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Plasmid Files

pAUR101

Yeast chromosomal integrating vector with an aureobasidin A resistance marker.

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pAUR101.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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ZraI (67) PfoI (6636) BsaAI (6276) NaeI (6173) NgoMIV (6171) PluTI (5976) SfoI (5974) NarI (5973) KasI (5972) SacI (5809) Eco53kI (5807) KpnI (5803) Acc65I (5799) SmaI (5797) TspMI - XmaI (5795) XbaI (5784) SalI (5778) SbfI (5776) SphI (5770) BbsI (5648) BmgBI (4976) BstZ17I (4514) NsiI (4460) ApaI (4284) PspOMI (4280) BsiWI (4259) BstEII (4253) StuI (4143) AatII (69) XmnI (388) NmeAIII (841) BpmI (919) AhdI (988) BseRI (3022) MscI (3231) pAUR101 6687 bp
ZraI  (67)
1 site
G A C G T C C T G C A G
PfoI  (6636)
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.
BsaAI  (6276)
1 site
Y A C G T R R T G C A Y
NaeI  (6173)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (6171)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
PluTI  (5976)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (5974)
1 site
G G C G C C C C G C G G
NarI  (5973)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (5972)
1 site
G G C G C C C C G C G G
SacI  (5809)
1 site
G A G C T C C T C G A G
Eco53kI  (5807)
1 site
G A G C T C C T C G A G
KpnI  (5803)
1 site
G G T A C C C C A T G G
Acc65I  (5799)
1 site
G G T A C C C C A T G G
SmaI  (5797)
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  (5795)
1 site
C C C G G G G G G C C C
XmaI  (5795)
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.
XbaI  (5784)
1 site
T C T A G A A G A T C T
SalI  (5778)
1 site
G T C G A C C A G C T G
SbfI  (5776)
1 site
C C T G C A G G G G A C G T C C
SphI  (5770)
1 site
G C A T G C C G T A C G
BbsI  (5648)
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.
BmgBI  (4976)
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.
BstZ17I  (4514)
1 site
G T A T A C C A T A T G
NsiI  (4460)
1 site
A T G C A T T A C G T A
ApaI  (4284)
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  (4280)
1 site
G G G C C C C C C G G G
BsiWI  (4259)
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  (4253)
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.
StuI  (4143)
1 site
A G G C C T T C C G G A
AatII  (69)
1 site
G A C G T C C T G C A G
XmnI  (388)
1 site
G A A N N N N T T C C T T N N N N A A G
NmeAIII  (841)
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).
BpmI  (919)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AhdI  (988)
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.
BseRI  (3022)
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  (3231)
1 site
T G G C C A A C C G G T
AurR
3805 .. 5010  =  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
3805 .. 5010  =  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
201 .. 1061  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   201 .. 269  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
201 .. 1061  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   270 .. 1061  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
201 .. 1061  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
1232 .. 1820  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1232 .. 1820  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6046 .. 6501  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6046 .. 6501  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
96 .. 200  =  105 bp
AmpR promoter
96 .. 200  =  105 bp
MCS
5766 .. 5810  =  45 bp
multiple cloning site
MCS
5766 .. 5810  =  45 bp
multiple cloning site
M13 fwd
5817 .. 5833  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
5817 .. 5833  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  3805 .. 5010  =  1206 bp
ORF:  401 amino acids  =  45.2 kDa
ORF:  201 .. 1061  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4521 .. 4808  =  288 bp
ORF:  95 amino acids  =  11.5 kDa
ORF:  5535 .. 5843  =  309 bp
ORF:  102 amino acids  =  11.8 kDa
ORF:  2152 .. 3102  =  951 bp
ORF:  316 amino acids  =  36.4 kDa
ORF:  2145 .. 2384  =  240 bp
ORF:  79 amino acids  =  9.2 kDa
ORF:  4020 .. 4244  =  225 bp
ORF:  74 amino acids  =  8.1 kDa
ORF:  5322 .. 5717  =  396 bp
ORF:  131 amino acids  =  15.0 kDa
ORF:  5742 .. 6026  =  285 bp
ORF:  94 amino acids  =  11.0 kDa
ORF:  665 .. 931  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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