pPICZ(alpha) A

Pichia pastoris vector for methanol-inducible expression of a secreted protein. For other reading frames, use pPICZα B or pPICZα C.

Sequence Author: Thermo Fisher (Invitrogen)

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BglII (1) AlwNI (3280) BssSI - BssSαI (3037) PciI (2864) MluI (2775) BsrGI (2768) EcoRV (2601) StuI * (2567) KflI (2559) DraIII (2516) FseI (2439) CsiI - SexAI * (2326) SgrAI (2277) SmaI (2253) TspMI - XmaI (2251) AatII (2209) ZraI (2207) BssHII - MauBI (2199) MscI (2166) NcoI - StyI (2161) AleI - MslI (76) BmrI (178) PflMI (190) Eco53kI (206) BanII - SacI (208) DraI - PmeI (413) BlpI (588) NsiI (677) BstXI - XcmI (706) MfeI (864) HindIII (872) BstBI (933) BstAPI (995) PsiI (1136) BsrDI (1155) EcoRI (1208) BsaAI - PmlI (1215) SfiI (1225) BsmBI - Esp3I (1236) Acc65I (1241) KpnI (1245) PspXI (1246) SacII (1255) NotI (1258) XbaI (1271) SalI (1314) AccI (1315) AgeI (1396) BsiWI (1528) PvuII (1562) BamHI (1677) BtgZI (1736) pPICZ α A 3593 bp
BglII  (1)
1 site
A G A T C T T C T A G A
AlwNI  (3280)
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.
BssSI  (3037)
1 site
C A C G A G G T G C T C
BssSαI  (3037)
1 site
C A C G A G G T G C T C
PciI  (2864)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
MluI  (2775)
1 site
A C G C G T T G C G C A
BsrGI  (2768)
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.
EcoRV  (2601)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
StuI  (2567)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
KflI  (2559)
1 site
G G G W C C C C C C W G G G

Sticky ends from different KflI sites may not be compatible.
DraIII  (2516)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
FseI  (2439)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
CsiI  (2326)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (2326)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
SgrAI  (2277)
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.
SmaI  (2253)
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  (2251)
1 site
C C C G G G G G G C C C
XmaI  (2251)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
AatII  (2209)
1 site
G A C G T C C T G C A G
ZraI  (2207)
1 site
G A C G T C C T G C A G
BssHII  (2199)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
MauBI  (2199)
1 site
C G C G C G C G G C G C G C G C
MscI  (2166)
1 site
T G G C C A A C C G G T
NcoI  (2161)
1 site
C C A T G G G G T A C C
StyI  (2161)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
AleI  (76)
1 site
C A C N N N N G T G G T G N N N N C A C
MslI  (76)
1 site
C A Y N N N N R T G G T R N N N N Y A C
BmrI  (178)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
PflMI  (190)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
Eco53kI  (206)
1 site
G A G C T C C T C G A G
BanII  (208)
1 site
G R G C Y C C Y C G