pGAPZ(alpha) C

Pichia pastoris vector for constitutive expression of a secreted protein. For other reading frames, use pGAPZα A or pGAPZα B.

Sequence Author: Thermo Fisher (Invitrogen)

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NsiI (3145) AlwNI (2835) BciVI (2622) BssSI - BssSαI (2592) PciI (2419) MluI (2330) BsrGI (2323) AclI (2289) EcoRV (2156) StuI * (2122) KflI (2114) DraIII (2071) FseI (1994) CsiI - SexAI * (1881) SgrAI (1832) SmaI (1808) TspMI - XmaI (1806) AatII (1764) ZraI (1762) BssHII - MauBI (1754) MscI (1721) NcoI (1716) AseI (1660) BglII (1) TsoI (61) BfuAI - BspMI (96) AvrII (190) BspQI - SapI (251) PasI (301) PshAI (351) BspHI (355) DraI (439) MfeI (469) BstBI (485) BstAPI (547) PsiI (688) BspDI - ClaI (760) EcoRI (765) BsaAI - PmlI (772) SfiI (782) Acc65I (798) KpnI (802) PspXI (803) SacII (812) NotI (815) XbaI (828) SalI (871) AccI (872) AgeI (952) BsmI (993) BsiWI (1084) BclI * (1100) PvuII (1118) BamHI (1233) BtgZI (1292) pGAPZ α C 3152 bp
NsiI  (3145)
1 site
A T G C A T T A C G T A
AlwNI  (2835)
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.
BciVI  (2622)
1 site
G T A T C C ( N ) 5 N C A T A G G ( N ) 5

The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
BssSI  (2592)
1 site
C A C G A G G T G C T C
BssSαI  (2592)
1 site
C A C G A G G T G C T C
PciI  (2419)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
MluI  (2330)
1 site
A C G C G T T G C G C A
BsrGI  (2323)
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.
AclI  (2289)
1 site
A A C G T T T T G C A A
EcoRV  (2156)
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  (2122)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
KflI  (2114)
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  (2071)
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  (1994)
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  (1881)
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  (1881)
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  (1832)
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  (1808)
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  (1806)
1 site
C C C G G G G G G C C C
XmaI  (1806)
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  (1764)
1 site
G A C G T C C T G C A G
ZraI  (1762)
1 site
G A C G T C C T G C A G
BssHII  (1754)
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  (1754)
1 site
C G C G C G C G G C G C G C G C
MscI  (1721)
1 site
T G G C C A A C C G G T
NcoI  (1716)
1 site
C C A T G G G G T A C C
AseI  (1660)
1 site
A T T A A T T A A T T A
BglII  (1)
1 site
A G A T C T T C T A G A
TsoI  (61)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
BfuAI  (96)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (96)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
AvrII  (190)
1 site
C C