pFB-Neo載體描述 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載體限制性酶切位點 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
