pCOLADuet-1 共表达质粒
合集下载
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1695
2609
494
2003
2267
ቤተ መጻሕፍቲ ባይዱ
3021 1094
3248 2051
2645
473
723 1164
1608 3538
1863
1906
2298
3373
3430 869
2263
2274 Enzymes that do not cut pCOLADuet-1: AarI AfeI AhdI AleI BbvCI BglI BplI BsaAI BsiWI BspEI BspLU11I BssSI BstZ17I DraI DraIII FspAI NspV PciI PmeI PmlI PshAI PsiI PsrI RsrII SapI ScaI SfiI SgrAI SrfI StuI SwaI AloI BsaBI Bst1107I FspI PpiI SacII SnaBI AlwNI BseRI BstBI MscI PpuMI SanDI SpeI
Novagen
Enzyme AatII Acc65I AccI AclI AflII AflIII AgeI ApaI ApaLI AscI AseI AsiSI AvaI AvrII BaeI BamHI BanI BanII BbeI BbsI BceAI BcgI BciVI BclI BfrBI BglII BlpI Bme1580I BmgBI BmrI BmtI BpmI Bpu10I BpuEI BsaHI BsaI BsaWI BsaXI BseYI BsgI BsiEI BsiHKAI BsmAI BsmBI BsmFI BsmI Bsp1286I BspCNI BspHI BspMI BsrBI BsrDI BsrFI BsrGI BssHII BstAPI BstEII BstXI BstYI Bsu36I BtgI BtrI BtsI ClaI DrdI EaeI EagI # Sites 1 1 3 1 1 1 1 1 1 1 6 1 2 1 1 1 4 3 1 2 5 2 4 1 2 1 1 2 1 3 1 2 1 4 4 1 6 2 3 2 8 2 8 2 1 2 4 5 2 1 4 2 4 1 2 1 1 3 7 1 1 1 5 1 1 5 3
Reference 1. Zverev, V.V. and Khmel, I.A. (1985) Plasmid 14, 192-199.
pCOLADuet-1
(3719 bp)
pCOLADuet-1 cloning/expression regions Novagen • ordering 800-526-7319 • technical support 800-207-0144 A Brand of EMD Biosciences, Inc., an Affiliate of Merck KGaA, Darmstadt, Germany
Novagen
pCOLADuet-1 DNA
®
pCOLADuet™-1 Vector
Cat No. 71406-3 pCOLADuet-1 sequence landmarks T7 promoter-1 3703–3719 T7 transcription start-1 1 His•Tag® coding sequence 83–100 Multiple cloning sites-1 (Nco I–Afl II) 68–168 T7 promoter-2 214-230 T7 transcription start-2 231 Multiple cloning sites-2 (Nde I–Avr II) 297–438 S•Tag™ coding sequence 366-410 T7 terminator 462-509 KanR 739–1554 ColA ori 1664-2299 lacI coding sequence 2497-3579
1263
2497
2865
322 322
326
2550
Novagen • ordering 800-526-7319 • technical support 800-207-0144 A Brand of EMD Biosciences, Inc., an Affiliate of Merck KGaA, Darmstadt, Germany
TB408 0304 pCOLADuet™-1 is designed for the coexpression of two target genes from a single plasmid. The vector encodes two multiple cloning sites (MCS) each of which is preceded by a T7 promoter, lac operator, and ribosome binding site (rbs). MCS-1 encodes the six-amino acid His•Tag® sequence for the creation of a N-terminal fusion and MCS2 encodes the 15 amino acid S•Tag™ peptide after the last restriction site for the creation of a C-terminal fusion if desired. Genes inserted into MCS-1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer. Genes inserted into MCS-2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer. The vector has the COLA replicon from ColA(1) and the kanamycin resistance gene. This vector can be transformed into the same cell with plasmids containing compatible origins of replication and drug resistance genes for coexpression of up to 8 target genes.
®
pCOLADuet™-1 Restriction Sites
Locations 346 348 138 411 3564 163 3224 566 3021 3244 125 213 732 3702 337 354 1246 433 365 106 348 2455 122 1471 2589 2739 3078 211 801 162 2904 728 1604 3210 1008 1274 305 451 3021 3248 1908 2426 3066 1658 2903 3392 1102 515 2024 343 1870 645 551 566 2905 655 2556 1939 2690 3179 3379 153 199 1124 2255 122 3248 645 1102 2996 3122 1102 2609 1894 1163 1240 122 1471 443 530 725 1602 124 13 243 2817 3183 324 566 190 125 2813 3548 3042 3178 3301 106 305 2448 3660 517 69 1908 543 1176 1429 637 150 196 150 196 1922 Enzyme EarI EciI Ecl136II Eco57I Eco57MI EcoICRI EcoNI EcoO109I EcoRI EcoRV FalI FseI HaeII HincII HindIII HpaI KasI KpnI MfeI MluI MslI NaeI NarI NcoI NdeI NgoMIV NheI NotI NruI NsiI NspI PacI PflMI PfoI PinAI PspOMI PstI PvuI PvuII SacI SalI SbfI SexAI SfcI SfoI SmaI SmlI SphI Sse8387I SspI StyI TaqII TatI TspGWI Tth111I XbaI XcmI XhoI XmaI XmnI ZraI # Sites 4 2 1 1 3 1 3 1 1 2 1 1 3 2 1 1 1 1 1 1 3 1 1 1 1 1 1 1 1 2 1 1 3 1 1 1 1 2 2 1 1 1 1 4 1 1 6 1 1 2 3 2 1 5 1 1 3 1 1 1 1 Locations 1306 1562 1730 3439 120 2364 2364 2903 120 1209 2107 478 112 319 2080 671 328 2589 2832 139 2722 143 2722 2585 352 311 3224 2858 2888 326 2586 69 298 324 1654 150 1465 1010 1276 1654 429 401 862 3656 566 3017 135 337 2255 2535 2628 122 137 135 2163 29 131 2587 1248 163 354 2431 1654 135 1197 1571 69 433 870 2431 190 1303 1315 637 2304 2839 2857 354 1246 733 344 2332 3607
TB408 0304
3392 3692
921
2482
2541
3613
2585 3021
3304
3176
1708 1681
2740 2772
3367
3303
3649
2252 2586 983
2416 3269 1988 2402
2825 325 2445 1604 3527 337 636
226
3715