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pPINKα-HC

Pichia pastoris integrating vector for high-copy expression of a secreted protein in the PichiaPink™ system.

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pPINK(alpha)-HC.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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PluTI (7736) SfoI (7734) NarI (7733) KasI (7732) NdeI (7681) AatII (7432) ZraI (7430) AhdI (6510) AflII (5007) PshAI (4968) BmgBI (4689) SpeI (4554) EcoNI (4422) BspEI (4101) BglII (1) Eco53kI (206) SacI (208) PmeI (413) XcmI (706) AvaI - BsoBI - PaeR7I - XhoI (1184) BmeT110I (1185) StuI (1195) Bsu36I (1197) Acc65I (1201) KpnI (1205) NgoMIV (1208) NaeI (1210) FseI (1212) SwaI (1218) EcoRV (1246) BsrGI (1415) MluI (1422) BamHI (1503) NotI - SacII (1510) NheI (1593) BmtI (1597) BbsI (2262) DraIII (2547) BsaAI (3263) BsgI (3370) AscI - BssHII (3423) NruI (3509) BsaBI (3614) pPINK α- HC 7898 bp
PluTI  (7736)
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  (7734)
1 site
G G C G C C C C G C G G
NarI  (7733)
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  (7732)
1 site
G G C G C C C C G C G G
NdeI  (7681)
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.
AatII  (7432)
1 site
G A C G T C C T G C A G
ZraI  (7430)
1 site
G A C G T C C T G C A G
AhdI  (6510)
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.
AflII  (5007)
1 site
C T T A A G G A A T T C
PshAI  (4968)
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.
BmgBI  (4689)
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.
SpeI  (4554)
1 site
A C T A G T T G A T C A
EcoNI  (4422)
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.
BspEI  (4101)
1 site
T C C G G A A G G C C T
BglII  (1)
1 site
A G A T C T T C T A G A
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
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.
AvaI  (1184)
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  (1184)
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.
PaeR7I  (1184)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1184)
1 site
C T C G A G G A G C T C
BmeT110I  (1185)
1 site
C Y C G R G G R G C Y C
StuI  (1195)
1 site
A G G C C T T C C G G A
Bsu36I  (1197)
1 site
C C T N A G G G G A N T C