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pFB-Neo载体质粒基本信息
pFB-Neo质粒图谱载体图谱和pFB-Neo载体序列质粒序列多克隆位点信息
pFB-Neo质粒载体简介
pFB-Neo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), see Figures 1 and 2.
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-Neo载体限制性酶切位点
pFB-Neo, 6572 bp version 037004
Enzymes with 1-10 cleavage sites:
#sites -- Bp position of recognition site --
AarI 1 2348
AatII 2 978, 6498
Acc65I 4 647, 1847, 2460, 4098
AccI 3 1968, 2058, 4458
AclI 2 5805, 6178
AcuI 6 2443, 2613, 3017, 3449, 5214, 6262
AflII 3 202, 1265, 3653
AflIII 4 164, 2334, 2509, 4687
AgeI 1 1841
AhdI 6 687, 733, 1274, 4138, 4184, 5575
AleI 1 2658
Alw44I 5 1054, 2496, 4503, 5001, 6247
AlwNI 5 323, 398, 3774, 3849, 5098
ApaI 2 1238, 2132
ApoI 3 87, 1128, 2064
AseI 1 5751
AvaI 9 577, 610, 643, 1241, 2019, 2075
4028, 4061, 4094
BamHI 1 2070
BanII 9 580, 593, 764, 1238, 2132, 3253
4031, 4044, 4215
BbeI 4 615, 1656, 2891, 4066
BbsI 4 2152, 2252, 2618, 6565
BbvCI 2 453, 3904
BceAI 7 999, 1399, 1972, 2112, 2866, 3302
5173
BciVI 6 656, 1987, 3100, 4107, 4896, 6423
BglI 2 2315, 5694
BglII 1 1678
BlnI 1 2170
BmgBI 1 2559
BmrI 4 2369, 2831, 4433, 5625
BmtI 5 6, 16, 26, 197, 3648
BpmI 3 1799, 3601, 5665
Bpu10I 7 337, 412, 453, 1548, 3788, 3863
3904
BpuEI 6 2421, 3283, 4793, 5055, 5332, 6200
BsaAI 4 2001, 2333, 3194, 4439
BsaHI 7 615, 978, 1656, 2891, 4066, 6116
6498
BsaI 9 694, 715, 782, 1414, 1802, 4145
4166, 4233, 5647
BsaWI 7 138, 1841, 2090, 2923, 4893, 5040
5871
BseRI 7 718, 1068, 1561, 1600, 2418, 2670
4169
BseYI 2 3396, 4991
BsgI 1 2719
BsiEI 10 961, 1034, 1839, 2081, 2583, 2798
4600, 5024, 5947, 6096
BsiHKAI 10 580, 1054, 2496, 3001, 3191, 4031
4503, 5001, 6162, 6247
BsmBI 6 976, 1093, 1337, 1396, 1582, 4336
BsmI 2 2165, 2198
BspHI 3 5407, 6415, 6520
BspMI 3 2349, 2788, 3151
BsrBI 6 2078, 2644, 3503, 3557, 4618, 6419
BsrDI 4 2123, 3118, 5634, 5816
BsrGI 1 1541
BssHII 3 563, 3289, 4014
BssSI 4 3484, 4860, 6244, 6551
Bst1107I 1 4458
BstAPI 5 322, 397, 3773, 3848, 4505
BstEII 1 1346
Bsu36I 1 1276
BtgI 6 151, 931, 2552, 2702, 2747, 3324
BtgZI 4 1976, 3178, 3329, 3522
BtsI 2 5973, 6001
Cfr10I 4 1841, 3211, 3392, 5660
ClaI 1 2585
Csp6I 9 648, 895, 1542, 1848, 2461, 3197
4099, 4494, 6059
DraI 3 5444, 5463, 6155
DraIII 2 1874, 2377
DrdI 3 2913, 4376, 4789
EagI 3 961, 2081, 2798
EarI 6 1334, 1564, 3241, 3451, 4565, 6369
EciI 8 924, 1556, 1575, 1885, 3405, 4759
4905, 5733
Eco57MI 9 1799, 2443, 2613, 3017, 3449, 3601
5214, 5665, 6262
EcoICRI 2 580, 4031
EcoNI 1 1647
EcoO109I 8 499, 1477, 1925, 2131, 2487, 2540
3950, 6555
EcoRI 1 2064
EcoRV 4 308, 383, 3759, 3834
FspI 2 2992, 5800
HaeII 6 615, 1656, 2891, 4066, 4561, 4931
HgaI 6 889, 1166, 4393, 4789, 5367, 6117
HincII 2 2058, 6119
HindIII 1 2243
KasI 4 615, 1656, 2891, 4066
KpnI 4 647, 1847, 2460, 4098
MlyI 10 637, 1020, 1045, 1864, 2405, 3563
4088, 4587, 5058, 5575
MmeI 6 684, 1383, 2387, 4135, 4877, 5061
MscI 4 827, 1368, 1668, 2972
MslI 6 159, 2658, 3325, 5828, 5987, 6346
MunI 2 11, 21
NaeI 1 3392
NarI 4 615, 1656, 2891, 4066
NcoI 3 2702, 2747, 3324
NdeI 3 1664, 1672, 4509
NgoMIV 1 3392
NheI 5 6, 16, 26, 197, 3648
NotI 1 2080
NspI 6 164, 2500, 2509, 3293, 4322, 4687
PciI 3 164, 2509, 4687
PfoI 3 771, 4222, 4331
PleI 10 637, 1020, 1045, 1864, 2405, 3563
4088, 4587, 5058, 5575
PmlI 2 2001, 2333
PpuMI 4 499, 1477, 1925, 3950
PshAI 1 1015
PspOMI 2 1238, 2132
PstI 4 1178, 1360, 2941, 5821
PvuI 3 1034, 2583, 5947
PvuII 5 286, 361, 2996, 3737, 3812
RsaI 9 648, 895, 1542, 1848, 2461, 3197
4099, 4494, 6059
RsrII 1 3407
SacI 2 580, 4031
SacII 1 151
SalI 1 2058
SapI 3 3241, 3451, 4564
ScaI 1 6058
SexAI 1 1474
SfcI 9 182, 1178, 1360, 2267, 2941, 3633
4952, 5143, 5821
SfoI 4 615, 1656, 2891, 4066
SmaI 4 643, 1241, 2019, 4094
SmlI 10 202, 1265, 2075, 2421, 3283, 3653
4793, 5055, 5332, 6200
SpeI 1 897
SphI 1 3293
SrfI 1 1240
SspI 1 6382
StyI 9 496, 706, 1504, 2170, 2702, 2747
3324, 3947, 4157
TatI 3 1541, 4493, 6058
TfiI 3 3377, 3511, 4662
Tsp45I 8 1282, 1491, 3013, 3319, 4342, 4437
5837, 6048
TspDTI 9 34, 755, 2936, 3379, 3619, 4206
5456, 5558, 5861
TspGWI 3 1121, 6030, 6347
Tth111I 5 633, 1473, 3007, 4084, 4431
Van91I 1 2466
XbaI 3 464, 2037, 3915
XhoI 1 2075
XmaI 4 643, 1241, 2019, 4094
XmnI 2 2227, 6175
ZraI 2 978, 6498
Enzymes that do NOT cut molecule:
AccIII AscI AsiSI BclI BfrBI
BlpI BsaBI BsiWI BstBI BstXI
Eco47III FseI FspAI HpaI MluI
NruI NsiI PacI PmeI PsiI
SanDI SbfI SfiI SgrAI SnaBI
StuI SwaI XcmI
pFB-Neo质粒序列载体序列
pFB-Neo, 6572 bp version 037003
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 GCAGATCGCT 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 GGCGGCAGCC TACCAAGAAC AACTGGACCG ACCGGTGGTA
1851 CCTCACCCTT ACCGAGTCGG CGACACAGTG TGGGTCCGCC GACACCAGAC
1901 TAAGAACCTA GAACCTCGCT GGAAAGGACC TTACACAGTC CTGCTGACCA
1951 CCCCCACCGC CCTCAAAGTA GACGGCATCG CAGCTTGGAT ACACGCCGCC
2001 CACGTGAAGG CTGCCGACCC CGGGGGTGGA CCATCCTCTA GACTGCCGGA
2051 TCGAATTGTC GACGAATTCG GATCCTCGAG CGGCCGCGAT CCGGTTATTT
2101 TCCACCATAT TGCCGTCTTT TGGCAATGTG AGGGCCCGGA AACCTGGCCC
2151 TGTCTTCTTG ACGAGCATTC CTAGGGGTCT TTCCCCTCTC GCCAAAGGAA
2201 TGCAAGGTCT GTTGAATGTC GTGAAGGAAG CAGTTCCTCT GGAAGCTTCT
2251 TGAAGACAAA CAACGTCTGT AGCGACCCTT TGCAGGCAGC GGAACCCCCC
2301 ACCTGGCGAC AGGTGCCTCT GCGGCCAAAA GCCACGTGTA TAAGATACAC
2351 CTGCAAAGGC GGCACAACCC CAGTGCCACG TTGTGAGTTG GATAGTTGTG
2401 GAAAGAGTCA AATGGCTCTC CTCAAGCGTA TTCAACAAGG GGCTGAAGGA
2451 TGCCCAGAAG GTACCCCATT GTATGGGATC TGATCTGGGG CCTCGGTGCA
2501 CATGCTTTAC ATGTGTTTAG TCGAGGTTAA AAAACGTCTA GGCCCCCCGA
2551 ACCACGGGGA CGTGGTTTTC CTTTGAAAAA CACGATCGAT AATATGGAAC
2601 AAAAACTTAT TTCTGAAGAA GACTTGGACA CCAAACTTTC CTGCCGCTCG
2651 ATTTCTCCAC CCAGGTGAAC TCCTCCCTCA CCTCCCCGAC GGGGCGAGGC
2701 TCCATGGCTG CCCCCTCGCT GCACCCGTCC CTGGTGCCAC GCGGTTCCAT
2751 GGGATCGTTT CGCATGATTG AACAAGATGG ATTGCACGCA GGTTCTCCGG
2801 CCGCTTGGGT GGAGAGGCTA TTCGGCTATG ACTGGGCACA ACAGACAATC
2851 GGCTGCTCTG ATGCCGCCGT GTTCCGGCTG TCAGCGCAGG GGCGCCCGGT
2901 TCTTTTTGTC AAGACCGACC TGTCCGGTGC CCTGAATGAA CTGCAGGACG
2951 AGGCAGCGCG GCTATCGTGG CTGGCCACGA CGGGCGTTCC TTGCGCAGCT
3001 GTGCTCGACG TTGTCACTGA AGCGGGAAGG GACTGGCTGC TATTGGGCGA
3051 AGTGCCGGGG CAGGATCTCC TGTCATCTCA CCTTGCTCCT GCCGAGAAAG
3101 TATCCATCAT GGCTGATGCA ATGCGGCGGC TGCATACGCT TGATCCGGCT
3151 ACCTGCCCAT TCGACCACCA AGCGAAACAT CGCATCGAGC GAGCACGTAC
3201 TCGGATGGAA GCCGGTCTTG TCGATCAGGA TGATCTGGAC GAAGAGCATC
3251 AGGGGCTCGC GCCAGCCGAA CTGTTCGCCA GGCTCAAGGC GCGCATGCCC
3301 GACGGCGAGG ATCTCGTCGT GACCCATGGC GATGCCTGCT TGCCGAATAT
3351 CATGGTGGAA AATGGCCGCT TTTCTGGATT CATCGACTGT GGCCGGCTGG
3401 GTGTGGCGGA CCGCTATCAG GACATAGCGT TGGCTACCCG TGATATTGCT
3451 GAAGAGCTTG GCGGCGAATG GGCTGACCGC TTCCTCGTGC TTTACGGTAT
3501 CGCCGCTCCC GATTCGCAGC GCATCGCCTT CTATCGCCTT CTTGACGAGT
3551 TCTTCTGAGC GGGACTCTGG GGTTCGATAA AATAAAAGAT TTTATTTAGT
3601 CTCCAGAAAA AGGGGGGAAT GAAAGACCCC ACCTGTAGGT TTGGCAAGCT
3651 AGCTTAAGTA ACGCCATTTT GCAAGGCATG GAAAAATACA TAACTGAGAA
3701 TAGAGAAGTT CAGATCAAGG TCAGGAACAG ATGGAACAGC TGAATATGGG
3751 CCAAACAGGA TATCTGTGGT AAGCAGTTCC TGCCCCGGCT CAGGGCCAAG
3801 AACAGATGGA ACAGCTGAAT ATGGGCCAAA CAGGATATCT GTGGTAAGCA
3851 GTTCCTGCCC CGGCTCAGGG CCAAGAACAG ATGGTCCCCA GATGCGGTCC
3901 AGCCCTCAGC AGTTTCTAGA GAACCATCAG ATGTTTCCAG GGTGCCCCAA
3951 GGACCTGAAA TGACCCTGTG CCTTATTTGA ACTAACCAAT CAGTTCGCTT
4001 CTCGCTTCTG TTCGCGCGCT TCTGCTCCCC GAGCTCAATA AAAGAGCCCA
4051 CAACCCCTCA CTCGGGGCGC CAGTCCTCCG ATTGACTGAG TCGCCCGGGT
4101 ACCCGTGTAT CCAATAAACC CTCTTGCAGT TGCATCCGAC TTGTGGTCTC
4151 GCTGTTCCTT GGGAGGGTCT CCTCTGAGTG ATTGACTACC CGTCAGCGGG
4201 GGTCTTTCAT TTGGGGGCTC GTCCGGGATC GGGAGACCCC TGCCCAGGGA
4251 CCACCGACCC ACCACCGGGA GGTAAGCTGG CTGCCTCGCG CGTTTCGGTG
4301 ATGACGGTGA AAACCTCTGA CACATGCAGC TCCCGGAGAC GGTCACAGCT
4351 TGTCTGTAAG CGGATGCCGG GAGCAGACAA GCCCGTCAGG GCGCGTCAGC
4401 GGGTGTTGGC GGGTGTCGGG GCGCAGCCAT GACCCAGTCA CGTAGCGATA
4451 GCGGAGTGTA TACTGGCTTA ACTATGCGGC ATCAGAGCAG ATTGTACTGA
4501 GAGTGCACCA TATGCGGTGT GAAATACCGC ACAGATGCGT AAGGAGAAAA
4551 TACCGCATCA GGCGCTCTTC CGCTTCCTCG CTCACTGACT CGCTGCGCTC
4601 GGTCGTTCGG CTGCGGCGAG CGGTATCAGC TCACTCAAAG GCGGTAATAC
4651 GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT GTGAGCAAAA
4701 GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT
4751 CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC
4801 AGAGGTGGCG AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT
4851 GGAAGCTCCC TCGTGCGCTC TCCTGTTCCG ACCCTGCCGC TTACCGGATA
4901 CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT GGCGCTTTCT CATAGCTCAC
4951 GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA GCTGGGCTGT
5001 GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA
5051 TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG
5101 CCACTGGTAA CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG
5151 TTCTTGAAGT GGTGGCCTAA CTACGGCTAC ACTAGAAGGA CAGTATTTGG
5201 TATCTGCGCT CTGCTGAAGC CAGTTACCTT CGGAAAAAGA GTTGGTAGCT
5251 CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT TTTTGTTTGC
5301 AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT
5351 CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA
5401 TTTTGGTCAT GAGATTATCA AAAAGGATCT TCACCTAGAT CCTTTTAAAT
5451 TAAAAATGAA GTTTTAAATC AATCTAAAGT ATATATGAGT AAACTTGGTC
5501 TGACAGTTAC CAATGCTTAA TCAGTGAGGC ACCTATCTCA GCGATCTGTC
5551 TATTTCGTTC ATCCATAGTT GCCTGACTCC CCGTCGTGTA GATAACTACG
5601 ATACGGGAGG GCTTACCATC TGGCCCCAGT GCTGCAATGA TACCGCGAGA
5651 CCCACGCTCA CCGGCTCCAG ATTTATCAGC AATAAACCAG CCAGCCGGAA
5701 GGGCCGAGCG CAGAAGTGGT CCTGCAACTT TATCCGCCTC CATCCAGTCT
5751 ATTAATTGTT GCCGGGAAGC TAGAGTAAGT AGTTCGCCAG TTAATAGTTT
5801 GCGCAACGTT GTTGCCATTG CTGCAGGCAT CGTGGTGTCA CGCTCGTCGT
5851 TTGGTATGGC TTCATTCAGC TCCGGTTCCC AACGATCAAG GCGAGTTACA
5901 TGATCCCCCA TGTTGTGCAA AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT
5951 CGTTGTCAGA AGTAAGTTGG CCGCAGTGTT ATCACTCATG GTTATGGCAG
6001 CACTGCATAA TTCTCTTACT GTCATGCCAT CCGTAAGATG CTTTTCTGTG
6051 ACTGGTGAGT ACTCAACCAA GTCATTCTGA GAATAGTGTA TGCGGCGACC
6101 GAGTTGCTCT TGCCCGGCGT CAACACGGGA TAATACCGCG CCACATAGCA
6151 GAACTTTAAA AGTGCTCATC ATTGGAAAAC GTTCTTCGGG GCGAAAACTC
6201 TCAAGGATCT TACCGCTGTT GAGATCCAGT TCGATGTAAC CCACTCGTGC
6251 ACCCAACTGA TCTTCAGCAT CTTTTACTTT CACCAGCGTT TCTGGGTGAG
6301 CAAAAACAGG AAGGCAAAAT GCCGCAAAAA AGGGAATAAG GGCGACACGG
6351 AAATGTTGAA TACTCATACT CTTCCTTTTT CAATATTATT GAAGCATTTA
6401 TCAGGGTTAT TGTCTCATGA GCGGATACAT ATTTGAATGT ATTTAGAAAA
6451 ATAAACAAAT AGGGGTTCCG CGCACATTTC CCCGAAAAGT GCCACCTGAC
6501 GTCTAAGAAA CCATTATTAT CATGACATTA ACCTATAAAA ATAGGCGTAT
6551 CACGAGGCCC TTTCGTCTTC AA
