pBridge
Yeast three-hybrid vector for expressing a "bait" protein fused to the GAL4 DNA-binding domain, plus an additional bridge protein.
Sequence Author: Clontech (TaKaRa)
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Sticky ends from different PfoI sites may not be compatible. |
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Sticky ends from different TsoI sites may not be compatible.After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.For full activity, add fresh S-adenosylmethionine (SAM). |
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Sticky ends from different BglI sites may not be compatible. |
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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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The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible. |
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Sticky ends from different AlwNI sites may not be compatible. |
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Sticky ends from different AflIII sites may not be compatible. |
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PciI is inhibited by nonionic detergents. |
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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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Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present. |
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SmaI can be used at 37°C for brief incubations. |
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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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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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Sticky ends from different BanII sites may not be compatible. |
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Efficient cleavage requires at least two copies of the PaqCI recognition sequence. Sticky ends from different PaqCI sites may not be compatible.Cleavage can be improved with PaqCI Activator. |
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Sticky ends from different BstXI sites may not be compatible. |
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Sticky ends from different Bsu36I sites may not be compatible. |
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Sticky ends from different BstAPI sites may not be compatible. |
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Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present. This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.Sticky ends from different Bpu10I sites may not be compatible. |
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Efficient cleavage requires at least two copies of the NarI recognition sequence. |
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Efficient cleavage requires at least two copies of the PluTI recognition sequence. |
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Sticky ends from different PflMI sites may not be compatible. |
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Prolonged incubation with NdeI may lead to removal of additional nucleotides. |
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Sticky ends from different PpuMI 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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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. |
2μ ori 5363 .. 6527 = 1165 bp yeast 2μ plasmid origin of replication |
2μ ori 5363 .. 6527 = 1165 bp yeast 2μ plasmid origin of replication |
AmpR 4116 .. 4976 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 2: 4116 .. 4907 = 792 bp 263 amino acids = 28.9 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 4116 .. 4976 = 861 bp 286 amino acids = 31.6 kDa 2 segments Segment 1: signal sequence 4908 .. 4976 = 69 bp 23 amino acids = 2.6 kDa Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
AmpR 4116 .. 4976 = 861 bp 286 amino acids = 31.6 kDa 2 segments Product: β-lactamase confers resistance to ampicillin, carbenicillin, and related antibiotics |
TRP1 1161 .. 1835 = 675 bp 224 amino acids = 24.1 kDa Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis yeast auxotrophic marker |
TRP1 1161 .. 1835 = 675 bp 224 amino acids = 24.1 kDa Product: phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis yeast auxotrophic marker |
ori 3357 .. 3945 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
ori 3357 .. 3945 = 589 bp high-copy-number ColE1/pMB1/pBR322/pUC origin of replication |
GAL4 DNA binding domain 434 .. 874 = 441 bp 147 amino acids = 16.9 kDa Product: DNA binding domain of the GAL4 transcriptional activator |
GAL4 DNA binding domain 434 .. 874 = 441 bp 147 amino acids = 16.9 kDa Product: DNA binding domain of the GAL4 transcriptional activator |
ADH1 promoter 10 .. 406 = 397 bp promoter for alcohol dehydrogenase 1 |
ADH1 promoter 10 .. 406 = 397 bp promoter for alcohol dehydrogenase 1 |
MET17 promoter 2259 .. 2609 = 351 bp expression is repressed by methionine |
MET17 promoter 2259 .. 2609 = 351 bp expression is repressed by methionine |
PGK1 terminator 2825 .. 3095 = 271 bp transcription terminator for 3-phosphoglycerate kinase gene |
PGK1 terminator 2825 .. 3095 = 271 bp transcription terminator for 3-phosphoglycerate kinase gene |
ADH1 terminator 921 .. 1108 = 188 bp transcription terminator for alcohol dehydrogenase 1 |
ADH1 terminator 921 .. 1108 = 188 bp transcription terminator for alcohol dehydrogenase 1 |
AmpR promoter 4977 .. 5081 = 105 bp |
AmpR promoter 4977 .. 5081 = 105 bp |
TRP1 promoter 1836 .. 1937 = 102 bp |
TRP1 promoter 1836 .. 1937 = 102 bp |
ATG 2619 .. 2621 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
ATG 2619 .. 2621 = 3 bp 1 amino acid = 149.2 Da Product: start codon |
HA 2631 .. 2657 = 27 bp 9 amino acids = 1.1 kDa Product: HA (human influenza hemagglutinin) epitope tag |
HA 2631 .. 2657 = 27 bp 9 amino acids = 1.1 kDa Product: HA (human influenza hemagglutinin) epitope tag |
SV40 NLS 2679 .. 2699 = 21 bp 7 amino acids = 883.1 Da Product: nuclear localization signal of SV40 large T antigen |
SV40 NLS 2679 .. 2699 = 21 bp 7 amino acids = 883.1 Da Product: nuclear localization signal of SV40 large T antigen |
MCS 1 878 .. 905 = 28 bp multiple cloning site |
MCS 1 878 .. 905 = 28 bp multiple cloning site |
MCS 2 2700 .. 2723 = 24 bp multiple cloning site |
MCS 2 2700 .. 2723 = 24 bp multiple cloning site |
ORF: 1075 .. 1425 = 351 bp ORF: 116 amino acids = 12.7 kDa |
ORF: 4246 .. 4512 = 267 bp ORF: 88 amino acids = 9.2 kDa |
ORF: 434 .. 955 = 522 bp ORF: 173 amino acids = 20.0 kDa |
ORF: 5460 .. 5726 = 267 bp ORF: 88 amino acids = 10.3 kDa |
ORF: 1161 .. 1835 = 675 bp ORF: 224 amino acids = 24.1 kDa |
ORF: 2259 .. 2618 = 360 bp ORF: 119 amino acids = 13.8 kDa |
ORF: 4116 .. 4976 = 861 bp ORF: 286 amino acids = 31.6 kDa |
ORF: 1304 .. 1585 = 282 bp ORF: 93 amino acids = 10.8 kDa |
ORF: 6202 .. 6492 = 291 bp ORF: 96 amino acids = 11.5 kDa |
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