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Plasmid Files

pGBD-C3

Yeast two-hybrid "bait" vector for fusing a gene to the GAL4 DNA binding domain. For other reading frames, use pGBD‑C1 or pGBD‑C2.

 
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 NsiI (5610) SnaBI (5254) BmgBI (4812) PfoI (4591) EcoO109I (4534) AatII (4480) ZraI (4478) ScaI (4038) TsoI (3957) PvuI (3928) NmeAIII (3706) BglI (3678) BpmI (3628) AhdI (3558) AlwNI (3081) BstBI (166) PaeR7I - XhoI (651) HpaI (711) BsrGI (724) EcoRI (878) TspMI - XmaI (883) SmaI (885) BamHI (889) BspDI - ClaI (896) SalI (901) PstI (911) BglII (913) BseRI (1019) MscI (1049) XcmI (1106) PsiI (1335) NgoMIV (1484) NaeI (1486) BanII (1703) AarI (1770) EcoRV (1922) Bsu36I (1962) MfeI (2063) XbaI (2119) BstAPI (2131) PmlI (2247) PvuII (2489) BspQI - SapI (2549) AflIII - PciI (2665) pGBD-C3 5896 bp
NsiI  (5610)
1 site
A T G C A T T A C G T A
SnaBI  (5254)
1 site
T A C G T A A T G C A T
BmgBI  (4812)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends
generated by BmgBI will not always regenerate a BmgBI site.
PfoI  (4591)
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.
EcoO109I  (4534)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
AatII  (4480)
1 site
G A C G T C C T G C A G
ZraI  (4478)
1 site
G A C G T C C T G C A G
ScaI  (4038)
1 site
A G T A C T T C A T G A
TsoI  (3957)
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  (3928)
1 site
C G A T C G G C T A G C
NmeAIII  (3706)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BglI  (3678)
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  (3628)
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  (3558)
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  (3081)
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.
BstBI  (166)
1 site
T T C G A A A A G C T T
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  (889)
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.
BspDI  (896)
1 site
A T C G A T T A G C T A
ClaI  (896)
1 site
A T C G A T T A G C T A
SalI  (901)
1 site
G T C G A C C A G C T G
PstI  (911)
1 site
C T G C A G G A C G T C
BglII  (913)
1 site
A G A T C T T C T A G A
BseRI  (1019)
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.
MscI  (1049)
1 site
T G G C C A A C C G G T
XcmI  (1106)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
PsiI  (1335)
1 site
T T A T A A A A T A T T
NgoMIV  (1484)
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  (1486)
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  (1703)
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  (1770)
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.
EcoRV  (1922)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to
delete a base after cleavage.
Bsu36I  (1962)
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.
MfeI  (2063)
1 site
C A A T T G G T T A A C
XbaI  (2119)
1 site
T C T A G A A G A T C T
BstAPI  (2131)
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.
PmlI  (2247)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
PvuII  (2489)
1 site
C A G C T G G T C G A C
BspQI  (2549)
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  (2549)
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.
AflIII  (2665)
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  (2665)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
2μ ori
4732 .. 5896  =  1165 bp
yeast 2μ plasmid origin of replication
2μ ori
4732 .. 5896  =  1165 bp
yeast 2μ plasmid origin of replication
AmpR
3485 .. 4345  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3485 .. 4276  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3485 .. 4345  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4277 .. 4345  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3485 .. 4345  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
TRP1
1533 .. 2207  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase,
required for tryptophan biosynthesis
yeast auxotrophic marker
TRP1
1533 .. 2207  =  675 bp
224 amino acids  =  24.1 kDa
Product: phosphoribosylanthranilate isomerase,
required for tryptophan biosynthesis
yeast auxotrophic marker
ori
2726 .. 3314  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin
of replication
ori
2726 .. 3314  =  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
5 .. 406  =  402 bp
promoter for alcohol dehydrogenase 1
ADH1 promoter
5 .. 406  =  402 bp
promoter for alcohol dehydrogenase 1
ADH1 terminator
1293 .. 1480  =  188 bp
transcription terminator for alcohol dehydrogenase 1
ADH1 terminator
1293 .. 1480  =  188 bp
transcription terminator for alcohol dehydrogenase 1
AmpR promoter
4346 .. 4450  =  105 bp
AmpR promoter
4346 .. 4450  =  105 bp
TRP1 promoter
2208 .. 2309  =  102 bp
TRP1 promoter
2208 .. 2309  =  102 bp
MCS
878 .. 918  =  41 bp
multiple cloning site
MCS
878 .. 918  =  41 bp
multiple cloning site
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