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Plasmid Files

pAN7-1

Vector with a hygromycin B resistance marker for transforming Aspergillus species.

 
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pAN7-1.dna
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 PciI (6394) AhdI (5506) PluTI (4283) SfoI (4281) NarI (4280) KasI (4279) HindIII (4115) SbfI (4107) XbaI (4051) NaeI (3970) NgoMIV (3968) BstZ17I (3918) PasI (3812) BlpI (3630) MluI (3525) BglII (138) BmgBI (374) BstEII (396) StuI (465) AarI (828) BclI * (853) NheI (904) BmtI (908) PspXI (1076) XcmI (1163) BsrGI (1422) AvrII (1718) MfeI (1734) SalI (1944) BbvCI (1976) PshAI (2323) AsiSI (2660) RsrII (2704) BamHI (3339) pAN7-1 6749 bp
PciI  (6394)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (5506)
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.
PluTI  (4283)
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  (4281)
1 site
G G C G C C C C G C G G
NarI  (4280)
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  (4279)
1 site
G G C G C C C C G C G G
HindIII  (4115)
1 site
A A G C T T T T C G A A
SbfI  (4107)
1 site
C C T G C A G G G G A C G T C C
XbaI  (4051)
1 site
T C T A G A A G A T C T
NaeI  (3970)
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  (3968)
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.
BstZ17I  (3918)
1 site
G T A T A C C A T A T G
PasI  (3812)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
BlpI  (3630)
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.
MluI  (3525)
1 site
A C G C G T T G C G C A
BglII  (138)
1 site
A G A T C T T C T A G A
BmgBI  (374)
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.
BstEII  (396)
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  (465)
1 site
A G G C C T T C C G G A
AarI  (828)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BclI  (853)
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.
NheI  (904)
1 site
G C T A G C C G A T C G
BmtI  (908)
1 site
G C T A G C C G A T C G
PspXI  (1076)
1 site
V C T C G A G B B G A G C T C V
XcmI  (1163)
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.
BsrGI  (1422)
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.
AvrII  (1718)
1 site
C C T A G G G G A T C C
MfeI  (1734)
1 site
C A A T T G G T T A A C
SalI  (1944)
1 site
G T C G A C C A G C T G
BbvCI  (1976)
1 site
C C T C A G C G G A G T C G
PshAI  (2323)
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.
AsiSI  (2660)
1 site
G C G A T C G C C G C T A G C G
RsrII  (2704)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BamHI  (3339)
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.
gpdA promoter
1 .. 2129  =  2129 bp
promoter from the Aspergillus nidulans
glyceraldehyde-3-phosphate dehydrogenase gene
(Punt et al., 1990)
gpdA promoter
1 .. 2129  =  2129 bp
promoter from the Aspergillus nidulans
glyceraldehyde-3-phosphate dehydrogenase gene
(Punt et al., 1990)
HygR
2302 .. 3321  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
2302 .. 3321  =  1020 bp
339 amino acids  =  37.8 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
AmpR
4719 .. 5579  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4719 .. 4787  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4719 .. 5579  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4788 .. 5579  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4719 .. 5579  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
trpC terminator
3339 .. 4101  =  763 bp
transcription terminator from the Aspergillus
nidulans trpC
gene (Punt et al., 1987)
trpC terminator
3339 .. 4101  =  763 bp
transcription terminator from the Aspergillus
nidulans trpC
gene (Punt et al., 1987)
ori
5750 .. 6338  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5750 .. 6338  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
gpdA intron
2178 .. 2293  =  116 bp
intron from the Aspergillus nidulans
glyceraldehyde-3-phosphate dehydrogenase gene
(Punt et al., 1990)
gpdA intron
2178 .. 2293  =  116 bp
intron from the Aspergillus nidulans
glyceraldehyde-3-phosphate dehydrogenase gene
(Punt et al., 1990)
AmpR promoter
4614 .. 4718  =  105 bp
AmpR promoter
4614 .. 4718  =  105 bp
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