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pPICZ A

Pichia pastoris vector for methanol-inducible intracellular expression of a protein.

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pPICZ A.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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BglII (1) AlwNI (3016) BssS α I (2773) PciI (2600) MluI (2511) BsrGI (2504) EcoRV (2337) StuI * (2303) KflI (2295) DraIII (2252) FseI (2175) SexAI * (2062) SgrAI (2013) SmaI (1989) TspMI - XmaI (1987) AatII (1945) ZraI (1943) BssHII - MauBI (1935) MscI (1902) NcoI - StyI (1897) AleI - MslI (76) BmrI (178) PflMI (190) Eco53kI (206) SacI (208) DraI - PmeI (413) BlpI (588) NsiI (677) BstXI - XcmI (706) MfeI (864) HindIII (872) BstBI (933) EcoRI (943) BsaAI - PmlI (950) SfiI (960) BsmBI (971) Acc65I (976) KpnI (980) PaeR7I - PspXI - XhoI (981) SacII (990) NotI (993) PspOMI (1006) ApaI (1010) SalI (1051) AccI (1052) AgeI (1132) BsiWI (1264) PvuII (1298) BamHI (1413) BtgZI (1472) pPICZ A 3329 bp
BglII  (1)
1 site
A G A T C T T C T A G A
AlwNI  (3016)
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.
BssSαI  (2773)
1 site
C A C G A G G T G C T C
PciI  (2600)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
MluI  (2511)
1 site
A C G C G T T G C G C A
BsrGI  (2504)
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  (2337)
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  (2303)
1 site
A G G C C T T C C G G A
* Blocked by Dcm methylation.
KflI  (2295)
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  (2252)
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  (2175)
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.
SexAI  (2062)
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  (2013)
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  (1989)
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  (1987)
1 site
C C C G G G G G G C C C
XmaI  (1987)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
AatII  (1945)
1 site
G A C G T C C T G C A G
ZraI  (1943)
1 site
G A C G T C C T G C A G
BssHII  (1935)
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  (1935)
1 site
C G C G C G C G G C G C G C G C
MscI  (1902)
1 site
T G G C C A A C C G G T
NcoI  (1897)
1 site
C C A T G G G G T A C C
StyI  (1897)
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.
Unlike most restriction enzymes, BmrI can cleave DNA in the absence of magnesium.
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
BlpI  (588)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
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

PmlI gradually loses activity when stored at -20°C.
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.
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.
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
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
PspOMI  (1006)
1 site
G G G C C C C C C G G G
ApaI  (1010)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SalI  (1051)
1 site
G T C G A C C A G C T G
AccI  (1052)
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  (1132)
1 site
A C C G G T T G G C C A
BsiWI  (1264)
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.
PvuII  (1298)
1 site
C A G C T G G T C G A C
BamHI  (1413)
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  (1472)
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
2661 .. 3249  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2661 .. 3249  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
TEF1 promoter
1439 .. 1825  =  387 bp
promoter for EF-1α
TEF1 promoter
1439 .. 1825  =  387 bp
promoter for EF-1α
BleoR
1899 .. 2273  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
1899 .. 2273  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
CYC1 terminator
2339 .. 2586  =  248 bp
transcription terminator for CYC1
CYC1 terminator
2339 .. 2586  =  248 bp
transcription terminator for CYC1
AOX1 terminator
1153 .. 1399  =  247 bp
transcription terminator for AOX1
AOX1 terminator
1153 .. 1399  =  247 bp
transcription terminator for AOX1
EM7 promoter
1833 .. 1880  =  48 bp
synthetic bacterial promoter
EM7 promoter
1833 .. 1880  =  48 bp
synthetic bacterial promoter
Myc
1012 .. 1041  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
1012 .. 1041  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
6xHis
1057 .. 1074  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1057 .. 1074  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
MCS
932 .. 1011  =  80 bp
multiple cloning site
MCS
932 .. 1011  =  80 bp
multiple cloning site
ORF:  486 .. 725  =  240 bp
ORF:  79 amino acids  =  9.2 kDa
ORF:  1899 .. 2273  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  1845 .. 2342  =  498 bp
ORF:  165 amino acids  =  17.5 kDa
ORF:  1892 .. 2179  =  288 bp
ORF:  95 amino acids  =  10.1 kDa
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