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Plasmid Files

pPIC9K

Pichia pastoris multi-copy integrating vector for methanol-inducible expression of a secreted protein.

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pPIC9K.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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AatII (9101) ZraI (9099) ScaI (8659) AflIII - PciI (7286) BspQI - SapI (7170) NdeI (7109) BstZ17I (7059) PflFI - Tth111I (7033) SphI (6036) SgrAI (5880) PspXI (5709) NruI (5654) SmaI (5437) TspMI - XmaI (5435) AsiSI (5313) BbvCI (4644) AleI (76) Eco53kI (206) SacI (208) PmeI (413) BlpI (588) XcmI (706) BamHI (938) PsiI (1144) SnaBI (1218) EcoRI (1222) AvrII (1228) EagI - NotI (1235) AgeI (1300) BtgZI (1754) FspAI (1826) XbaI (2032) Bsu36I (2088) HpaI (2138) SalI (3177) NcoI (3632) BspEI (3844) BsrGI (4298) pPIC9K 9276 bp
AatII  (9101)
1 site
G A C G T C C T G C A G
ZraI  (9099)
1 site
G A C G T C C T G C A G
ScaI  (8659)
1 site
A G T A C T T C A T G A
AflIII  (7286)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (7286)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (7170)
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  (7170)
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.
NdeI  (7109)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
BstZ17I  (7059)
1 site
G T A T A C C A T A T G
PflFI  (7033)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (7033)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
SphI  (6036)
1 site
G C A T G C C G T A C G
SgrAI  (5880)
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.
PspXI  (5709)
1 site
V C T C G A G B B G A G C T C V
NruI  (5654)
1 site
T C G C G A A G C G C T
SmaI  (5437)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (5435)
1 site
C C C G G G G G G C C C
XmaI  (5435)
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.
AsiSI  (5313)
1 site
G C G A T C G C C G C T A G C G
BbvCI  (4644)
1 site
C C T C A G C G G A G T C G
AleI  (76)
1 site
C A C N N N N G T G G T G N N N N C A C
Eco53kI  (206)
1 site
G A G C T C C T C G A G
SacI  (208)
1 site
G A G C T C C T C G A G
PmeI  (413)
1 site
G T T T A A A C C A A A T T T G
BlpI  (588)
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.
XcmI  (706)
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.
BamHI  (938)
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.
PsiI  (1144)
1 site
T T A T A A A A T A T T
SnaBI  (1218)
1 site
T A C G T A A T G C A T
EcoRI  (1222)
1 site
G A A T T C C T T A A G
AvrII  (1228)
1 site
C C T A G G G G A T C C
EagI  (1235)
1 site
C G G C C G G C C G G C
NotI  (1235)
1 site
G C G G C C G C C G C C G G C G
AgeI  (1300)
1 site
A C C G G T T G G C C A
BtgZI  (1754)
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.
FspAI  (1826)
1 site
R T G C G C A Y Y A C G C G T R
XbaI  (2032)
1 site
T C T A G A A G A T C T
Bsu36I  (2088)
1 site
C C T N A G G G G A