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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HomePlasmidsYeast PlasmidspGAPZ(alpha) C
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 T A G G G G A T C C
BspQI  (251)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (251)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PasI  (301)
1 site		 C C C W G G G G G G W C C C
Sticky ends from different PasI sites may not be compatible.
PshAI  (351)
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.
BspHI  (355)
1 site		 T C A T G A A G T A C T
DraI  (439)
1 site		 T T T A A A A A A T T T
MfeI  (469)
1 site		 C A A T T G G T T A A C
BstBI  (485)
1 site		 T T C G A A A A G C T T
BstAPI  (547)
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  (688)
1 site		 T T A T A A A A T A T T
BspDI  (760)
1 site		 A T C G A T T A G C T A
ClaI  (760)
1 site		 A T C G A T T A G C T A
EcoRI  (765)
1 site		 G A A T T C C T T A A G
BsaAI  (772)
1 site		 Y A C G T R R T G C A Y
PmlI  (772)
1 site		 C A C G T G G T G C A C
SfiI  (782)
1 site		 G G C C N N N N N G G C C C C G G N N N N N C C G G
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
Acc65I  (798)
1 site		 G G T A C C C C A T G G
KpnI  (802)
1 site		 G G T A C C C C A T G G
PspXI  (803)
1 site		 V C T C G A G B B G A G C T C V
SacII  (812)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
NotI  (815)
1 site		 G C G G C C G C C G C C G G C G
XbaI  (828)
1 site		 T C T A G A A G A T C T
SalI  (871)
1 site		 G T C G A C C A G C T G
AccI  (872)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
AgeI  (952)
1 site		 A C C G G T T G G C C A
BsmI  (993)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
BsiWI  (1084)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
BclI  (1100)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
PvuII  (1118)
1 site		 C A G C T G G T C G A C
BamHI  (1233)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BtgZI  (1292)
1 site		 G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4
Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
ori
2480 .. 3068  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2480 .. 3068  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
GAP promoter
7 .. 483  =  477 bp
promoter for Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase
GAP promoter
7 .. 483  =  477 bp
promoter for Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase
TEF1 promoter
1234 .. 1645  =  412 bp
promoter for EF-1α
TEF1 promoter
1234 .. 1645  =  412 bp
promoter for EF-1α
α-factor secretion signal
493 .. 759  =  267 bp
89 amino acids  =  9.3 kDa
3 segments
   Segment 1:  presequence  
   493 .. 549  =  57 bp
   19 amino acids  =  2.0 kDa
Product: N-terminal secretion signal from S. cerevisiae alpha-factor
Cleavage by the Kex2 protease occurs after the dibasic KR sequence. The EA dipeptides are then removed by dipeptidyl aminopeptidase A.
α-factor secretion signal
493 .. 759  =  267 bp
89 amino acids  =  9.3 kDa
3 segments
   Segment 2:  pro region  
   550 .. 747  =  198 bp
   66 amino acids  =  7.0 kDa
Product: N-terminal secretion signal from S. cerevisiae alpha-factor
Cleavage by the Kex2 protease occurs after the dibasic KR sequence. The EA dipeptides are then removed by dipeptidyl aminopeptidase A.
α-factor secretion signal
493 .. 759  =  267 bp
89 amino acids  =  9.3 kDa
3 segments
   Segment 3:  
   748 .. 759  =  12 bp
   4 amino acids  =  418.4 Da
Product: N-terminal secretion signal from S. cerevisiae alpha-factor
Cleavage by the Kex2 protease occurs after the dibasic KR sequence. The EA dipeptides are then removed by dipeptidyl aminopeptidase A.
α-factor secretion signal
493 .. 759  =  267 bp
89 amino acids  =  9.3 kDa
3 segments
Product: N-terminal secretion signal from S. cerevisiae alpha-factor
Cleavage by the Kex2 protease occurs after the dibasic KR sequence. The EA dipeptides are then removed by dipeptidyl aminopeptidase A.
Myc
832 .. 861  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
832 .. 861  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
6xHis
877 .. 894  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
877 .. 894  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
BleoR
1718 .. 2092  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
1718 .. 2092  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
CYC1 terminator
2158 .. 2405  =  248 bp
transcription terminator for CYC1
CYC1 terminator
2158 .. 2405  =  248 bp
transcription terminator for CYC1
AOX1 terminator
973 .. 1219  =  247 bp
transcription terminator for AOX1
AOX1 terminator
973 .. 1219  =  247 bp
transcription terminator for AOX1
EM7 promoter
1652 .. 1699  =  48 bp
synthetic bacterial promoter
EM7 promoter
1652 .. 1699  =  48 bp
synthetic bacterial promoter
MCS
759 .. 833  =  75 bp
multiple cloning site
MCS
759 .. 833  =  75 bp
multiple cloning site
ORF:  136 .. 360  =  225 bp
ORF:  74 amino acids  =  8.0 kDa
ORF:  493 .. 897  =  405 bp
ORF:  134 amino acids  =  14.2 kDa
ORF:  1718 .. 2092  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  612 .. 845  =  234 bp
ORF:  77 amino acids  =  8.2 kDa
ORF:  1664 .. 2161  =  498 bp
ORF:  165 amino acids  =  17.5 kDa
ORF:  1711 .. 1998  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
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Individual Sequences & Maps

bleMX6
pFA6a-kanMX6
pPIC6(alpha) B
pYES2.1-E
hphMX6
pFA6a-kanMX6-PGAL1
pPIC6(alpha) C
pYES2 CT
kanMX
pFA6a-kanMX6-PGAL1-3HA
pPIC9
pYES2 NT A
kanMX2
pFA6a-kanMX6-PGAL1-GFP
pPIC9K
pYES2 NT B
kanMX4
pFA6a-kanMX6-PGAL1-GST
pPICZ A
pYES2 NT C
kanMX6
pFA6a-natMX6
pPICZ B
pYES3 CT
natMX6
pFA6a-TRP1
pPICZ C
pYES6 CT
pAbAi
pFA6a-TRP1-PGAL1
pPICZ(alpha) A
pYM25
pAD-GAL4
pFA6a-TRP1-PGAL1-3HA
pPICZ(alpha) B
pYM26
pAG25
pFA6a-TRP1-PGAL1-GFP
pPICZ(alpha) C
pYM44
pAG29
pFA6a-TRP1-PGAL1-GST
pPink-HC
YCp50
pAG32
pFLD
pPink-LC
YCp50-poly
pAO815
pFLD(alpha)
pPINK(alpha)-HC
YCplac111
patMX4
pGAD-C1
pRS303
YCplac22
pAUR101
pGAD-C2
pRS304
YCplac33
pAUR112
pGAD-C3
pRS305
YEp13
pAUR123
pGADT7 AD
pRS306
YEp24
pAUR224
pGADT7-Rec
pRS313
YEp351
pBridge
pGADT7-Rec (linearized)
pRS314
YEp352
pDH25
pGAPZ A
pRS315
YEp353
pEG202
pGAPZ B
pRS316
YEp354
pESC-HIS
pGAPZ C
pRS403
YEp355
pESC-LEU
pGAPZ(alpha) A
pRS404
YEp356
pESC-TRP
pGAPZ(alpha) B
pRS405
YEp356R
pESC-URA
pGAPZ(alpha) C
pRS406
YEp357
pFA6-kanMX4
pGBD-C1
pRS413
YEp357R
pFA6a
pGBD-C2
pRS414
YEp358
pFA6a-12Pk-kanMX6
pGBD-C3
pRS415
YEp358R
pFA6a-13Myc-His3MX6
pGBDU-C1
pRS416
YEp363
pFA6a-13Myc-kanMX6
pGBDU-C2
pRS423
YEp364
pFA6a-13Myc-TRP1
pGBDU-C3
pRS424
YEp365
pFA6a-3HA-His3MX6
pGBKT7
pRS425
YEp366
pFA6a-3HA-kanMX6
pGilda
pRS426
YEp366R
pFA6a-3HA-TRP1
pHIL-D2
pSpot5
YEp367
pFA6a-5FLAG-kanMX6
pHIL-S1
pSpot6
YEp367R
pFA6a-bleMX6
pJG4-5
pSpot7
YEp368
pFA6a-GFP(S65T)-His3MX6
pKLAC1
pSpot8
YEp368R
pFA6a-GFP(S65T)-kanMX6
pKLAC2
pTEF1 Zeo
YEplac112
pFA6a-GFP(S65T)-TRP1
pOAD
pUCATPH
YEplac181
pFA6a-GST-His3MX6
pOBD2
pY-secSUMOstar
YEplac195
pFA6a-GST-kanMX6
pP-secSUMO3
pY-SUMOstar
YIp351
pFA6a-GST-TRP1
pPIC3
pYC2-E
YIp352
pFA6a-His3MX6
pPIC3.5
pYC2 CT
YIp5
pFA6a-His3MX6-PGAL1
pPIC3.5K
pYC2 NT A
YIplac128
pFA6a-His3MX6-PGAL1-3HA
pPIC6 A
pYC2 NT B
YIplac204
pFA6a-His3MX6-PGAL1-GFP
pPIC6 B
pYC2 NT C
YIplac211
pFA6a-His3MX6-PGAL1-GST
pPIC6 C
pYC6 CT
YRp7
pFA6a-hphMX6
pPIC6(alpha) A
pYES2
alpha-factor secretion signal

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