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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
ORF:  2302 .. 3321  =  1020 bp
ORF:  339 amino acids  =  37.8 kDa
ORF:  3997 .. 4269  =  273 bp
ORF:  90 amino acids  =  10.0 kDa
ORF:  4719 .. 5579  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  87 .. 659  =  573 bp
ORF:  190 amino acids  =  21.2 kDa
ORF:  3120 .. 3365  =  246 bp
ORF:  81 amino acids  =  8.6 kDa
ORF:  4052 .. 4333  =  282 bp
ORF:  93 amino acids  =  10.8 kDa
ORF:  5183 .. 5449  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  64 .. 294  =  231 bp
ORF:  76 amino acids  =  8.6 kDa
ORF:  1714 .. 1953  =  240 bp
ORF:  79 amino acids  =  8.9 kDa
ORF:  2236 .. 3054  =  819 bp
ORF:  272 amino acids  =  30.3 kDa
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