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Plasmid Files

pUCATPH

Hygromycin resistance plasmid for restriction enzyme-mediated integration (REMI) mutagenesis of fungi.

 
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 SspI (4903) AhdI (4099) AlwNI (3622) PciI (3206) BspQI - SapI (3090) lac operator EcoRI (2850) SacI (2848) Eco53kI (2846) KpnI (2842) Acc65I (2838) SmaI (2836) BmeT110I (2835) AvaI - BsoBI - TspMI - XmaI (2834) BamHI (2829) XbaI (2823) SalI (2817) PfoI (46) HindIII (399) SphI (409) PstI - SbfI (415) SalI (417) EcoNI (445) SphI (510) AfeI (576) SgrAI (658) BamHI (693) XbaI (699) BstZ17I (836) PasI (939) BseRI (1002) BlpI (1121) MluI (1225) BamHI (1411) SacII (1677) BtgZI (1779) RsrII (2044) AsiSI (2093) PstI (2122) EcoRI (2208) PshAI (2428) BspDI - ClaI (2458) AflII (2519) pUCATPH 5086 bp
SspI  (4903)
1 site
A A T A T T T T A T A A
AhdI  (4099)
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.
AlwNI  (3622)
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.
PciI  (3206)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3090)
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  (3090)
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.
EcoRI  (2850)
2 sites
G A A T T C C T T A A G
SacI  (2848)
1 site
G A G C T C C T C G A G
Eco53kI  (2846)
1 site
G A G C T C C T C G A G
KpnI  (2842)
1 site
G G T A C C C C A T G G
Acc65I  (2838)
1 site
G G T A C C C C A T G G
SmaI  (2836)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BmeT110I  (2835)
1 site
C Y C G R G G R G C Y C
AvaI  (2834)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2834)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures
up to 65°C.
TspMI  (2834)
1 site
C C C G G G G G G C C C
XmaI  (2834)
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.
BamHI  (2829)
3 sites
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.
XbaI  (2823)
2 sites
T C T A G A A G A T C T
SalI  (2817)
2 sites
G T C G A C C A G C T G
PfoI  (46)
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.
HindIII  (399)
1 site
A A G C T T T T C G A A
SphI  (409)
2 sites
G C A T G C C G T A C G
PstI  (415)
2 sites
C T G C A G G A C G T C
SbfI  (415)
1 site
C C T G C A G G G G A C G T C C
SalI  (417)
2 sites
G T C G A C C A G C T G
EcoNI  (445)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
SphI  (510)
2 sites
G C A T G C C G T A C G
AfeI  (576)
1 site
A G C G C T T C G C G A
SgrAI  (658)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
BamHI  (693)
3 sites
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.
XbaI  (699)
2 sites
T C T A G A A G A T C T
BstZ17I  (836)
1 site
G T A T A C C A T A T G
PasI  (939)
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.
BseRI  (1002)
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.
BlpI  (1121)
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  (1225)
1 site
A C G C G T T G C G C A
BamHI  (1411)
3 sites
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.
SacII  (1677)
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.
BtgZI  (1779)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
RsrII  (2044)
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.
AsiSI  (2093)
1 site
G C G A T C G C C G C T A G C G
PstI  (2122)
2 sites
C T G C A G G A C G T C
EcoRI  (2208)
2 sites
G A A T T C C T T A A G
PshAI  (2428)
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.
BspDI  (2458)
1 site
A T C G A T T A G C T A
ClaI  (2458)
1 site
A T C G A T T A G C T A
AflII  (2519)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
HygR
1431 .. 2456  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
HygR
1431 .. 2456  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E.
coli

confers resistance to hygromycin
AmpR
4026 .. 4886  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   4026 .. 4817  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4026 .. 4886  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4818 .. 4886  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
4026 .. 4886  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
3267 .. 3855  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3267 .. 3855  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
trpC terminator
704 .. 1262  =  559 bp
terminator for Aspergillus nidulans trpC
trpC terminator
704 .. 1262  =  559 bp
terminator for Aspergillus nidulans trpC
trpC promoter
2461 .. 2817  =  357 bp
promoter for Aspergillus nidulans trpC
trpC promoter
2461 .. 2817  =  357 bp
promoter for Aspergillus nidulans trpC
AmpR promoter
4887 .. 4991  =  105 bp
AmpR promoter
4887 .. 4991  =  105 bp
lac promoter
2913 .. 2943  =  31 bp
   Segment 3:  -10  
   2913 .. 2919  =  7 bp
promoter for the E. coli lac operon
lac promoter
2913 .. 2943  =  31 bp
   Segment 2:  
   2920 .. 2937  =  18 bp
promoter for the E. coli lac operon
lac promoter
2913 .. 2943  =  31 bp
   Segment 1:  -35  
   2938 .. 2943  =  6 bp
promoter for the E. coli lac operon
lac promoter
2913 .. 2943  =  31 bp
3 segments
promoter for the E. coli lac operon
M13 fwd
379 .. 395  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 fwd
379 .. 395  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2865 .. 2881  =  17 bp
common sequencing primer, one of multiple similar
variants
M13 rev
2865 .. 2881  =  17 bp
common sequencing primer, one of multiple similar
variants
lac operator
2889 .. 2905  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
2889 .. 2905  =  17 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
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