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Plasmid Files

pAUR123

Yeast centromeric vector with an aureobasidin A resistance marker, for expressing a gene under control of the ADH1 promoter and terminator.

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pAUR123.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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pAUR123 R Primer (6701 .. 6724) HpaI (6621) SacI - XbaI (6612) Eco53kI (6610) PaeR7I - PspXI - XhoI (6605) SalI (6600) SmaI (6597) KpnI - TspMI - XmaI (6595) Acc65I (6591) pAUR123 F Primer (6550 .. 6568) BsgI (6536) DraIII (5205) BmgBI (5188) EcoRV (5069) NsiI (4672) ApaI (4496) PspOMI (4492) BsiWI (4471) BstEII (4465) EcoRI (4439) HindIII (3993) SbfI (3441) XmnI (444) FspI (821) BglI (926) AhdI (1044) NdeI (2111) BstAPI (2115) BsaAI (2182) PfoI (2287) BsmBI (2289) SpeI (2388) NheI (3223) BmtI (3227) BtgZI (3335) BsrGI (3336) BglII (3408) pAUR123 6982 bp
HpaI  (6621)
1 site
G T T A A C C A A T T G
SacI  (6612)
1 site
G A G C T C C T C G A G
XbaI  (6612)
1 site
T C T A G A A G A T C T
Eco53kI  (6610)
1 site
G A G C T C C T C G A G
PaeR7I  (6605)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (6605)
1 site
V C T C G A G B B G A G C T C V
XhoI  (6605)
1 site
C T C G A G G A G C T C
SalI  (6600)
1 site
G T C G A C C A G C T G
SmaI  (6597)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
KpnI  (6595)
1 site
G G T A C C C C A T G G
TspMI  (6595)
1 site
C C C G G G G G G C C C
XmaI  (6595)
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.
Acc65I  (6591)
1 site
G G T A C C C C A T G G
BsgI  (6536)
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).
DraIII  (5205)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
BmgBI  (5188)
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.
EcoRV  (5069)
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.
NsiI  (4672)
1 site
A T G C A T T A C G T A
ApaI  (4496)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (4492)
1 site
G G G C C C C C C G G G
BsiWI  (4471)
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  (4465)
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  (4439)
1 site
G A A T T C C T T A A G
HindIII  (3993)
1 site
A A G C T T T T C G A A
SbfI  (3441)
1 site
C C T G C A G G G G A C G T C 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.
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.
NdeI  (2111)
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.
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.
BsmBI  (2289)
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.
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.
BglII  (3408)
1 site
A G A T C T T C T A G A
pAUR123 R Primer
24-mer  /  33% GC
1 binding site
6701 .. 6724  =  24 annealed bases
Tm  =  53°C
pAUR123 F Primer
19-mer  /  37% GC
1 binding site
6550 .. 6568  =  19 annealed bases
Tm  =  50°C
AurR
4017 .. 5222  =  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
4017 .. 5222  =  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
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
ADH1 promoter
6195 .. 6590  =  396 bp
promoter for alcohol dehydrogenase 1
ADH1 promoter
6195 .. 6590  =  396 bp
promoter for alcohol dehydrogenase 1
CEN4
2903 .. 3195  =  293 bp
S. cerevisiae CEN4 centromere
CEN4
2903 .. 3195  =  293 bp
S. cerevisiae CEN4 centromere
ADH1 terminator
6628 .. 6815  =  188 bp
transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene
ADH1 terminator
6628 .. 6815  =  188 bp
transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene
AmpR promoter
152 .. 256  =  105 bp
AmpR promoter
152 .. 256  =  105 bp
MCS
6591 .. 6624  =  34 bp
multiple cloning site
MCS
6591 .. 6624  =  34 bp
multiple cloning site
ORF:  257 .. 1117  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  4733 .. 5020  =  288 bp
ORF:  95 amino acids  =  11.5 kDa
ORF:  5747 .. 5986  =  240 bp
ORF:  79 amino acids  =  9.2 kDa
ORF:  6782 .. 48  =  249 bp
ORF:  82 amino acids  =  9.1 kDa
ORF:  3402 .. 3755  =  354 bp
ORF:  117 amino acids  =  13.4 kDa
ORF:  4017 .. 5222  =  1206 bp
ORF:  401 amino acids  =  45.2 kDa
ORF:  4232 .. 4456  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  5534 .. 5929  =  396 bp
ORF:  131 amino acids  =  15.0 kDa
ORF:  721 .. 987  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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