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Plasmid Files

pAUR224

Fission yeast (Schizosaccharomyces pombe) expression vector with an aureobasidin A resistance marker.

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pAUR224.dna
Map and Sequence File:    Download    Open   
Sequence Author:  TaKaRa
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NcoI (7400) SnaBI (7380) SwaI (6663) XcmI (6040) NsiI (5472) BbsI (5399) MscI (5231) BstAPI (5006) BsiWI (4963) PluTI (4452) SfoI (4450) NarI (4449) KasI (4448) NaeI (4251) NgoMIV (4249) DraIII (4148) PaeR7I - XhoI (1) Eco53kI (6) SacI (8) PstI (26) SalI (27) Acc65I (33) KpnI (37) SacII (40) PspOMI (41) TspMI - XmaI (44) ApaI (45) SmaI (46) BamHI (48) BclI * (70) HpaI (176) MluI (466) BlpI (767) AfeI (1168) BstZ17I (1282) AflII (1403) BspQI - SapI (1741) BseYI (2161) PspFI (2165) AlwNI (2273) AhdI (2750) BpmI (2820) NmeAIII (2898) EcoO109I (3726) pAUR224 7638 bp
NcoI  (7400)
1 site
C C A T G G G G T A C C
SnaBI  (7380)
1 site
T A C G T A A T G C A T
SwaI  (6663)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
XcmI  (6040)
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.
NsiI  (5472)
1 site
A T G C A T T A C G T A
BbsI  (5399)
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.
MscI  (5231)
1 site
T G G C C A A C C G G T
BstAPI  (5006)
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.
BsiWI  (4963)
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.
PluTI  (4452)
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  (4450)
1 site
G G C G C C C C G C G G
NarI  (4449)
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  (4448)
1 site
G G C G C C C C G C G G
NaeI  (4251)
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  (4249)
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.
DraIII  (4148)
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.
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1)
1 site
C T C G A G G A G C T C
Eco53kI  (6)
1 site
G A G C T C C T C G A G
SacI  (8)
1 site
G A G C T C C T C G A G
PstI  (26)
1 site
C T G C A G G A C G T C
SalI  (27)
1 site
G T C G A C C A G C T G
Acc65I  (33)
1 site
G G T A C C C C A T G G
KpnI  (37)
1 site
G G T A C C C C A T G G
SacII  (40)
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.
PspOMI  (41)
1 site
G G G C C C C C C G G G
TspMI  (44)
1 site
C C C G G G G G G C C C
XmaI  (44)
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.
ApaI  (45)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (46)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (48)
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.
BclI  (70)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
HpaI  (176)
1 site
G T T A A C C A A T T G
MluI  (466)
1 site
A C G C G T T G C G C A
BlpI  (767)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
AfeI  (1168)
1 site
A G C G C T T C G C G A
BstZ17I  (1282)
1 site
G T A T A C C A T A T G
AflII  (1403)
1 site
C T T A A G G A A T T C
BspQI  (1741)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1741)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
BseYI  (2161)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PspFI  (2165)
1 site
C C C A G C G G G T C G
AlwNI  (2273)
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  (2750)
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.
BpmI  (2820)
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.
NmeAIII  (2898)
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).
EcoO109I  (3726)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
aur1-R
5003 .. 6271  =  1269 bp
422 amino acids  =  47.5 kDa
Product: aureobasidin A resistance protein from Schizosaccharomyces pombe (Hashida-Okado et al., 1998)
aur1-R
5003 .. 6271  =  1269 bp
422 amino acids  =  47.5 kDa
Product: aureobasidin A resistance protein from Schizosaccharomyces pombe (Hashida-Okado et al., 1998)
AmpR
2677 .. 3537  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2677 .. 3468  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2677 .. 3537  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3469 .. 3537  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2677 .. 3537  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ars1
650 .. 1433  =  784 bp
Schizosaccharomyces pombe autonomously replicating sequence ars1
ars1
650 .. 1433  =  784 bp
Schizosaccharomyces pombe autonomously replicating sequence ars1
ori
1918 .. 2506  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1918 .. 2506  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
3924 .. 4379  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
3924 .. 4379  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
CMV enhancer
7101 .. 7404  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
7101 .. 7404  =  304 bp
human cytomegalovirus immediate early enhancer
CMV promoter
7405 .. 7608  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
7405 .. 7608  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
177 .. 298  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
177 .. 298  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3538 .. 3642  =  105 bp
AmpR promoter
3538 .. 3642  =  105 bp
MCS
1 .. 75  =  75 bp
multiple cloning site
MCS
1 .. 75  =  75 bp
multiple cloning site
ORF:  4399 .. 4785  =  387 bp
ORF:  128 amino acids  =  15.0 kDa
ORF:  2807 .. 3073  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  5003 .. 6271  =  1269 bp
ORF:  422 amino acids  =  47.5 kDa
ORF:  5649 .. 5894  =  246 bp
ORF:  81 amino acids  =  9.3 kDa
ORF:  2677 .. 3537  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  4948 .. 5289  =  342 bp
ORF:  113 amino acids  =  13.0 kDa
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