出品公司: | Agilent |
---|---|
载体名称: | pFB |
质粒类型: | 逆病毒载体 |
高拷贝/低拷贝: | 低拷贝 |
克隆方法: | 限制性内切酶,多克隆位点 |
启动子: | -- |
载体大小: | 5117 bp |
5' 测序引物及序列: | CMV Forward: CGCAAATGGGCGGTAGGCGTG |
3' 测序引物及序列: | -- |
载体标签: | -- |
载体抗性: | 氨苄青霉素 |
筛选标记: | 无 |
克隆菌株: | DH5α 等通用菌株 |
宿主细胞(系): | 常规细胞系(293、CV-1、CHO等) |
备注: | -- |
产品目录号: | #217563 |
稳定性: | 稳表达 |
组成型/诱导型: | 组成型 |
病毒/非病毒: | 逆转录病毒 |
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载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
pFB载体描述The Agilent pFB and pFB-Neo plasmid vectors for retroviral gene delivery and expression are derived from the Moloney murine leukemia virus (MMLV) and can be used to produce high titer viral stocks with MMLVbased packaging cell lines.1,2 Both vectors contain the bacterial origin of replication and ampicillin-resistance gene from pBR322, an extended MMLV packaging signal (ψ+), and a multiple cloning site (MCS) located between the MMLV 5′ and 3′ long terminal repeat sequences (LTRs).The pFB vector is a minimal MMLV-based vector that can accommodate larger inserts than pFB-Neo. The pFB vector does not contain any exogenous sequence other than the sequence of the MCS.The pFB-Neo vector differs from the pFB vector only by the presence of a cassette consisting of the internal ribosome entry site (IRES) from the encephalomyocarditis virus (EMCV) and the neomycin-resistance gene (neor). The open reading frame (ORF) for the neor gene is positioned downstream of the MCS and follows the IRES. A dicistronic message encoding the gene of interest and the neor gene is expressed from the viral promoter within the 5′ LTR.pFB载体限制性酶切位点pFB, 5117 bp version 013001Enzymes with 1-10 cleavage sites: #sites -- Bp position of recognition site -- AatII 2 978, 5043 Acc65I 3 647, 1846, 2643 AccI 3 1967, 2057, 3003 AflII 3 202, 1265, 2198 AflIII 2 164, 3232 AgeI 1 1840 AhdI 6 687, 733, 1274, 2683, 2729 4120 Alw44I 4 1054, 3048, 3546, 4792 AlwNI 5 323, 398, 2319, 2394, 3643 ApaI 1 1238 ApoI 3 87, 1128, 2063 AvaI 9 577, 610, 643, 1241, 2018 2074, 2573, 2606, 2639 BamHI 1 2069 BanII 7 580, 593, 764, 1238, 2576 2589, 2760 BbeI 3 615, 1656, 2611 BbsI 1 5110 BbvCI 2 453, 2449 BciVI 5 656, 1986, 2652, 3441, 4968 BglI 1 4239 BglII 1 1678 BpmI 3 1799, 2146, 4210 Bpu10I 7 337, 412, 453, 1548, 2333 2408, 2449 BsaAI 3 1816, 2000, 2984 BsaHI 6 615, 978, 1656, 2611, 4661 5043 BsaI 9 694, 715, 782, 1414, 1802 2690, 2711, 2778, 4192 BsaWI 5 138, 1840, 3438, 3585, 4416 BseRI 5 718, 1068, 1561, 1600, 2714 BsiEI 8 961, 1034, 1838, 2080, 3145 3569, 4492, 4641 BsiHKAI 7 580, 1054, 2576, 3048, 3546 4707, 4792 BsmBI 6 976, 1093, 1337, 1396, 1582 2881 Bsp120I 1 1238 BspHI 3 3952, 4960, 5065 BspLU11I 2 164, 3232 BsrBI 5 1299, 2077, 2102, 3163, 4964 BsrDI 2 4179, 4361 BsrGI 1 1541 BssHII 2 563, 2559 BssSI 3 3405, 4789, 5096 Bst1107I 1 3003 BstAPI 5 322, 397, 2318, 2393, 3050 BstEII 1 1346 BstF5I 9 682, 1386, 1573, 2031, 2678 2907, 4105, 4286, 4573 Bsu36I 1 1276 Cfr10I 2 1840, 4205 Csp6I 8 648, 895, 1542, 1819, 1847 2644, 3039, 4604 DraI 3 3989, 4008, 4700 DraIII 1 1873 DrdI 2 2921, 3334 DsaI 2 151, 931 EaeI 7 827, 961, 1368, 1389, 1668 2080, 4513 EarI 4 1334, 1564, 3110, 4914 Ecl136II 2 580, 2576 Eco52I 2 961, 2080 Eco57I 2 3759, 4807 Eco72I 1 2000 EcoNI 1 1647 EcoO109I 5 499, 1477, 1924, 2495, 5100 EcoRI 1 2063 EcoRV 4 308, 383, 2304, 2379 EheI 3 615, 1656, 2611 FokI 9 682, 1386, 1573, 2031, 2678 2907, 4105, 4286, 4573 FspI 1 4345 HaeII 5 615, 1656, 2611, 3106, 3476 HgaI 6 889, 1166, 2938, 3334, 3912 4662 HincII 2 2057, 4664 HinfI 10 637, 1020, 1045, 1863, 2108 2633, 3132, 3207, 3603, 4120 KasI 3 615, 1656, 2611 KpnI 3 647, 1846, 2643 MaeII 10 979, 1340, 1381, 1817, 2001 2985, 3935, 4351, 4724, 5044 MboII 10 1334, 1565, 2096, 3111, 3882 3973, 4728, 4806, 4915, 5111 MscI 3 827, 1368, 1668 MslI 4 159, 4373, 4532, 4891 MunI 2 11, 21 NarI 3 615, 1656, 2611 NdeI 3 1664, 1672, 3054 NheI 5 6, 16, 26, 197, 2193 NotI 1 2079 NspI 3 164, 2867, 3232 PleI 9 637, 1020, 1045, 1863, 2108 2633, 3132, 3603, 4120 PpuMI 4 499, 1477, 1924, 2495 PshAI 1 1015 Psp1406I 2 4350, 4723 PstI 3 1178, 1360, 4366 PvuI 2 1034, 4492 PvuII 4 286, 361, 2282, 2357 RsaI 8 648, 895, 1542, 1819, 1847 2644, 3039, 4604 SacI 2 580, 2576 SacII 1 151 SalI 1 2057 SapI 1 3109 ScaI 1 4603 SchI 9 637, 1020, 1045, 1863, 2108 2633, 3132, 3603, 4120 SexAI 1 1474 SfcI 7 182, 1178, 1360, 2178, 3497 3688, 4366 SmaI 4 643, 1241, 2018, 2639 SmlI 8 202, 1265, 2074, 2198, 3338 3600, 3877, 4745 SpeI 1 897 SrfI 1 1240 SspI 1 4927 StyI 5 496, 706, 1504, 2492, 2702 TaiI 10 979, 1340, 1381, 1817, 2001 2985, 3935, 4351, 4724, 5044 TaqI 9 1297, 1608, 1619, 2050, 2058 2075, 2119, 3332, 4776 TatI 3 1541, 3038, 4603 TfiI 1 3207 Tsp45I 6 1282, 1491, 2887, 2982, 4382 4593 TspRI 10 1284, 1875, 3128, 3634, 3647 3918, 4067, 4172, 4519, 4546 Tth111I 4 633, 1473, 2629, 2976 VspI 1 4296 XbaI 3 464, 2036, 2460 XhoI 1 2074 XhoII 8 1678, 2069, 3873, 3884, 3970 3982, 4750, 4767 XmaI 4 643, 1241, 2018, 2639 XmnI 1 4720Enzymes that do NOT cut molecule:AccIII AscI BclI BlnI Bpu1102I BsaBI BsgI BsiWI BsmI BspMI BstBI BstXI ClaI Eco47III FseI HindIII HpaI MluI NaeI NcoI NgoMIV NruI NsiI PacI PmeI Ppu10I RsrII SanDI SbfI SfiI SgfI SgrAI SnaBI SphI StuI SwaI Van91I XcmI
载体序列
pFB, 5117 bp version 013001 NOTE: The following sequence has been verified for accuracy at the junctions. The remainder of the sequence has been obtained from existing data. 1 GAATTGCTAG CAATTGCTAG CAATTGCTAG CAATTCATAC CAGATCACCG 51 AAAACTGTCC TCCAAATGTG TCCCCCTCAC ACTCCCAAAT TCGCGGGCTT 101 CTGCCTCTTA GACCACTCTA CCCTATTCCC CACACTCACC GGAGCCAAAG 151 CCGCGGGACA TATACATGTG AAAGACCCCA CCTGTAGGTT TGGCAAGCTA 201 GCTTAAGTAA CGCCATTTTG CAAGGCATGG AAAAATACAT AACTGAGAAT 251 AGAAAAGTTC AGATCAAGGT CAGGAACAGA TGGAACAGCT GAATATGGGC 301 CAAAGCGGAT ATCTGTGGTA AGCAGTTCCT GCCCCGGCTC AGGGCCAAGA 351 ACAGATGGAA CAGCTGAATA TGGGCCAAAC AGGATATCTG TGGTAAGCAG 401 TTCCTGCCCC GGCTCAGGGC CAAGAACAGA TGGTCCCCAG ATGCGGTCCA 451 GCCCTCAGCA GTTTCTAGAG AACCATCAGA TGTTTCCAGG GTGCCCCAAG 501 GACCTGAAAT GACCCTGTGC CTTATTTGAA CTAACCAATC AGTTCGCTTC 551 TCGCTTCTGT TCGCGCGCTT CTGCTCCCCG AGCTCAATAA AAGAGCCCAC 601 AACCCCTCAC TCGGGGCGCC AGTCCTCCGA TTGACTGAGT CGCCCGGGTA 651 CCCGTGTATC CAATAAACCC TCTTGCAGTT GCATCCGACT TGTGGTCTCG 701 CTGTTCCTTG GGAGGGTCTC CTCTGAGTGA TTGACTACCC GTCAGCGGGG 751 GTCTTTCATT TGGGGGCTCG TCCGGGATCG GGAGACCCCT GCCCAGGGAC 801 CACCGACCCA CCACCGGGAG GTAAGCTGGC CAGCAACTTA TCTGTGTCTG 851 TCCGATTGTC TAGTGTCTAT GACTGATTTT ATGCGCCTGC GTCGGTACTA 901 GTTAGCTAAC TAGCTCTGTA TCTGGCGGAC CCGTGGTGGA ACTGACGAGT 951 TCGGAACACC CGGCCGCAAC CCTGGGAGAC GTCCCAGGGA CTTCGGGGGC 1001 CGTTTTTGTG GCCCGACCTG AGTCCAAAAA TCCCGATCGT TTTGGACTCT 1051 TTGGTGCACC CCCCTTAGAG GAGGGATATG TGGTTCTGGT AGGAGACGAG 1101 AACCTAAAAC AGTTCCCGCC TCCGTCTGAA TTTTTGCTTT CGGTTTGGGA 1151 CCGAAGCCGC GCCGCGCGTC TTGTCTGCTG CAGCATCGTT CTGTGTTGTC 1201 TCTGTCTGAC TGTGTTTCTG TATTTGTCTG AAAATATGGG CCCGGGCCAG 1251 ACTGTTACCA CTCCCTTAAG TTTGACCTTA GGTCACTGGA AAGATGTCGA 1301 GCGGATCGCT CACAACCAGT CGGTAGATGT CAAGAAGAGA CGTTGGGTTA 1351 CCTTCTGCTC TGCAGAATGG CCAACCTTTA ACGTCGGATG GCCGCGAGAC 1401 GGCACCTTTA ACCGAGACCT CATCACCCAG GTTAAGATCA AGGTCTTTTC 1451 ACCTGGCCCG CATGGACACC CAGACCAGGT CCCCTACATC GTGACCTGGG 1501 AAGCCTTGGC TTTTGACCCC CCTCCCTGGG TCAAGCCCTT TGTACACCCT 1551 AAGCCTCCGC CTCCTCTTCC TCCATCCGCC CCGTCTCTCC CCCTTGAACC 1601 TCCTCGTTCG ACCCCGCCTC GATCCTCCCT TTATCCAGCC CTCACTCCTT 1651 CTCTAGGCGC CCCCATATGG CCATATGAGA TCTTATATGG GGCACCCCCG 1701 CCCCTTGTAA ACTTCCCTGA CCCTGACATG ACAAGAGTTA CTAACAGCCC 1751 CTCTCTCCAA GCTCACTTAC AGGCTCTCTA CTTAGTCCAG CACGAAGTCT 1801 GGAGACCTCT GGCGGCACGT ACCAAGAACA ACTGGACCGA CCGGTGGTAC 1851 CTCACCCTTA CCGAGTCGGC GACACAGTGT GGGTCCGCCG ACACCAGACT 1901 AAGAACCTAG AACCTCGCTG GAAAGGACCT TACACAGTCC TGCTGACCAC 1951 CCCCACCGCC CTCAAAGTAG ACGGCATCGC AGCTTGGATA CACGCCGCCC 2001 ACGTGAAGGC TGCCGACCCC GGGGGTGGAC CATCCTCTAG ACTGCCGGAT 2051 CGAATTGTCG ACGAATTCGG ATCCTCGAGC GGCCGCGATC CGAGTTCTTC 2101 TGAGCGGGAC TCTGGGGTTC GATAAAATAA AAGATTTTAT TTAGTCTCCA 2151 GAAAAAGGGG GGAATGAAAG ACCCCACCTG TAGGTTTGGC AAGCTAGCTT 2201 AAGTAACGCC ATTTTGCAAG GCATGGAAAA ATACATAACT GAGAATAGAG 2251 AAGTTCAGAT CAAGGTCAGG AACAGATGGA ACAGCTGAAT ATGGGCCAAA 2301 CAGGATATCT GTGGTAAGCA GTTCCTGCCC CGGCTCAGGG CCAAGAACAG 2351 ATGGAACAGC TGAATATGGG CCAAACAGGA TATCTGTGGT AAGCAGTTCC 2401 TGCCCCGGCT CAGGGCCAAG AACAGATGGT CCCCAGATGC GGTCCAGCCC 2451 TCAGCAGTTT CTAGAGAACC ATCAGATGTT TCCAGGGTGC CCCAAGGACC 2501 TGAAATGACC CTGTGCCTTA TTTGAACTAA CCAATCAGTT CGCTTCTCGC 2551 TTCTGTTCGC GCGCTTCTGC TCCCCGAGCT CAATAAAAGA GCCCACAACC 2601 CCTCACTCGG GGCGCCAGTC CTCCGATTGA CTGAGTCGCC CGGGTACCCG 2651 TGTATCCAAT AAACCCTCTT GCAGTTGCAT CCGACTTGTG GTCTCGCTGT 2701 TCCTTGGGAG GGTCTCCTCT GAGTGATTGA CTACCCGTCA GCGGGGGTCT 2751 TTCATTTGGG GGCTCGTCCG GGATCGGGAG ACCCCTGCCC AGGGACCACC 2801 GACCCACCAC CGGGAGGTAA GCTGGCTGCC TCGCGCGTTT CGGTGATGAC 2851 GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA CAGCTTGTCT 2901 GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG TCAGCGGGTG 2951 TTGGCGGGTG TCGGGGCGCA GCCATGACCC AGTCACGTAG CGATAGCGGA 3001 GTGTATACTG GCTTAACTAT GCGGCATCAG AGCAGATTGT ACTGAGAGTG 3051 CACCATATGC GGTGTGAAAT ACCGCACAGA TGCGTAAGGA GAAAATACCG 3101 CATCAGGCGC TCTTCCGCTT CCTCGCTCAC TGACTCGCTG CGCTCGGTCG 3151 TTCGGCTGCG GCGAGCGGTA TCAGCTCACT CAAAGGCGGT AATACGGTTA 3201 TCCACAGAAT CAGGGGATAA CGCAGGAAAG AACATGTGAG CAAAAGGCCA 3251 GCAAAAGGCC AGGAACCGTA AAAAGGCCGC GTTGCTGGCG TTTTTCCATA 3301 GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG 3351 TGGCGAAACC CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG 3401 CTCCCTCGTG CGCTCTCCTG TTCCGACCCT GCCGCTTACC GGATACCTGT 3451 CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC TTTCTCATAG CTCACGCTGT 3501 AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA 3551 CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC 3601 TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT 3651 GGTAACAGGA TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT 3701 GAAGTGGTGG CCTAACTACG GCTACACTAG AAGGACAGTA TTTGGTATCT 3751 GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG TAGCTCTTGA 3801 TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA 3851 GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGATCCT TTGATCTTTT 3901 CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTG 3951 GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA 4001 ATGAAGTTTT AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA 4051 GTTACCAATG CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT 4101 CGTTCATCCA TAGTTGCCTG ACTCCCCGTC GTGTAGATAA CTACGATACG 4151 GGAGGGCTTA CCATCTGGCC CCAGTGCTGC AATGATACCG CGAGACCCAC 4201 GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC CGGAAGGGCC 4251 GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA 4301 TTGTTGCCGG GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA 4351 ACGTTGTTGC CATTGCTGCA GGCATCGTGG TGTCACGCTC GTCGTTTGGT 4401 ATGGCTTCAT TCAGCTCCGG TTCCCAACGA TCAAGGCGAG TTACATGATC 4451 CCCCATGTTG TGCAAAAAAG CGGTTAGCTC CTTCGGTCCT CCGATCGTTG 4501 TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT GGCAGCACTG 4551 CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG 4601 TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT 4651 GCTCTTGCCC GGCGTCAACA CGGGATAATA CCGCGCCACA TAGCAGAACT 4701 TTAAAAGTGC TCATCATTGG AAAACGTTCT TCGGGGCGAA AACTCTCAAG 4751 GATCTTACCG CTGTTGAGAT CCAGTTCGAT GTAACCCACT CGTGCACCCA 4801 ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG GTGAGCAAAA 4851 ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG 4901 TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG 4951 GTTATTGTCT CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA 5001 CAAATAGGGG TTCCGCGCAC ATTTCCCCGA AAAGTGCCAC CTGACGTCTA 5051 AGAAACCATT ATTATCATGA CATTAACCTA TAAAAATAGG CGTATCACGA 5101 GGCCCTTTCG TCTTCAA
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