pFastBac Dual
Dual-promoter vector for co-expression of two proteins in insect cells using the Bac-to-Bac® baculovirus system.
Sequence Author: Thermo Fisher (Invitrogen)
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BssHII is typically used at 50°C, but is 75% active at 37°C. |
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Efficient cleavage requires at least two copies of the RsrII recognition sequence. Sticky ends from different RsrII sites may not be compatible.For full activity, add fresh DTT. |
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After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility. |
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Sticky ends from different BbsI sites may not be compatible.BbsI gradually loses activity when stored at -20°C. |
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SmaI can be used at 37°C for brief incubations. |
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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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PaeR7I does not recognize the sequence CTCTCGAG. |
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BsrGI is typically used at 37°C, but is even more active at 60°C. |
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The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible. |
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The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible. |
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Sticky ends from different BsmBI sites may not be compatible.BsmBI-v2 is an improved version of BsmBI. |
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Sticky ends from different Esp3I sites may not be compatible. |
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Sticky ends from different BseRI sites may not be compatible.BseRI quickly loses activity at 37°C.Prolonged incubation with BseRI may lead to degradation of the DNA. |
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EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage. |
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Efficient cleavage requires at least two copies of the SacII recognition sequence. |
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Sticky ends from different BstXI sites may not be compatible. |
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Efficient cleavage requires at least two copies of the NgoMIV recognition sequence. |
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Efficient cleavage requires at least two copies of the NaeI recognition sequence. |
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BsaHI is typically used at 37°C, but is even more active at 60°C. |
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Efficient cleavage requires at least two copies of the BpmI recognition sequence. Sticky ends from different BpmI sites may not be compatible.After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.BpmI quickly loses activity at 37°C. |
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Sticky ends from different AlwNI sites may not be compatible. |
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After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility. |
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Sticky ends from different BspQI sites may not be compatible. |
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Sticky ends from different SapI sites may not be compatible.SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot. |
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AmpR 689 .. 1549 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 689 .. 757 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 689 .. 1549 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 758 .. 1549 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 689 .. 1549 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
ori 1720 .. 2308 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ori 1720 .. 2308 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
GmR 2902 .. 3435 = 534 bp 177 amino acids = 19.4 kDa Product: gentamycin acetyltransferase confers resistance to gentamycin |
GmR 2902 .. 3435 = 534 bp 177 amino acids = 19.4 kDa Product: gentamycin acetyltransferase confers resistance to gentamycin |
f1 ori 102 .. 557 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
f1 ori 102 .. 557 = 456 bp f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis |
Tn7R 2611 .. 2835 = 225 bp mini-Tn7 element (right end of the Tn7 transposon) |
Tn7R 2611 .. 2835 = 225 bp mini-Tn7 element (right end of the Tn7 transposon) |
Tn7L 4991 .. 5156 = 166 bp mini-Tn7 element (left end of the Tn7 transposon) |
Tn7L 4991 .. 5156 = 166 bp mini-Tn7 element (left end of the Tn7 transposon) |
SV40 poly(A) signal 4828 .. 4962 = 135 bp SV40 polyadenylation signal |
SV40 poly(A) signal 4828 .. 4962 = 135 bp SV40 polyadenylation signal |
p10 promoter 4345 .. 4454 = 110 bp baculovirus promoter for expression in insect cells |
p10 promoter 4345 .. 4454 = 110 bp baculovirus promoter for expression in insect cells |
AmpR promoter 584 .. 688 = 105 bp |
AmpR promoter 584 .. 688 = 105 bp |
MCS 4606 .. 4704 = 99 bp multiple cloning site |
MCS 4606 .. 4704 = 99 bp multiple cloning site |
polyhedrin promoter 4478 .. 4569 = 92 bp promoter for the baculovirus polyhedrin gene |
polyhedrin promoter 4478 .. 4569 = 92 bp promoter for the baculovirus polyhedrin gene |
MCS 4274 .. 4337 = 64 bp multiple cloning site |
MCS 4274 .. 4337 = 64 bp multiple cloning site |
HSV TK poly(A) signal 4161 .. 4209 = 49 bp herpesvirus thymidine kinase polyadenylation signal |
HSV TK poly(A) signal 4161 .. 4209 = 49 bp herpesvirus thymidine kinase polyadenylation signal |
Pc promoter 3624 .. 3652 = 29 bp 3 segments Segment 3: -10 3624 .. 3629 = 6 bp class 1 integron promoter |
Pc promoter 3624 .. 3652 = 29 bp 3 segments Segment 2: 3630 .. 3646 = 17 bp class 1 integron promoter |
Pc promoter 3624 .. 3652 = 29 bp 3 segments Segment 1: -35 3647 .. 3652 = 6 bp class 1 integron promoter |
Pc promoter 3624 .. 3652 = 29 bp 3 segments class 1 integron promoter |
ORF: 3040 .. 3372 = 333 bp ORF: 110 amino acids = 11.9 kDa |
ORF: 3646 .. 3888 = 243 bp ORF: 80 amino acids = 9.1 kDa |
ORF: 3895 .. 4215 = 321 bp ORF: 106 amino acids = 11.7 kDa |
ORF: 689 .. 1549 = 861 bp ORF: 286 amino acids = 31.6 kDa |
ORF: 2745 .. 3047 = 303 bp ORF: 100 amino acids = 11.8 kDa |
ORF: 3537 .. 3992 = 456 bp ORF: 151 amino acids = 17.4 kDa |
ORF: 1153 .. 1419 = 267 bp ORF: 88 amino acids = 9.2 kDa |
ORF: 2902 .. 3435 = 534 bp ORF: 177 amino acids = 19.4 kDa |
ORF: 3627 .. 3935 = 309 bp ORF: 102 amino acids = 11.5 kDa |
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