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Plasmid Files

pAUR316

Autonomously replicating E. coli-Aspergillus shuttle vector with an aureobasidin A resistance marker.

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pAUR316.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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PciI (11,202) SbfI (10,943) BglII (10,932) AflII (10,928) AfeI (10,926) BamHI (10,917) SacI (8145) Eco53kI (8143) AlwNI (416) AhdI (893) BmrI (933) PshAI (1919) PspXI (2039) Acc65I (2211) KpnI (2215) BsaAI - PmlI (2878) NcoI (2928) ZraI (3034) AatII (3036) SrfI (3155) BstXI (3351) BssHII (3560) Bsu36I (4129) SexAI * (4701) MluI (5108) SacII (5512) pAUR316 11,662 bp
PciI  (11,202)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
SbfI  (10,943)
1 site
C C T G C A G G G G A C G T C C
BglII  (10,932)
1 site
A G A T C T T C T A G A
AflII  (10,928)
1 site
C T T A A G G A A T T C
AfeI  (10,926)
1 site
A G C G C T T C G C G A
BamHI  (10,917)
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.
SacI  (8145)
1 site
G A G C T C C T C G A G
Eco53kI  (8143)
1 site
G A G C T C C T C G A G
AlwNI  (416)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (893)
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.
BmrI  (933)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
PshAI  (1919)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
PspXI  (2039)
1 site
V C T C G A G B B G A G C T C V
Acc65I  (2211)
1 site
G G T A C C C C A T G G
KpnI  (2215)
1 site
G G T A C C C C A T G G
BsaAI  (2878)
1 site
Y A C G T R R T G C A Y
PmlI  (2878)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
NcoI  (2928)
1 site
C C A T G G G G T A C C
ZraI  (3034)
1 site
G A C G T C C T G C A G
AatII  (3036)
1 site
G A C G T C C T G C A G
SrfI  (3155)
1 site
G C C C G G G C C G G G C C C G
BstXI  (3351)
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.
BssHII  (3560)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
Bsu36I  (4129)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
SexAI  (4701)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
MluI  (5108)
1 site
A C G C G T T G C G C A
SacII  (5512)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
AMA1
5682 .. 10,908  =  5227 bp
replication origin from Aspergillus nidulans (Aleksenko and Clutterbuck, 1996)
AMA1
5682 .. 10,908  =  5227 bp
replication origin from Aspergillus nidulans (Aleksenko and Clutterbuck, 1996)
aurA-R
2859 .. 4234  =  1376 bp
439 amino acids  =  49.6 kDa
2 segments
Product: aureobasidin A resistance protein from Aspergillus nidulans (Kuroda et al., 1999)
aurA-R
2859 .. 4234  =  1376 bp
439 amino acids  =  49.6 kDa
   Segment 1:  
   2859 .. 3280  =  422 bp
   141 amino acids  =  16.3 kDa
Product: aureobasidin A resistance protein from Aspergillus nidulans (Kuroda et al., 1999)
aurA-R
2859 .. 4234  =  1376 bp
439 amino acids  =  49.6 kDa
   Segment 2:  
   3337 .. 4234  =  898 bp
   298 amino acids  =  33.3 kDa
Product: aureobasidin A resistance protein from Aspergillus nidulans (Kuroda et al., 1999)
aurA-R
2859 .. 4234  =  1376 bp
439 amino acids  =  49.6 kDa
2 segments
Product: aureobasidin A resistance protein from Aspergillus nidulans (Kuroda et al., 1999)
AmpR
820 .. 1680  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   820 .. 1611  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
820 .. 1680  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   1612 .. 1680  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
820 .. 1680  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
61 .. 649  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
61 .. 649  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
1681 .. 1785  =  105 bp
AmpR promoter
1681 .. 1785  =  105 bp
MCS
10,917 .. 10,945  =  29 bp
multiple cloning site
MCS
10,917 .. 10,945  =  29 bp
multiple cloning site
ORF:  11,293 .. 386  =  756 bp
ORF:  251 amino acids  =  28.0 kDa
ORF:  950 .. 1216  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3521 .. 4234  =  714 bp
ORF:  237 amino acids  =  26.5 kDa
ORF:  2859 .. 3284  =  426 bp
ORF:  141 amino acids  =  16.3 kDa
ORF:  3903 .. 4142  =  240 bp
ORF:  79 amino acids  =  8.6 kDa
ORF:  11,520 .. 91  =  234 bp
ORF:  77 amino acids  =  8.9 kDa
ORF:  1577 .. 1888  =  312 bp
ORF:  103 amino acids  =  12.1 kDa
ORF:  820 .. 1680  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  8146 .. 8397  =  252 bp
ORF:  83 amino acids  =  9.2 kDa
ORF:  2715 .. 2960  =  246 bp
ORF:  81 amino acids  =  9.2 kDa
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