g g a c a a t g g c g t g t a c a a g t t c g t g a c c g t g a a g a a t c g g a t g t c t a a g c t g c a c a t g g a c c t g t t a c c g c a c a t g t t c a a g c a c t g g c a c t t c t t a g

3660 935 940 945 950 LDVIKKENYYEVNSKCYEEA SaCas9 LDVIKKENYYEVNSKCYEEA t g g a t g t g a t c a a a a a a g a a a a c t a c t a c g a a g t g a a t a g c a a g t g c t a t g a g g a a g c t a a c c t a c a c t a g t t t t t t c t t t t g a t g a t g c t t c a c t t a t c g t t c a c g a t a c t c c t t c g a t

3720 955 960 965 970 KKLKKISNQAEFIASFYNND SaCas9 KKLKKISNQAEFIASFYNND a g a a g c t g a a g a a g a t c a g c a a c c a g g c c g a g t t t a t c g c c t c c t t c t a c a a c a a c g a t c t c t t c g a c t t c t t c t a g t c g t t g g t c c g g c t c a a a t a g c g g a g g a a g a t g t t g t t g c t a g

3780 975 980 985 990 LIKINGELYRVIGVNNDLLN SaCas9 LIKINGELYRVIGVNNDLLN t g a t c a a g a t c a a c g g c g a g c t g t a t a g a g t g a t c g g c g t g a a c a a c g a c c t g c t g a a c c a c t a g t t c t a g t t g c c g c t c g a c a t a t c t c a c t a g c c g c a c t t g t t g c t g g a c g a c t t g g

3840 995 1000 1005 1010 RIEVNMIDITYREYLENMND SaCas9 RIEVNMIDITYREYLENMND g g a t c g a a g t g a a c a t g a t c g a c a t c a c c t a c c g c g a g t a c c t g g a a a a c a t g a a c g a c a c c t a g c t t c a c t t g t a c t a g c t g t a g t g g a t g g c g c t c a t g g a c c t t t t g t a c t t g c t g t

3900 1015 1020 1025 1030 KRPPRIIKTIASKTQSIKKY SaCas9 KRPPRIIKTIASKTQSIKKY a g a g g c c c c c c a g g a t c a t t a a g a c a a t c g c c t c c a a g a c c c a g a g c a t t a a g a a g t a c a t c t c c g g g g g g t c c t a g t a a t t c t g t t a g c g g a g g t t c t g g g t c t c g t a a t t c t t c a t g t

3960 1035 1040 1045 1050 STDILGNLYEVKSKKHPQII SaCas9 STDILGNLYEVKSKKHPQII g c a c a g a c a t t c t g g g c a a c c t g t a t g a a g t g a a a t c t a a g a a g c a c c c t c a g a t c a t c a c g t g t c t g t a a g a c c c g t t g g a c a t a c t t c a c t t t a g a t t c t t c g t g g g a g t c t a g t a g t

FseI 4020 KKG 1 5 10 15 KRPAATKKAGQAKKKK E SaCas9 nucleoplasmin NLS T2A KKGKRPAATKKAGQAKKKKE a a a a g g g c a a a a g g c c g g c g g c c a c g a a a a a g g c c g g c c a g g c a a a a a a g a a a a a g g a g g t t t t c c c g t t t t c c g g c c g c c g g t g c t t t t t c c g g c c g g t c c g t t t t t t c t t t t t c c t c c

BamHI 4080 140 145 150 155 GRGSLLTCGDVEENPGP 1 MVS T2A mCherry GRGSLLTCGDVEENPGPMVS g c a g a g g a t c c c t g c t a a c a t g t g g t g a c g t c g a g g a g a a t c c t g g c c c a a t g g t g a g c a c g t c t c c t a g g g a c g a t t g t a c a c c a c t g c a g c t c c t c t t a g g a c c g g g t t a c c a c t c g t

4140 5 10 15 20 KGEEDNMAIIKEFMRFKVHM mCherry KGEEDNMAIIKEFMRFKVHM a g g g c g a g g a g g a t a a c a t g g c c a t c a t c a a g g a g t t c a t g c g c t t c a a g g t g c a c a t g g t c c c g c t c c t c c t a t t g t a c c g g t a g t a g t t c c t c a a g t a c g c g a a g t t c c a c g t g t a c c

4200 25 30 35 40 EGSVNGHEFEIEGEGEGRPY mCherry EGSVNGHEFEIEGEGEGRPY a g g g c t c c g t g a a c g g c c a c g a g t t c g a g a t c g a g g g c g a g g g c g a g g g c c g c c c c t a c g t c c c g a g g c a c t t g c c g g t g c t c a a g c t c t a g c t c c c g c t c c c g c t c c c g g c g g g g a t g c

4260 45 50 55 60 EGTQTAKLKVTKGGPLPFAW mCherry G G T T C G A C T T C C A C T G G T T mCherry-R EGTQTAKLKVTKGGPLPFAW a g g g c a c c c a g a c c g c c a a g c t g a a g g t g a c c a a g g g t g g c c c c c t g c c c t t c g c c t g g g t c c c g t g g g t c t g g c g g t t c g a c t t c c a c t g g t t c c c a c c g g g g g a c g g g a a g c g g a c c c

4320 65 70 75 80 DILSPQFMYGSKAYVKHPAD mCherry DILSPQFMYGSKAYVKHPAD a c a t c c t g t c c c c t c a g t t c a t g t a c g g c t c c a a g g c c t a c g t g a a g c a c c c c g c c g a c a t g t a g g a c a g g g g a g t c a a g t a c a t g c c g a g g t t c c g g a t g c a c t t c g t g g g g c g g c t g t

4380 85 90 95 100 IPDYLKLSFPEGFKWERVMN mCherry IPDYLKLSFPEGFKWERVMN t c c c c g a c t a c t t g a a g c t g t c c t t c c c c g a g g g c t t c a a g t g g g a g c g c g t g a t g a a c t a g g g g c t g a t g a a c t t c g a c a g g a a g g g g c t c c c g a a g t t c a c c c t c g c g c a c t a c t t g a

4440 105 110 115 120 FEDGGVVTVTQDSSLQDGEF mCherry FEDGGVVTVTQDSSLQDGEF t c g a g g a c g g c g g c g t g g t g a c c g t g a c c c a g g a c t c c t c c c t g c a g g a c g g c g a g t t c a a g c t c c t g c c g c c g c a c c a c t g g c a c t g g g t c c t g a g g a g g g a c g t c c t g c c g c t c a a g t

4500 125 130 135 140 IYKVKLRGTNFPSDGPVMQK mCherry C C C C G T A A T G C A G A A G A mCherry-F IYKVKLRGTNFPSDGPVMQK t c t a c a a g g t g a a g c t g c g c g g c a c c a a c t t c c c c t c c g a c g g c c c c g t a a t g c a g a a g a a g a t g t t c c a c t t c g a c g c g c c g t g g t t g a a g g g g a g g c t g c c g g g g c a t t a c g t c t t c t

BbsI 4560 145 150 155 160 KTMGWEASSERMYPEDGALK mCherry A G A mCherry-F KTMGWEASSERMYPEDGALK a g a c c a t g g g c t g g g a g g c c t c c t c c g a g c g g a t g t a c c c c g a g g a c g g c g c c c t g a a g g t c t g g t a c c c g a c c c t c c g g a g g a g g c t c g c c t a c a t g g g g c t c c t g c c g c g g g a c t t c c

BbvCI 4620 165 170 175 180 GEIKQRLKLKDGGHYDAEVK mCherry GEIKQRLKLKDGGHYDAEVK g c g a g a t c a a g c a g a g g c t g a a g c t g a a g g a c g g c g g c c a c t a c g a c g c t g a g g t c a a g a c g c t c t a g t t c g t c t c c g a c t t c g a c t t c c t g c c g c c g g t g a t g c t g c g a c t c c a g t t c t

4680 185 190 195 200 TTYKAKKPVQLPGAYNVNIK mCherry TTYKAKKPVQLPGAYNVNIK c c a c c t a c a a g g c c a a g a a g c c c g t g c a g c t g c c c g g c g c c t a c a a c g t c a a c a t c a a g t g g t g g a t g t t c c g g t t c t t c g g g c a c g t c g a c g g g c c g c g g a t g t t g c a g t t g t a g t t c a

4740 205 210 215 220 LDITSHNEDYTIVEQYERAE mCherry LDITSHNEDYTIVEQYERAE t g g a c a t c a c c t c c c a c a a c g a g g a c t a c a c c a t c g t g g a a c a g t a c g a a c g c g c c g a g g a c c t g t a g t g g a g g g t g t t g c t c c t g a t g t g g t a g c a c c t t g t c a t g c t t g c g c g g c t c c

EcoRISgrAI 4800 225 230 235 GRHSTGGMDELYK* mCherry GRHSTGGMDELYK* g c c g c c a c t c c a c c g g c g g c a t g g a c g a g c t g t a c a a g t a a g a a t t c c t a g a g c t c g c t g c g g c g g t g a g g t g g c c g c c g t a c c t g c t c g a c a t g t t c a t t c t t a a g g a t c t c g a g c g a c

4860 bGH poly(A) signal G G A G C T G A C A C G G A A G A T BGH-rev a t c a g c c t c g a c t g t g c c t t c t a g t t g c c a g c c a t c t g t t g t t t g c c c c t c c c c c g t g c c t a g t c g g a g c t g a c a c g g a a g a t c a a c g g t c g g t a g a c a a c a a a c g g g g a g g g g g c a c g g

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

4980 bGH poly(A) signal a t c g c a t t g t c t g a g t a g g t g t c a t t c t a t t c t g g g g g g t g g g g t g g g g c a g g a c a g c a a t a g c g t a a c a g a c t c a t c c a c a g t a a g a t a a g a c c c c c c a c c c c a c c c c g t c c t g t c g t t

Acc65IKpnIBsu36I 5040 bGH poly(A) signal U6 promoter G A G G G C C T A T T T C C hU6-F g g g g g a g g a t t g g g a a g a g a a t a g c a g g c a t g c t g g g g a g g t a c c t g a g g g c c t a t t t c c c c c c c t c c t a a c c c t t c t c t t a t c g t c c g t a c g a c c c c t c c a t g g a c t c c c g g a t a a a g g

5100 U6 promoter C A T G A T T hU6-F c a t g a t t c c t t c a t a t t t g c a t a t a c g a t a c a a g g c t g t t a g a g a g a t a a t t g g a a t t a a g t a c t a a g g a a g t a t a a a c g t a t a t g c t a t g t t c c g a c a a t c t c t c t a t t a a c c t t a a t t

5160 U6 promoter t t t g a c t g t a a a c a c a a a g a t a t t a g t a c a a a a t a c g t g a c g t a g a a a g t a a t a a t t t c t a a a c t g a c a t t t g t g t t t c t a t a a t c a t g t t t t a t g c a c t g c a t c t t t c a t t a t t a a a g a

5220 U6 promoter G A C T A T C A T A T G C T T A C C G T LKO.1 5' t g g g t a g t t t g c a g t t t t a a a a t t a t g t t t t a a a a t g g a c t a t c a t a t g c t t a c c g t a a c a c c c a t c a a a c g t c a a a a t t t t a a t a c a a a a t t t t a c c t g a t a g t a t a c g a a t g g c a t t g

5280 U6 promoter t t g a a a g t a t t t c g a t t t c t t g g c t t t a t a t a t c t t g t g g a a a g g a c g a a a c a c c g g a g a a a c t t t c a t a a a g c t a a a g a a c c g a a a t a t a t a g a a c a c c t t t c c t g c t t t g t g g c c t c t

5340 Sa gRNA scaffold c c a c g g c a g g t c t c a g t t t t a g t a c t c t g g a a a c a g a a t c t a c t a a a a c a a g g c a a a a t g g g t g c c g t c c a g a g t c a a a a t c a t g a g a c c t t t g t c t t a g a t g a t t t t g t t c c g t t t t a c

EagINotI 5400 Sa gRNA scaffold AAV2 ITR AAV2 ITR c c g t g t t t a t c t c g t c a a c t t g t t g g c g a g a t t t t t g c g g c c g c a g g a a c c c c t a g t g a t g g c a c a a a t a g a g c a g t t g a a c a a c c g c t c t a a a a a c g c c g g c g t c c t t g g g g a t c a c t a

5460 AAV2 ITR AAV2 ITR g g a g t t g g c c a c t c c c t c t c t g c g c g c t c g c t c g c t c a c t g a g g c c g g g c g a c c a a a g g t c c t c a a c c g g t g a g g g a g a g a c g c g c g a g c g a g c g a g t g a c t c c g g c c c g c t g g t t t c c a

5520 AAV2 ITR AAV2 ITR c g c c c g a c g c c c g g g c t t t g c c c g g g c g g c c t c a g t g a g c g a g c g a g c g c g c a g c t g c c t g c g g g c t g c g g g c c c g a a a c g g g c c c g c c g g a g t c a c t c g c t c g c t c g c g c g t c g a c g g a

5580 AAV2 ITR g c a g g g g c g c c t g a t g c g g t a t t t t c t c c t t a c g c a t c t g t g c g g t a t t t c a c a c c g c a t c g t c c c c g c g g a c t a c g c c a t a a a a g a g g a a t g c g t a g a c a c g c c a t a a a g t g t g g c g t a

5640 f1 ori a c g t c a a a g c a a c c a t a g t a c g c g c c c t g t a g c g g c g c a t t a a g c g c g g c g g g t g t g g t g t g c a g t t t c g t t g g t a t c a t g c g c g g g a c a t c g c c g c g t a a t t c g c g c c g c c c a c a c c a c

5700 f1 ori G A G G A A A G C G A A A G F1ori-R g t t a c g c g c a g c g t g a c c g c t a c a c t t g c c a g c g c c c t a g c g c c c g c t c c t t t c g c t t t c c a a t g c g c g t c g c a c t g g c g a t g t g a a c g g t c g c g g g a t c g c g g g c g a g g a a a g c g a a a g

5760 f1 ori A A G G G A F1ori-R t t c c c t t c c t t t c t c g c c a c g t t c g c c g g c t t t c c c c g t c a a g c t c t a a a t c g g g g g c t c a a g g g a a g g a a a g a g c g g t g c a a g c g g c c g a a a g g g g c a g t t c g a g a t t t a g c c c c c g a g

5820 f1 ori c c t t t a g g g t t c c g a t t t a g t g c t t t a c g g c a c c t c g a c c c c a a a a a a c t t g a t t t g g g t g g a a a t c c c a a g g c t a a a t c a c g a a a t g c c g t g g a g c t g g g g t t t t t t g a a c t a a a c c c a

5880 f1 ori g a t g g t t c a c g t a g t g g g c c a t c g c c c t g a t a g a c g g t t t t t c g c c c t t t g a c g t t g g a g c t a c c a a g t g c a t c a c c c g g t a g c g g g a c t a t c t g c c a a a a a g c g g g a a a c t g c a a c c t c

5940 f1 ori G T G G A C T C T T G T T C C A A A C T G G F1ori-F t c c a c g t t c t t t a a t a g t g g a c t c t t g t t c c a a a c t g g a a c a a c a c t c a a c c c t a t c t c g a g g t g c a a g a a a t t a t c a c c t g a g a a c a a g g t t t g a c c t t g t t g t g a g t t g g g a t a g a g c

PsiI 6000 f1 ori g g c t a t t c t t t t g a t t t a t a a g g g a t t t t g c c g a t t t c g g c c t a t t g g t t a a a a a a t g a g c c g a t a a g a a a a c t a a a t a t t c c c t a a a a c g g c t a a a g c c g g a t a a c c a a t t t t t t a c t c

6060 f1 ori c t g a t t t a a c a a a a a t t t a a c g c g a a t t t t a a c a a a a t a t t a a c g t t t a c a a t t t t a t g g g a c t a a a t t g t t t t t a a a t t g c g c t t a a a a t t g t t t t a t a a t t g c a a a t g t t a a a a t a c c

6120 A T G T T A G A C G A G A C T A C G G C pRS-marker t g c a c t c t c a g t a c a a t c t g c t c t g a t g c c g c a t a g t t a a g c c a g c c c c g a c a c c c g c c a a c g t g a g a g t c a t g t t a g a c g a g a c t a c g g c g t a t c a a t t c g g t c g g g g c t g t g g g c g g t

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

6240 C C G G G A G C T G C A T G T G T C A G A G G pGEX 3' g t g a c c g t c t c c g g g a g c t g c a t g t g t c a g a g g t t t t c a c c g t c a t c a c c g a a a c g c g c g c a c t g g c a g a g g c c c t c g a c g t a c a c a g t c t c c a a a a g t g g c a g t a g t g g c t t t g c g c g c

6300 C G G A G C A C T A T G C G G A T A A pBRforEco a g a c g a a a g g g c c t c g t g a t a c g c c t a t t t t t a t a g g t t a a t g t c a t g a t a a t a a t g g t t t c t g c t t t c c c g g a g c a c t a t g c g g a t a a a a a t a t c c a a t t a c a g t a c t a t t a t t a c c a a

6360 AmpR promoter t c t t a g a c g t c a g g t g g c a c t t t t c g g g g a a a t g t g c g c g g a a c c c c t a t t t g t t t a t t t a g a a t c t g c a g t c c a c c g t g a a a a g c c c c t t t a c a c g c g c c t t g g g g a t a a a c a a a t a a a

6420 AmpR promoter t t c t a a a t a c a t t c a a a t a t g t a t c c g c t c a t g a g a c a a t a a c c c t g a t a a a t g c t t c a a a a g a t t t a t g t a a g t t t a t a c a t a g g c g a g t a c t c t g t t a t t g g g a c t a t t t a c g a a g t t

6480 1 5 10 signal sequence MSIQHFRVALIPF AmpR promoter AmpR MSIQHFRVALIPF t a a t a t t g a a a a a g g a a g a g t a t g a g t a t t c a a c a t t t c c g t g t c g c c c t t a t t c c c t t t a t t a t a a c t t t t t c c t t c t c a t a c t c a t a a g t t g t a a a g g c a c a g c g g g a a t a a g g g a a a

6540 15 20 signal sequence FAAFCLPVFA 25 30 HPETLVKVKD AmpR FAAFCLPVFAHPETLVKVKD t t t g c g g c a t t t t g c c t t c c t g t t t t t g c t c a c c c a g a a a c g c t g g t g a a a g t a a a a g a t a a a c g c c g t a a a a c g g a a g g a c a a a a a c g a g t g g g t c t t t g c g a c c a c t t t c a t t t t c t a

6600 35 40 45 50 AEDQLGARVGYIELDLNSGK AmpR AEDQLGARVGYIELDLNSGK g c t g a a g a t c a g t t g g g t g c a c g a g t g g g t t a c a t c g a a c t g g a t c t c a a c a g c g g t a a g c g a c t t c t a g t c a a c c c a c g t g c t c a c c c a a t g t a g c t t g a c c t a g a g t t g t c g c c a t t c

6660 55 60 65 70 ILESFRPEERFPMMSTFKVL AmpR C Amp-R ILESFRPEERFPMMSTFKVL a t c c t t g a g a g t t t t c g c c c c g a a g a a c g t t t t c c a a t g a t g a g c a c t t t t a a a g t t c t g t a g g a a c t c t c a a a a g c g g g g c t t c t t g c a a a a g g t t a c t a c t c g t g a a a a t t t c a a g a c

6720 75 80 85 90 LCGAVLSRIDAGQEQLGRRI AmpR G A T A C A C C G C G C C A T A A T A Amp-R LCGAVLSRIDAGQEQLGRRI c t a t g t g g c g c g g t a t t a t c c c g t a t t g a c g c c g g g c a a g a g c a a c t c g g t c g c c g c a t a g a t a c a c c g c g c c a t a a t a g g g c a t a a c t g c g g c c c g t t c t c g t t g a g c c a g c g g c g t a t

6780 95 100 105 110 HYSQNDLVEYSPVTEKHLTD AmpR HYSQNDLVEYSPVTEKHLTD c a c t a t t c t c a g a a t g a c t t g g t t g a g t a c t c a c c a g t c a c a g a a a a g c a t c t t a c g g a t g t g a t a a g a g t c t t a c t g a a c c a a c t c a t g a g t g g t c a g t g t c t t t t c g t a g a a t g c c t a

6840 115 120 125 130 GMTVRELCSAAITMSDNTAA AmpR GMTVRELCSAAITMSDNTAA g g c a t g a c a g t a a g a g a a t t a t g c a g t g c t g c c a t a a c c a t g a g t g a t a a c a c t g c g g c c c c g t a c t g t c a t t c t c t t a a t a c g t c a c g a c g g t a t t g g t a c t c a c t a t t g t g a c g c c g g

PvuI 6900 135 140 145 150 NLLLTTIGGPKELTAFLHNM AmpR NLLLTTIGGPKELTAFLHNM a a c t t a c t t c t g a c a a c g a t c g g a g g a c c g a a g g a g c t a a c c g c t t t t t t g c a c a a c a t g t t g a a t g a a g a c t g t t g c t a g c c t c c t g g c t t c c t c g a t t g g c g a a a a a a c g t g t t g t a c

6960 155 160 165 170 GDHVTRLDRWEPELNEAIPN AmpR GDHVTRLDRWEPELNEAIPN g g g g a t c a t g t a a c t c g c c t t g a t c g t t g g g a a c c g g a g c t g a a t g a a g c c a t a c c a a a c c c c c t a g t a c a t t g a g c g g a a c t a g c a a c c c t t g g c c t c g a c t t a c t t c g g t a t g g t t t g

FspI 7020 175 180 185 190 DERDTTMPVAMATTLRKLLT AmpR DERDTTMPVAMATTLRKLLT g a c g a g c g t g a c a c c a c g a t g c c t g t a g c a a t g g c a a c a a c g t t g c g c a a a c t a t t a a c t c t g c t c g c a c t g t g g t g c t a c g g a c a t c g t t a c c g t t g t t g c a a c g c g t t t g a t a a t t g a

7080 195 200 205 210 GELLTLASRQQLIDWMEADK AmpR GELLTLASRQQLIDWMEADK g g c g a a c t a c t t a c t c t a g c t t c c c g g c a a c a a t t a a t a g a c t g g a t g g a g g c g g a t a a a c c g c t t g a t g a a t g a g a t c g a a g g g c c g t t g t t a a t t a t c t g a c c t a c c t c c g c c t a t t t

7140 215 220 225 230 VAGPLLRSALPAGWFIADKS AmpR VAGPLLRSALPAGWFIADKS g t t g c a g g a c c a c t t c t g c g c t c g g c c c t t c c g g c t g g c t g g t t t a t t g c t g a t a a a t c t c a a c g t c c t g g t g a a g a c g c g a g c c g g g a a g g c c g a c c g a c c a a a t a a c g a c t a t t t a g a

BpmISacII 7200 235 240 245 250 GAGERGSRGIIAALGPDGKP AmpR GAGERGSRGIIAALGPDGKP g g a g c c g g t g a g c g t g g a a g c c g c g g t a t c a t t g c a g c a c t g g g g c c a g a t g g t a a g c c c c c t c g g c c a c t c g c a c c t t c g g c g c c a t a g t a a c g t c g t g a c c c c g g t c t a c c a t t c g g g

7260 255 260 265 270 SRIVVIYTTGSQATMDERNR AmpR SRIVVIYTTGSQATMDERNR t c c c g t a t c g t a g t t a t c t a c a c g a c g g g g a g t c a g g c a a c t a t g g a t g a a c g a a a t a g a a g g g c a t a g c a t c a a t a g a t g t g c t g c c c c t c a g t c c g t t g a t a c c t a c t t g c t t t a t c t

7320 275 280 285 QIAEIGASLIKHW* AmpR QIAEIGASLIKHW* c a g a t c g c t g a g a t a g g t g c c t c a c t g a t t a a g c a t t g g t a a c t g t c a g a c c a a g t t t a c g t c t a g c g a c t c t a t c c a c g g a g t g a c t a a t t c g t a a c c a t t g a c a g t c t g g t t c a a a t g

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

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

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

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

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

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

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

7800 ori g g g t t g g a c t c a a g a c g a t a g t t a c c g g a t a a g g c g c a g c g g t c g g g c t g a a c g g g g g g t c c c a a c c t g a g t t c t g c t a t c a a t g g c c t a t t c c g c g t c g c c a g c c c g a c t t g c c c c c c a

7860 ori t c g t g c a c a c a g c c c a g c t t g g a g c g a a c g a c c t a c a c c g a a c t g a g a t a c c t a c a g c g t a g c a c g t g t g t c g g g t c g a a c c t c g c t t g c t g g a t g t g g c t t g a c t c t a t g g a t g t c g c a

7920 ori g a g c t a t g a g a a a g c g c c a c g c t t c c c g a a g g g a g a a a g g c g g a c a g g t a t c c g g t a a g c c t c g a t a c t c t t t c g c g g t g c g a a g g g c t t c c c t c t t t c c g c c t g t c c a t a g g c c a t t c g

7980 ori G G G A A A C G C C T G G T A T C T T pBR322ori-F g g c a g g g t c g g a a c a g g a g a g c g c a c g a g g g a g c t t c c a g g g g g a a a c g c c t g g t a t c t t c c g t c c c a g c c t t g t c c t c t c g c g t g c t c c c t c g a a g g t c c c c c t t t g c g g a c c a t a g a a

8040 ori T pBR322ori-F t a t a g t c c t g t c g g g t t t c g c c a c c t c t g a c t t g a g c g t c g a t t t t t g t g a t g c t c g t c a a t a t c a g g a c a g c c c a a a g c g g t g g a g a c t g a a c t c g c a g c t a a a a a c a c t a c g a g c a g t

8100 ori g g g g g g c g g a g c c t a t g g a a a a a c g c c a g c a a c g c g g c c t t t t t a c g g t t c c t g g c c t t t c c c c c c g c c t c g g a t a c c t t t t t g c g g t c g t t g c g c c g g a a a a a t g c c a a g g a c c g g a a a

3ʹ 5ʹ 8122 t g c t g g c c t t t t g c t c a c a t g t a c g a c c g g a a a a c g a g t g t a c a

Restriction Enzymes

Instructions: By default, all cutters are shown. Filter on number of cut sites or search by enzyme name.

Filter

Features

Primers

BLAST

BLAST (Basic Local Alignment Search Tool) finds regions of similarity between biological sequences. Click on the buttons below to submit a BLAST search to NCBI. The results will appear in a new window. See your recent BLAST results on NCBI's website.

  • Nucleotide-Nucleotide BLAST (BLASTN)

  • Translated Nucleotide-Protein BLAST (BLASTX)

  • Sequence alignment using BLAST (BLAST2)

Sequence Analyzer Guide

Map

Displays a graphical map based on nucleotide sequence data labeled with restriction enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site)

To select a portion of sequence, click one location on the plasmid and then a second location to display the sequence between the two locations.

Sequence

Displays both strands of base paired nucleotide sequences with annotated enzymes, plasmid features, ORFs (theoretical open reading frames) and primers. Hovering over data labels will display additional information (e.g. cut site).

To select a portion of sequence, click one location on the sequence and then a second location to display the sequence between the two locations.

Enzymes

List of restriction enzymes that can cut a given nucleotide sequence. Table lists enzyme name and the sequence location of the cut.

Features

List of common features detected in a given nucleotide sequence. Table lists feature name, location, size, color used to indicate its position on the map, and direction (if relevant).

Primers

List of commonly used primers detected in a given nucleotide sequence. Table lists primer name, sequence, length, binding site location, and direction.

BLAST

Use Basic Local Alignment Search Tool (BLAST) via the NCBI website to determine similarity between a given sequence and nucleotide (BLASTN) or protein (BLASTX) sequence databases. Additionally, align a custom nucleotide sequence against a given sequence using BLAST2.

File Downloads

GenBank

File contains the nucleotide sequence and annotated features in GenBank flat file format. Open the file with a text editor or plasmid mapping software to view the sequence.

SnapGene

File contains the nucleotide sequence and enhanced annotations from SnapGene Server. Open the file with SnapGene software or the free Viewer to view the plasmid map, sequence, and perform additional sequence analysis.