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Plasmid Files

pAbAi

Yeast integrating vector for cloning a DNA bait sequence to be used in a one-hybrid screen.

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pAbAi Sequence and MappAbAi.dna
Map and Sequence File   
Sequence Author:  Clontech
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 HindIII (1) NdeI (4762) BsgI (4733) SbfI (4699) BstBI (4410) BbsI (4399) BsmBI (4051) Bpu10I (3944) PsiI (3862) BsaHI (3416) PvuI (3249) FspI (3101) BglI (2999) BsrFI (2959) AhdI (2879) Eco53kI (15) SacI (17) Acc65I (19) KpnI - TspMI - XmaI (23) SmaI (25) SalI (34) AbsI - PaeR7I - PspXI - XhoI (40) BamHI (338) BstEII (633) BsiWI (639) BmgBI (1356) AflII (1386) PvuII (1810) DrdI (2094) pAbAi 4894 bp
HindIII  (1)
1 site
A A G C T T T T C G A A
NdeI  (4762)
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.
BsgI  (4733)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
SbfI  (4699)
1 site
C C T G C A G G G G A C G T C C
BstBI  (4410)
1 site
T T C G A A A A G C T T
BbsI  (4399)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BsmBI  (4051)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
Bpu10I  (3944)
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.
PsiI  (3862)
1 site
T T A T A A A A T A T T
BsaHI  (3416)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
PvuI  (3249)
1 site
C G A T C G G C T A G C
FspI  (3101)
1 site
T G C G C A A C G C G T
BglI  (2999)
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.
BsrFI  (2959)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
AhdI  (2879)
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.
Eco53kI  (15)
1 site
G A G C T C C T C G A G
SacI  (17)
1 site
G A G C T C C T C G A G
Acc65I  (19)
1 site
G G T A C C C C A T G G
KpnI  (23)
1 site
G G T A C C C C A T G G
TspMI  (23)
1 site
C C C G G G G G G C C C
XmaI  (23)
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  (25)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SalI  (34)
1 site
G T C G A C C A G C T G
AbsI  (40)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (40)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (40)
1 site
V C T C G A G B B G A G C T C V
XhoI  (40)
1 site
C T C G A G G A G C T C
BamHI  (338)
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.
BstEII  (633)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
BsiWI  (639)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
BmgBI  (1356)
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.
AflII  (1386)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
PvuII  (1810)
1 site
C A G C T G G T C G A C
DrdI  (2094)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
AurR
185 .. 1390  =  1206 bp
401 amino acids  =  45.2 kDa
Product: aureobasidin A-resistant mutant of inositol
phosphorylceramide synthase
confers fungal resistance to Aureobasidin A
AurR
185 .. 1390  =  1206 bp
401 amino acids  =  45.2 kDa
Product: aureobasidin A-resistant mutant of inositol
phosphorylceramide synthase
confers fungal resistance to Aureobasidin A
AmpR
2806 .. 3666  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2806 .. 3597  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2806 .. 3666  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3598 .. 3666  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2806 .. 3666  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
URA3
3875 .. 4678  =  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
3875 .. 4678  =  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
2047 .. 2635  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2047 .. 2635  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
URA3 promoter
4679 .. 4894  =  216 bp
URA3 promoter
4679 .. 4894  =  216 bp
AmpR promoter
3667 .. 3771  =  105 bp
AmpR promoter
3667 .. 3771  =  105 bp
MCS
1 .. 45  =  45 bp
multiple cloning site
MCS
1 .. 45  =  45 bp
multiple cloning site
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