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Plasmid Files

pBridge™

Yeast three-hybrid vector for expressing a "bait" protein fused to the GAL4 DNA-binding domain, plus an additional bridge protein.

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pBridge Sequence and MappBridge.dna
Map and Sequence File   
Sequence Author:  Clontech
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 NsiI (6241) PfoI (5222) AatII (5111) ZraI (5109) ScaI (4669) TsoI (4588) PvuI (4559) BglI (4309) BpmI (4259) AhdI (4189) AlwNI (3712) AflIII - PciI (3296) PaeR7I - XhoI (651) HpaI (711) BsrGI (724) EcoRI (878) TspMI - XmaI (883) SmaI (885) BamHI (888) SalI (894) PstI (904) PsiI (963) NgoMIV (1112) NaeI (1114) BanII (1331) AarI (1398) BstXI (1541) Bsu36I (1590) XbaI (1747) BstAPI (1759) BstBI (2240) Bpu10I (2248) KasI (2373) NarI (2374) SfoI (2375) PluTI (2377) PflMI (2609) NdeI (2617) ATG PpuMI (2667) SV40 NLS EagI - NotI (2701) BglII (2718) BspDI - ClaI (2842) BseRI (2987) BspQI - SapI (3180) pBridge™ 6527 bp
NsiI  (6241)
1 site
A T G C A T T A C G T A
PfoI  (5222)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
AatII  (5111)
1 site
G A C G T C C T G C A G
ZraI  (5109)
1 site
G A C G T C C T G C A G
ScaI  (4669)
1 site
A G T A C T T C A T G A
TsoI  (4588)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

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).
PvuI  (4559)
1 site
C G A T C G G C T A G C
BglI  (4309)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
BpmI  (4259)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

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.
AhdI  (4189)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
AlwNI  (3712)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AflIII  (3296)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (3296)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
PaeR7I  (651)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (651)
1 site
C T C G A G G A G C T C
HpaI  (711)
1 site
G T T A A C C A A T T G
BsrGI  (724)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
EcoRI  (878)
1 site
G A A T T C C T T A A G
TspMI  (883)
1 site
C C C G G G G G G C C C
XmaI  (883)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (885)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (888)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can
remain bound to DNA and alter its electrophoretic mobility.
SalI  (894)
1 site
G T C G A C C A G C T G
PstI  (904)
1 site
C T G C A G G A C G T C
PsiI  (963)
1 site
T T A T A A A A T A T T
NgoMIV  (1112)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (1114)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
BanII  (1331)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
AarI  (1398)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BstXI  (1541)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI sites may not be compatible.
Bsu36I  (1590)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
XbaI  (1747)
1 site
T C T A G A A G A T C T
BstAPI  (1759)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BstBI  (2240)
1 site
T T C G A A A A G C T T
Bpu10I  (2248)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
KasI  (2373)
1 site
G G C G C C C C G C G G
NarI  (2374)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (2375)
1 site
G G C G C C C C G C G G
PluTI  (2377)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
PflMI  (2609)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
NdeI  (2617)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
PpuMI  (2667)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
EagI  (2701)
1 site
C G G C C G G C C G G C
NotI  (2701)
1 site
G C G G C C G C C G C C G G C G
BglII  (2718)
1 site
A G A T C T T C T A G A
BspDI  (2842)
1 site
A T C G A T T A G C T A
ClaI  (2842)
1 site
A T C G A T T A G C T A
BseRI  (2987)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

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.
BspQI  (3180)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (3180)
1 site
G C T C T T C N C G A G A A G N N N N

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
   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
   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
MCS 1
878 .. 905  =  28 bp
multiple cloning site
MCS 1
878 .. 905  =  28 bp
multiple cloning site
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
MCS 2
2700 .. 2723  =  24 bp
multiple cloning site
MCS 2
2700 .. 2723  =  24 bp
multiple cloning site
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
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
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