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Plasmid Files

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

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
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

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.
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
   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
   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
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