pPICZ(alpha) C

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

Sequence Author: Thermo Fisher (Invitrogen)

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BglII (1) AlwNI (3285) BssSI - BssSαI (3042) PciI (2869) MluI (2780) BsrGI (2773) EcoRV (2606) StuI * (2572) KflI (2564) DraIII (2521) FseI (2444) CsiI - SexAI * (2331) SgrAI (2282) SmaI (2258) TspMI - XmaI (2256) AatII (2214) ZraI (2212) BssHII - MauBI (2204) MscI (2171) NcoI - StyI (2166) 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) BspDI - ClaI (1208) EcoRI (1213) BsaAI - PmlI (1220) SfiI (1230) BsmBI - Esp3I (1241) Acc65I (1246) KpnI (1250) PspXI (1251) SacII (1260) NotI (1263) XbaI (1276) SalI (1319) AccI (1320) AgeI (1401) BsiWI (1533) PvuII (1567) BamHI (1682) BtgZI (1741) pPICZ α C 3598 bp
BglII  (1)
1 site
A G A T C T T C T A G A
AlwNI  (3285)
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  (3042)
1 site
C A C G A G G T G C T C
BssSαI  (3042)
1 site
C A C G A G G T G C T C
PciI  (2869)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
MluI  (2780)
1 site
A C G C G T T G C G C A
BsrGI  (2773)
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  (2606)
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  (2572)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
KflI  (2564)
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  (2521)
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  (2444)
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  (2331)
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  (2331)
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  (2282)
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  (2258)
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  (2256)
1 site
C C C G G G G G G C C C
XmaI  (2256)
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  (2214)
1 site
G A C G T C C T G C A G
ZraI  (2212)
1 site
G A C G T C C T G C A G
BssHII  (2204)
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  (2204)
1 site
C G C G C G C G G C G C G C G C
MscI  (2171)
1 site
T G G C C A A C C G G T
NcoI  (2166)
1 site
C C A T G G G G T A C C
StyI  (2166)
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.