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Plasmid Files

pIZ/V5-His

Compact insect cell vector with a Zeocin™ resistance marker, for constitutive expression of proteins with an optional C-terminal V5-6xHis tag.

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pIZ V5-His.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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BsmI (2827) TsoI (2760) SalI (2736) PasI (2728) KflI - PpuMI (2724) DraIII (2662) SexAI * (2472) BmgBI (2443) SgrAI (2423) SmaI (2399) TspMI - XmaI (2397) AatII (2355) ZraI (2353) BssHII - MauBI (2345) MscI (2312) SfiI (2309) NcoI - StyI (2307) PmlI (2239) XmnI (2219) AcuI - Eco57MI (1493) HindIII (561) Acc65I (567) KpnI (571) Eco53kI (575) SacI (577) BamHI (579) SpeI (585) EcoRI (602) EcoRV (614) NotI (629) PaeR7I - PspXI - XhoI (635) XbaI (641) SacII (654) BsaI (696) MluI (702) AgeI (708) BspDI * - ClaI * (882) PciI (945) NspI (949) DrdI (1053) BciVI (1148) BseYI (1249) PspFI (1253) pIZ/V5-His 2876 bp
BsmI  (2827)
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.
TsoI  (2760)
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).
SalI  (2736)
1 site
G T C G A C C A G C T G
PasI  (2728)
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.
KflI  (2724)
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.
PpuMI  (2724)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
DraIII  (2662)
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.
SexAI  (2472)
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.
BmgBI  (2443)
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.
SgrAI  (2423)
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  (2399)
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  (2397)
1 site
C C C G G G G G G C C C
XmaI  (2397)
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  (2355)
1 site
G A C G T C C T G C A G
ZraI  (2353)
1 site
G A C G T C C T G C A G
BssHII  (2345)
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  (2345)
1 site
C G C G C G C G G C G C G C G C
MscI  (2312)
1 site
T G G C C A A C C G G T
SfiI  (2309)
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.
NcoI  (2307)
1 site
C C A T G G G G T A C C
StyI  (2307)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PmlI  (2239)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
XmnI  (2219)
1 site
G A A N N N N T T C C T T N N N N A A G
AcuI  (1493)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
Eco57MI  (1493)
1 site
C T G R A G ( N ) 14 N N G A C Y T C ( N ) 14

Sticky ends from different Eco57MI sites may not be compatible.
After cleavage, Eco57MI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
HindIII  (561)
1 site
A A G C T T T T C G A A
Acc65I  (567)
1 site
G G T A C C C C A T G G
KpnI  (571)
1 site
G G T A C C C C A T G G
Eco53kI  (575)
1 site
G A G C T C C T C G A G
SacI  (577)
1 site
G A G C T C C T C G A G
BamHI  (579)
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.
SpeI  (585)
1 site
A C T A G T T G A T C A
EcoRI  (602)
1 site
G A A T T C C T T A A G
EcoRV  (614)
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.
NotI  (629)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (635)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (635)
1 site
V C T C G A G B B G A G C T C V
XhoI  (635)
1 site
C T C G A G G A G C T C
XbaI  (641)
1 site
T C T A G A A G A T C T
SacII  (654)
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.
BsaI  (696)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
MluI  (702)
1 site
A C G C G T T G C G C A
AgeI  (708)
1 site
A C C G G T T G G C C A
BspDI  (882)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (882)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
PciI  (945)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (949)
1 site
R C A T G Y Y G T A C R
DrdI  (1053)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
BciVI  (1148)
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.
BseYI  (1249)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PspFI  (1253)
1 site
C C C A G C G G G T C G
ori
1006 .. 1594  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
1006 .. 1594  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
OpIE-2 promoter
5 .. 552  =  548 bp
strong constitutive baculovirus promoter for insect cell expression
OpIE-2 promoter
5 .. 552  =  548 bp
strong constitutive baculovirus promoter for insect cell expression
OpIE-2 promoter
1666 .. 2213  =  548 bp
strong constitutive baculovirus promoter for insect cell expression
OpIE-2 promoter
1666 .. 2213  =  548 bp
strong constitutive baculovirus promoter for insect cell expression
BleoR
2309 .. 2683  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
BleoR
2309 .. 2683  =  375 bp
124 amino acids  =  13.8 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and Zeocin™
SV40 poly(A) signal
2747 .. 4  =  134 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2747 .. 4  =  134 bp
SV40 polyadenylation signal
OpIE-2 poly(A) signal
749 .. 878  =  130 bp
baculovirus polyadenylation signal
OpIE-2 poly(A) signal
749 .. 878  =  130 bp
baculovirus polyadenylation signal
MCS
561 .. 656  =  96 bp
multiple cloning site
MCS
561 .. 656  =  96 bp
multiple cloning site
EM7 promoter
2242 .. 2289  =  48 bp
synthetic bacterial promoter
EM7 promoter
2242 .. 2289  =  48 bp
synthetic bacterial promoter
V5 tag
663 .. 704  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
V5 tag
663 .. 704  =  42 bp
14 amino acids  =  1.4 kDa
Product: epitope tag from simian virus 5
6xHis
714 .. 731  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
714 .. 731  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
ORF:  19 .. 249  =  231 bp
ORF:  76 amino acids  =  8.9 kDa
ORF:  2309 .. 2683  =  375 bp
ORF:  124 amino acids  =  13.8 kDa
ORF:  1680 .. 1910  =  231 bp
ORF:  76 amino acids  =  8.9 kDa
ORF:  2293 .. 2589  =  297 bp
ORF:  98 amino acids  =  10.4 kDa
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