| 出品公司: | ZYbscience |
|---|---|
| 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: | ZY217563 |
| 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 |
- ¥3000
- ZYbscience
- 中国/美国
- ZY217563
- 2025年07月10日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pFB
- 库存：
20
- 供应商：
泽叶生物
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 013001
Enzymes 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 4720
Enzymes 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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文献和实验相关实验
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zhangyuxianggx 各位大侠好！现有pcDNA3-mCherry和pGEM-hgene两个东西在滤纸上，老师就说让构建载体，具体是什么意思啊？我理解的是一个真核表达载体和构建好的pGEM质粒，但是不知下一步该怎么做？恳请各位好心的大侠指点一二。 shylook 直接插入cDNA吧 zhangyuxianggx 那个pGEM就是已经插入了DNA的载体。请问这两个东西溶下来以后
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pFB载体
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