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Plasmid Files

pAUR112

Yeast centromeric vector with URA3 and aureobasidin A resistance markers.

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pAUR112.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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SalI (7091) XbaI (7085) BmgBI (6299) BsiWI (5582) BstEII (5576) EcoRI (5550) SacI (4760) Eco53kI (4758) KpnI (4754) Acc65I (4750) SmaI (4748) TspMI - XmaI (4746) Bpu10I (4597) BstBI (4132) PpuMI (4020) XcmI (3995) BsgI (3813) BglII (3408) AbsI - PaeR7I - PspXI - XhoI (7097) XmnI (444) FspI (821) BglI (926) BsrFI (959) AhdI (1044) BstAPI (2115) BsaAI (2182) PfoI (2287) SpeI (2388) NheI (3223) BmtI (3227) BtgZI (3335) BsrGI (3336) pAUR112 7104 bp
SalI  (7091)
1 site
G T C G A C C A G C T G
XbaI  (7085)
1 site
T C T A G A A G A T C T
BmgBI  (6299)
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.
BsiWI  (5582)
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.
BstEII  (5576)
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.
EcoRI  (5550)
1 site
G A A T T C C T T A A G
SacI  (4760)
1 site
G A G C T C C T C G A G
Eco53kI  (4758)
1 site
G A G C T C C T C G A G
KpnI  (4754)
1 site
G G T A C C C C A T G G
Acc65I  (4750)
1 site
G G T A C C C C A T G G
SmaI  (4748)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (4746)
1 site
C C C G G G G G G C C C
XmaI  (4746)
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.
Bpu10I  (4597)
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.
BstBI  (4132)
1 site
T T C G A A A A G C T T
PpuMI  (4020)
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.
XcmI  (3995)
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.
BsgI  (3813)
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).
BglII  (3408)
1 site
A G A T C T T C T A G A
AbsI  (7097)
1 site
C C T C G A G G G G A G C T C C
PaeR7I  (7097)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (7097)
1 site
V C T C G A G B B G A G C T C V
XhoI  (7097)
1 site
C T C G A G G A G C T C
XmnI  (444)
1 site
G A A N N N N T T C C T T N N N N A A G
FspI  (821)
1 site
T G C G C A A C G C G T
BglI  (926)
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  (959)
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  (1044)
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.
BstAPI  (2115)
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.
BsaAI  (2182)
1 site
Y A C G T R R T G C A Y
PfoI  (2287)
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.
SpeI  (2388)
1 site
A C T A G T T G A T C A
NheI  (3223)
1 site
G C T A G C C G A T C G
BmtI  (3227)
1 site
G C T A G C C G A T C G
BtgZI  (3335)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BsrGI  (3336)
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.
AurR
5128 .. 6333  =  1206 bp
401 amino acids  =  45.2 kDa
Product: aureobasidin A-resistant mutant of S. cerevisiae inositol phosphorylceramide synthase (Aeed et al., 2009)
confers fungal resistance to aureobasidin A
AurR
5128 .. 6333  =  1206 bp
401 amino acids  =  45.2 kDa
Product: aureobasidin A-resistant mutant of S. cerevisiae inositol phosphorylceramide synthase (Aeed et al., 2009)
confers fungal resistance to aureobasidin A
AmpR
257 .. 1117  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   257 .. 325  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
257 .. 1117  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   326 .. 1117  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
257 .. 1117  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
URA3
3867 .. 4670  =  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
3867 .. 4670  =  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
1288 .. 1876  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1288 .. 1876  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ARS1
3196 .. 3649  =  454 bp
S. cerevisiae autonomously replicating sequence ARS1
ARS1
3196 .. 3649  =  454 bp
S. cerevisiae autonomously replicating sequence ARS1
CEN4
2903 .. 3195  =  293 bp
S. cerevisiae CEN4 centromere
CEN4
2903 .. 3195  =  293 bp
S. cerevisiae CEN4 centromere
URA3 promoter
3651 .. 3866  =  216 bp
URA3 promoter
3651 .. 3866  =  216 bp
AmpR promoter
152 .. 256  =  105 bp
AmpR promoter
152 .. 256  =  105 bp
MCS
7085 .. 7103  =  19 bp
multiple cloning site
MCS
7085 .. 7103  =  19 bp
multiple cloning site
MCS
4746 .. 4761  =  16 bp
multiple cloning site
MCS
4746 .. 4761  =  16 bp
multiple cloning site
ORF:  5128 .. 6333  =  1206 bp
ORF:  401 amino acids  =  45.2 kDa
ORF:  257 .. 1117  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  3402 .. 3686  =  285 bp
ORF:  94 amino acids  =  10.8 kDa
ORF:  3867 .. 4670  =  804 bp
ORF:  267 amino acids  =  29.3 kDa
ORF:  5844 .. 6131  =  288 bp
ORF:  95 amino acids  =  11.5 kDa
ORF:  6858 .. 65  =  312 bp
ORF:  103 amino acids  =  12.4 kDa
ORF:  721 .. 987  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5343 .. 5567  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  6645 .. 7040  =  396 bp
ORF:  131 amino acids  =  15.0 kDa
ORF:  3902 .. 4378  =  477 bp
ORF:  158 amino acids  =  16.8 kDa
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