pPIC6 B

Pichia pastoris vector for methanol-inducible intracellular expression of a protein. For other reading frames, use pPIC6 A or pPIC6 C.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsYeast PlasmidspPIC6 B
BglII (1) AlwNI (3067) PciI (2651) MluI (2562) BsrGI (2555) BsrBI (2440) EcoRV (2388) StuI * (2354) KflI (2346) NmeAIII (2332) BmgBI (2327) DraIII (2303) TsoI (2265) PvuI (2133) NruI * (2129) BsgI - BtsI - BtsαI (2085) MscI (1901) NcoI - StyI (1896) AleI - MslI (76) PflMI (190) Eco53kI (206) SacI (208) DraI - PmeI (413) NsiI (677) BstXI - XcmI (706) MfeI (864) HindIII (872) BstBI (933) EcoRI (943) BsaAI - PmlI (950) NgoMIV (959) SfiI (960) NaeI (961) BsmBI - Esp3I (971) Acc65I (976) KpnI (980) AvaI - BsoBI - PaeR7I - PspXI - XhoI (981) SacII (990) NotI (993) XbaI (1006) SalI (1049) AccI (1050) AgeI (1131) BsiWI (1263) BamHI (1412) BtgZI (1471) pPIC6 B 3380 bp
BglII  (1)
1 site		 A G A T C T T C T A G A
AlwNI  (3067)
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  (2651)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
MluI  (2562)
1 site		 A C G C G T T G C G C A
BsrGI  (2555)
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  (2440)
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  (2388)
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  (2354)
1 site		 A G G C C T T C C G G A
* Blocked by Dcm methylation.
KflI  (2346)
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  (2332)
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  (2327)
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  (2303)
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  (2265)
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  (2133)
1 site		 C G A T C G G C T A G C
NruI  (2129)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
BsgI  (2085)
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  (2085)
1 site		 G C A G T G N N C G T C A C
BtsαI  (2085)
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  (1901)
1 site		 T G G C C A A C C G G T
NcoI  (1896)
1 site		 C C A T G G G G T A C C
StyI  (1896)
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
EcoRI  (943)
1 site		 G A A T T C C T T A A G
BsaAI  (950)
1 site		 Y A C G T R R T G C A Y
PmlI  (950)
1 site		 C A C G T G G T G C A C
NgoMIV  (959)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
SfiI  (960)
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.
NaeI  (961)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
BsmBI  (971)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (971)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
Acc65I  (976)
1 site		 G G T A C C C C A T G G
KpnI  (980)
1 site		 G G T A C C C C A T G G
AvaI  (981)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (981)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
PaeR7I  (981)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (981)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (981)
1 site		 C T C G A G G A G C T C
SacII  (990)
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  (993)
1 site		 G C G G C C G C C G C C G G C G
XbaI  (1006)
1 site		 T C T A G A A G A T C T
SalI  (1049)
1 site		 G T C G A C C A G C T G
AccI  (1050)
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  (1131)
1 site		 A C C G G T T G G C C A
BsiWI  (1263)
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.
BamHI  (1412)
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  (1471)
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.
AOX1 promoter
2 .. 940  =  939 bp
inducible promoter, regulated by methanol
AOX1 promoter
2 .. 940  =  939 bp
inducible promoter, regulated by methanol
ori
2712 .. 3300  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2712 .. 3300  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
TEF1 promoter
1413 .. 1824  =  412 bp
promoter for EF-1α
TEF1 promoter
1413 .. 1824  =  412 bp
promoter for EF-1α
BSD
1898 .. 2296  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
1898 .. 2296  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
CYC1 terminator
2390 .. 2637  =  248 bp
transcription terminator for CYC1
CYC1 terminator
2390 .. 2637  =  248 bp
transcription terminator for CYC1
AOX1 terminator
1152 .. 1398  =  247 bp
transcription terminator for AOX1
AOX1 terminator
1152 .. 1398  =  247 bp
transcription terminator for AOX1
Myc
1010 .. 1039  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
1010 .. 1039  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
6xHis
1055 .. 1072  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1055 .. 1072  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
EM7 promoter
1832 .. 1879  =  48 bp
synthetic bacterial promoter
EM7 promoter
1832 .. 1879  =  48 bp
synthetic bacterial promoter
MCS
932 .. 1011  =  80 bp
multiple cloning site
MCS
932 .. 1011  =  80 bp
multiple cloning site
ORF:  1898 .. 2296  =  399 bp
ORF:  132 amino acids  =  13.7 kDa
ORF:  486 .. 725  =  240 bp
ORF:  79 amino acids  =  9.2 kDa
ORF:  1844 .. 2200  =  357 bp
ORF:  118 amino acids  =  12.8 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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