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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.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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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) PpuMI (2667) 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