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Plasmid Files

pIZT/V5-His

Insect cell vector with a GFP-Zeocin™ resistance fusion gene, for constitutive expression of proteins with an optional C-terminal V5-6xHis tag.

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pIZT V5-His Sequence and MappIZT V5-His.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 BsmI (3287) DraIII (3122) BglI (3074) SexAI * (2932) BmgBI (2903) SgrAI (2883) SmaI (2859) TspMI - XmaI (2857) AatII (2815) ZraI (2813) MauBI (2805) BtgZI (2650) MfeI (2629) BsrGI (2345) NdeI (2299) BspEI * (2291) MscI (2240) BpmI (2116) BmtI (2074) NheI (2070) AseI (1994) PmlI (1983) BlpI (1697) BspHI * (4) BsiWI (321) MslI (461) KasI (478) NarI (479) SfoI (480) PluTI (482) Acc65I (567) KpnI (571) Eco53kI (575) SacI (577) SpeI (585) EcoRI (602) EcoRV (614) NotI (629) XbaI (641) EcoO109I (647) SacII (654) BsaI (696) AgeI (708) BspDI * - ClaI * (882) PciI (945) BciVI (1148) pIZT/V5-His 3336 bp
BsmI  (3287)
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.
DraIII  (3122)
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.
BglI  (3074)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
SexAI  (2932)
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  (2903)
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  (2883)
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  (2859)
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  (2857)
1 site
C C C G G G G G G C C C
XmaI  (2857)
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  (2815)
1 site
G A C G T C C T G C A G
ZraI  (2813)
1 site
G A C G T C C T G C A G
MauBI  (2805)
1 site
C G C G C G C G G C G C G C G C
BtgZI  (2650)
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.
MfeI  (2629)
1 site
C A A T T G G T T A A C
BsrGI  (2345)
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.
NdeI  (2299)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
BspEI  (2291)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
MscI  (2240)
1 site
T G G C C A A C C G G T
BpmI  (2116)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI
recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its
electrophoretic mobility.
BpmI quickly loses activity at 37°C.
BmtI  (2074)
1 site
G C T A G C C G A T C G
NheI  (2070)
1 site
G C T A G C C G A T C G
AseI  (1994)
1 site
A T T A A T T A A T T A
PmlI  (1983)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BlpI  (1697)
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.
BspHI  (4)
1 site
T C A T G A A G T A C T
* Blocked by Dam methylation.
BsiWI  (321)
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.
MslI  (461)
1 site
C A Y N N N N R T G G T R N N N N Y A C
KasI  (478)
1 site
G G C G C C C C G C G G
NarI  (479)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (480)
1 site
G G C G C C C C G C G G
PluTI  (482)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
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
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
XbaI  (641)
1 site
T C T A G A A G A T C T
EcoO109I  (647)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
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.
AgeI  (708)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
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.
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.
Cycle 3 GFP
2067 .. 2765  =  699 bp
233 amino acids  =  26.1 kDa
Product: Cycle 3 GFP (Crameri et al., 1996) lacking
the unstructured C-terminal peptide
mammalian codon-optimized
Cycle 3 GFP
2067 .. 2765  =  699 bp
233 amino acids  =  26.1 kDa
Product: Cycle 3 GFP (Crameri et al., 1996) lacking
the unstructured C-terminal peptide
mammalian codon-optimized
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
BleoR
2766 .. 3143  =  378 bp
125 amino acids  =  13.9 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
BleoR
2766 .. 3143  =  378 bp
125 amino acids  =  13.9 kDa
Product: antibiotic-binding protein
confers resistance to bleomycin, phleomycin, and
Zeocin™
OpIE-1 promoter
1669 .. 1960  =  292 bp
moderate constitutive baculovirus promoter for
insect cell expression
OpIE-1 promoter
1669 .. 1960  =  292 bp
moderate constitutive baculovirus promoter for
insect cell expression
SV40 poly(A) signal
3207 .. 4  =  134 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3207 .. 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
567 .. 656  =  90 bp
multiple cloning site
MCS
567 .. 656  =  90 bp
multiple cloning site
EM7 promoter
1986 .. 2033  =  48 bp
synthetic bacterial promoter
EM7 promoter
1986 .. 2033  =  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
ATG
2052 .. 2054  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
2052 .. 2054  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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