pET-16b Vector

pET-16b VectorTB046 2/00The pET-16b vector (Cat. No. 69662-3) carries an N-terminal His•Tag ® sequence followed bya Factor Xa site and three cloning sites. Unique sites are shown on the circle map. Note that thesequence is numbered by the pBR322 convention, so the T7 expression region is reversed on thecircular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymeraseis shown below.pET-16b sequence landmarksT7 promoter 466-482T7 transcription start 465His•Tag coding sequence 360-389Multiple cloning sites(Nde I -BamH I) 319-335T7 terminator 213-259lacI coding sequence 869-1948pBR322 origin 3885bla coding sequence 4646-5503Bsa I(4777)Sca I(5196)Pvu I(5086)Pst I(4961)Eam1105 I(4716) –Aat II(5638)Ssp I(5520)Ap (4646-5503)EcoR I(5709)Cla I(24)Hind III(29)Bpu1102 I(267)BamH I(319)Xho I(324)Nde I(331)Nco I(392)Xba I(431)Bgl II(497)SgrA I(538)Sph I(694)EcoN I(754)Drd II(942)AlwN I(4239) –pET-16b(5711bp)lacI (869-1948)Mlu I(1219)Bcl I(1233)BstE II(1400)Bmg I(1428)Apa I(1430)BssH II(1630)Hpa I(1725)BspLU11 I(3823)ori (3885)BsaX I(1878)Sap I(3707)Bst1107 I(3594)Acc I(3593)BsaA I(3575)Tth111 I(3568)Bpu10 I(2929)Bsm I(2707)Msc I(2794)Eag I(2287)Nru I(2322)BspM I(2402)PshA I(2064)T7 promoter primer #69348-3T7 promoterBgl IIlac operatorXba IrbsAGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGANco IHis•TagNde I Xho I BamH ITATACCATGGGCCATCATCATCATCATCATCATCATCATCACAGCAGCGGCCATATCGAAGGTCGTCATATGCTCGAGGATCCGGCTGCTAACAAAGCCMetGlyHisHisHisHisHisHisHisHisHisHisSerSerGlyHisIleGluGlyArgHisMetLeuGluAspProAlaAlaAsnLysAlaBpu1102 IFactor Xa T7 terminatorCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGArgLysGluAlaGluLeuAlaAlaAlaThrAlaGluGlnEndT7 terminator primer #69337-3pET-16b cloning/expression regionNovagen • ORDERING 800-526-7319 • TECHNICAL SUPPORT 800-207-0144

pET-16b Restriction SitesEnzyme # Sites LocationsAatII 1 5638AccI 1 3593AceIII 7 986 1714 2045 3332 34733775 5015AciI 89AflIII 2 1219 3823AluI 24AlwI 16Alw21I 8 719 1203 2526 2817 36414141 5302 5387Alw44I 4 1199 3637 4137 5383AlwNI 1 4239ApaI 1 1430ApaBI 2 903 2400ApoI 2 1494 5709AvaI 2 324 2773AvaII 9 1771 2147 2235 2484 27872829 3108 4854 5076BamHI 1 319BanI 12BanII 3 603 617 1430BbsI 5 1365 1704 2078 2941 5694BbvI 28BccI 16Bce83I 7 208 2033 2203 3914 42124453 5321BcefI 5 738 1079 1706 2515 4325BcgI 8 1511 1545 2045 2079 34003434 5221 5255BclI 1 1233BfaI 6 257 432 2837 4318 45714906BglI 3 2283 2517 4836BglII 1 497BmgI 1 1428BpmI 6 1057 1546 2180 2734 33504786Bpu10I 1 2929Bpu1102I 1 267BsaI 1 4777BsaAI 1 3575BsaBI 3 496 502 3020BsaHI 8 542 563 677 1176 18592554 5253 5635BsaJI 11BsaWI 7 189 1538 2041 3012 40294176 5007BsaXI 1 1878BsbI 2 3539 5259BscGI 13BsgI 3 1070 1270 2983BsiI 3 3996 5380 5687BsiEI 6 2004 2290 3739 4163 50865235BslI 22BsmI 1 2707BsmAI 7 916 1321 1447 1834 34644777 5553BsmBI 2 1834 3464BsmFI 4 680 2221 2446 3094BsoFI 52Bsp24I 12Bsp1286I 11BspEI 2 189 3012BspGI 3 2407 2484 3349BspLU11I 1 3823BspMI 1 2402BsrI 25BsrBI 3 452 3756 5557BsrDI 4 1266 1632 4777 4951BsrFI 8 160 529 538 905 21172277 2631 4796Enzyme # Sites LocationsBssHII 1 1630Bst1107I 1 3594BstEII 1 1400BstXI 3 1021 1150 1273BstYI 11Cac8I 41CjeI 26CjePI 28ClaI 1 24CviJI 96CviRI 26DdeI 11DpnI 29DraI 3 4582 4601 5293DrdI 2 3516 3931DrdII 1 942DsaI 3 392 656 2795EaeI 7 349 527 659 1893 22872792 5104EagI 1 2287Eam1105I 1 4716EarI 3 837 3707 5511EciI 5 996 2743 3897 4043 4871Eco47III 3 624 2125 3077Eco57I 2 4371 5383EcoNI 1 754EcoO109I 5 240 652 2787 2829 5692EcoRI 1 5709EcoRII 10 129 942 1257 1797 18542406 2789 3849 3970 3983EcoRV 2 187 1669FauI 18FokI 14FspI 3 2706 2804 4938GdiII 6 349 527 659 1893 22875104HaeI 8 947 2268 2340 2397 27943838 3849 4301HaeII 13HaeIII 29HgaI 15HgiEII 2 817 4409HhaI 44Hin4I 5 16 1118 2489 4715 4789HincII 2 1725 5257HindIII 1 29HinfI 14HpaI 1 1725HphI 17MaeII 12MaeIII 18MboII 15MluI 1 1219MmeI 2 4038 4222MnlI 34MscI 1 2794MseI 24MslI 10 1271 1559 1589 2379 28103005 3396 4968 5127 5486MspI 35MspA1I 11MwoI 44NarI 5 542 563 677 1859 2554NciI 14NcoI 1 392NdeI 1 331NgoAIV 4 529 2117 2277 2631NlaIII 31NlaIV 28NruI 1 2322NspI 4 694 3168 3460 3827Pfl1108I 2 2106 4734Enzyme # Sites LocationsPflMI 3 801 2669 2718PleI 7 480 768 855 1651 37174202 4705PshAI 1 2064Psp5II 2 2787 2829Psp1406I 5 881 2249 3148 4942 5315PstI 1 4961PvuI 1 5086PvuII 3 1819 1912 3414RcaI 4 617 4543 5551 5656RsaI 4 165 1366 3629 5196SapI 1 3707Sau96I 22Sau3AI 37ScaI 1 5196ScrFI 24SfaNI 24SfcI 5 138 465 4088 4279 4957SgrAI 1 538SphI 1 694SspI 1 5520StyI 3 244 392 2717TaqI 14TaqII 8 1127 1345 2018 3725 50645249 5402 5419TfiI 7 1898 2200 2354 2652 28733377 3798ThaI 39TseI 28Tsp45I 9 124 1400 2228 2495 32623475 3570 4972 5183Tsp509I 16Tth111I 1 3568Tth111II 7 1058 1751 3284 4413 44204452 5708UbaJI 24VspI 4 480 1904 1963 4888XbaI 1 431XcmI 3 1075 1591 1609XhoI 1 324XmnI 2 3381 5315Enzymes that do not cut pET-16b:AflII AgeI AscI AvrII BaeIBseRI BsrGI Bsu36I DraIII FseIKpnI MunI NheI NotI NsiINspV PacI PmeI PmlI RleAIRsrII SacI SacII SalI SexAISfiI SgfI SmaI SnaBI SpeISrfI Sse8387I StuI SunI SwaITB046 2/00Novagen • FAX 608-238-1388 • E-MAIL novatech@novagen.com