pFB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).
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載體限制性酶切位點 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
