pGBDU-C1

Yeast two-hybrid "bait" vector with a URA3 marker for fusing a gene to the GAL4 DNA binding domain. For other reading frames, use pGBDU-C2 or pGBDU-C3.
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BsaAI - SnaBI (5355) BmgBI (4913) PfoI (4692) AatII (4581) ZraI (4579) TsoI (4058) PvuI (4029) NmeAIII (3807) BglI (3779) AhdI (3659) BanI (3607) PaeR7I - XhoI (651) HpaI (711) BsrGI (724) EcoRI (878) TspMI - XmaI (884) SmaI (886) BamHI (890) BspDI - ClaI (897) SalI (902) PstI (912) BglII (914) BseRI (1020) MscI (1050) BstXI (1182) NdeI (1626) PpuMI (1862) EcoRV (1897) NcoI (1914) BsmI (2071) PspOMI * (2083) ApaI * (2087) StuI (2145) Bpu10I (2439) pGBDU-C1 5997 bp
BsaAI  (5355)
1 site
Y A C G T R R T G C A Y
SnaBI  (5355)
1 site
T A C G T A A T G C A T
BmgBI  (4913)
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  (4692)
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  (4581)
1 site
G A C G T C C T G C A G
ZraI  (4579)
1 site
G A C G T C C T G C A G
TsoI  (4058)
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  (4029)
1 site
C G A T C G G C T A G C
NmeAIII  (3807)
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  (3779)
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.
AhdI  (3659)
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.
BanI  (3607)
1 site
G G Y R C C C C R Y G G

Sticky ends from different BanI sites may not be compatible.
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  (884)
1 site
C C C G G G G G G C C C
XmaI  (884)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (886)
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  (890)
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  (897)
1 site
A T C G A T T A G C T A
ClaI  (897)
1 site
A T C G A T T A G C T A
SalI  (902)
1 site
G T C G A C C A G C T G
PstI  (912)
1 site
C T G C A G G A C G T C
BglII  (914)
1 site
A G A T C T T C T A G A
BseRI  (1020)
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  (1050)
1 site
T G G C C A A C C G G T
BstXI  (1182)
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.
NdeI  (1626)
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  (1862)
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.
EcoRV  (1897)
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.
NcoI  (1914)
1 site
C C A T G G G G T A C C
BsmI  (2071)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
PspOMI  (2083)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI  (2087)
1 site
G G G C C C C C C G G G
* Blocked by Dcm methylation.
ApaI can be used between 25°C and 37°C.
StuI  (2145)
1 site
A G G C C T T C C G G A
Bpu10I  (2439)
1 site
C C T N A G C G G A N T C G

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.
2μ ori
4833 .. 5997  =  1165 bp
yeast 2μ plasmid origin of replication
2μ ori
4833 .. 5997  =  1165 bp
yeast 2μ plasmid origin of replication
AmpR
3586 .. 4446  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   3586 .. 4377  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3586 .. 4446  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4378 .. 4446  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
3586 .. 4446  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
URA3
1709 .. 2512  =  804 bp
267 amino acids  =  29.3 kDa
Product: orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis
yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)
URA3
1709 .. 2512  =  804 bp
267 amino acids  =  29.3 kDa
Product: orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis
yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)
ori
2827 .. 3415  =  589 bp
high-copy-number colE1/pMB1/pBR322/pUC origin of replication
ori
2827 .. 3415  =  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
URA3 promoter
1492 .. 1708  =  217 bp
URA3 promoter
1492 .. 1708  =  217 bp
ADH1 terminator
1294 .. 1481  =  188 bp
transcription terminator for alcohol dehydrogenase 1
ADH1 terminator
1294 .. 1481  =  188 bp
transcription terminator for alcohol dehydrogenase 1
AmpR promoter
4447 .. 4551  =  105 bp
AmpR promoter
4447 .. 4551  =  105 bp
MCS
878 .. 919  =  42 bp
multiple cloning site
MCS
878 .. 919  =  42 bp
multiple cloning site
ORF:  4930 .. 5196  =  267 bp
ORF:  88 amino acids  =  10.3 kDa
ORF:  434 .. 937  =  504 bp
ORF:  167 amino acids  =  19.2 kDa
ORF:  1709 .. 2512  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  3716 .. 3982  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1744 .. 2220  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
ORF:  3586 .. 4446  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1517 .. 1747  =  231 bp
ORF:  76 amino acids  =  8.9 kDa
ORF:  5672 .. 5962  =  291 bp
ORF:  96 amino acids  =  11.5 kDa
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Individual Sequences & Maps

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