pPIC6(alpha) B

Pichia pastoris vector for methanol-inducible expression of a secreted protein. For other reading frames, use pPIC6α A or pPIC6α C.

Sequence Author: Thermo Fisher (Invitrogen)

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BglII (1) AlwNI (3337) PciI (2921) MluI (2832) BsrGI (2825) BsrBI (2710) EcoRV (2658) StuI * (2624) KflI (2616) NmeAIII (2602) BmgBI (2597) DraIII (2573) TsoI (2535) PvuI (2403) NruI * (2399) BsgI - BtsI - BtsαI (2355) MscI (2171) NcoI - StyI (2166) AleI - MslI (76) PflMI (190) Eco53kI (206) SacI (208) DraI - PmeI (413) NsiI (677) BstXI - XcmI (706) MfeI (864) HindIII (872) BstBI (933) BstAPI (995) PsiI (1136) BsrDI (1155) PstI (1210) EcoRI (1212) BsaAI - PmlI (1219) NgoMIV (1228) SfiI (1229) NaeI (1230) BsmBI - Esp3I (1240) Acc65I (1245) KpnI (1249) PspXI (1250) SacII (1259) NotI (1262) XbaI (1275) SalI (1318) AccI (1319) AgeI (1400) BsiWI (1532) BamHI (1681) BtgZI (1740) pPIC6 α B 3650 bp
BglII  (1)
1 site
A G A T C T T C T A G A
AlwNI  (3337)
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.
PciI  (2921)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
MluI  (2832)
1 site
A C G C G T T G C G C A
BsrGI  (2825)
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.
BsrBI  (2710)
1 site
C C G C T C G G C G A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BsrBI will not always regenerate a BsrBI site.
BsrBI is typically used at 37°C, but can be used at temperatures up to 50°C.
EcoRV  (2658)
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  (2624)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
KflI  (2616)
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.
NmeAIII  (2602)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BmgBI  (2597)
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.
DraIII  (2573)
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.
TsoI  (2535)
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).
PvuI  (2403)
1 site
C G A T C G G C T A G C
NruI  (2399)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
BsgI  (2355)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BtsI  (2355)
1 site
G C A G T G N N C G T C A C
BtsαI  (2355)
1 site
G C A G T G N N C G T C A C

Sticky ends from different BtsαI sites may not be compatible.
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
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
SacI  (208)
1 site
G A G C T C C T C G A G
DraI  (413)
1 site
T T T A A A A A A T T T
PmeI  (413)
1 site
G T T T A A A C C A A A T T T G
NsiI  (677)
1 site
A T G C A T T A C G T A
BstXI  (706)
1 site
C C A N N N N N N T G G G G T N N N N N N A C C

Sticky ends from different BstXI 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.
MfeI  (864)
1 site
C A A T T G G T T A A C
HindIII  (872)
1 site
A A G C T T T T C G A A
BstBI  (933)
1 site
T T C G A A A A G C T T
BstAPI  (995)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
PsiI  (1136)
1 site
T T A T A A A A T A T T
BsrDI  (1155)
1 site
G C A A T G N N C