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Plasmid Files

pEF1α-Tet3G

Vector encoding the Tet-On® 3G transactivator protein, which allows for inducible gene expression.

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pEF1alpha-Tet3G Sequence and MappEF1alpha-Tet3G.dna
Map and Sequence File   
Sequence Author:  Clontech
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 BstZ17I (7809) AccI (7808) SfiI (7355) AvrII (7301) BmtI (7287) NheI (7283) BclI * (6956) EagI (6891) MscI (6719) RsrII (6282) BstBI (6117) Bsu36I (5922) BspEI * (5791) PflMI (5643) PspXI (4929) AatII (4859) ZraI (4857) ScaI (4417) PvuI (4307) BsaI (3998) BsrGI (47) AleI (182) AgeI (283) BstXI (516) AflII (588) FseI (913) BbvCI (1111) DraIII (1245) EcoRI (1486) XbaI (1498) PmlI (1626) NdeI (2074) PpuMI (2535) BstEII (2631) HindIII (2718) AhdI (3937) pEF1 α- Tet3G 7859 bp
BstZ17I  (7809)
1 site
G T A T A C C A T A T G
AccI  (7808)
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.
SfiI  (7355)
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.
AvrII  (7301)
1 site
C C T A G G G G A T C C
BmtI  (7287)
1 site
G C T A G C C G A T C G
NheI  (7283)
1 site
G C T A G C C G A T C G
BclI  (6956)
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.
EagI  (6891)
1 site
C G G C C G G C C G G C
MscI  (6719)
1 site
T G G C C A A C C G G T
RsrII  (6282)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII
recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BstBI  (6117)
1 site
T T C G A A A A G C T T
Bsu36I  (5922)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
BspEI  (5791)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
PflMI  (5643)
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.
PspXI  (4929)
1 site
V C T C G A G B B G A G C T C V
AatII  (4859)
1 site
G A C G T C C T G C A G
ZraI  (4857)
1 site
G A C G T C C T G C A G
ScaI  (4417)
1 site
A G T A C T T C A T G A
PvuI  (4307)
1 site
C G A T C G G C T A G C
BsaI  (3998)
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.
BsrGI  (47)
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.
AleI  (182)
1 site
C A C N N N N G T G G T G N N N N C A C
AgeI  (283)
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.
BstXI  (516)
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.
AflII  (588)
1 site
C T T A A G G A A T T C

The sticky ends produced by AflII are hard to ligate.
FseI  (913)
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.
BbvCI  (1111)
1 site
C C T C A G C G G A G T C G
DraIII  (1245)
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.
EcoRI  (1486)
1 site
G A A T T C C T T A A G
XbaI  (1498)
1 site
T C T A G A A G A T C T
PmlI  (1626)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
NdeI  (2074)
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.
PpuMI  (2535)
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.
BstEII  (2631)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
HindIII  (2718)
1 site
A A G C T T T T C G A A
AhdI  (3937)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
EF-1α promoter
206 .. 1387  =  1182 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
206 .. 1387  =  1182 bp
strong constitutive promoter for human elongation
factor EF-1α
AmpR
3864 .. 4724  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   3864 .. 4655  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3864 .. 4724  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   4656 .. 4724  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3864 .. 4724  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
6136 .. 6930  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
6136 .. 6930  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
Tet-On® 3G
1495 .. 2241  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
Tet-On® 3G
1495 .. 2241  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
ori
3105 .. 3693  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
3105 .. 3693  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
SV40 promoter
7291 .. 7620  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
7291 .. 7620  =  330 bp
SV40 enhancer and early promoter
SV40 poly(A) signal
2251 .. 2385  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2251 .. 2385  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4942 .. 5076  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
4942 .. 5076  =  135 bp
SV40 polyadenylation signal
AmpR promoter
4725 .. 4829  =  105 bp
AmpR promoter
4725 .. 4829  =  105 bp
small t intron
5651 .. 5716  =  66 bp
simian virus 40 (SV40) small t antigen intron
small t intron
5651 .. 5716  =  66 bp
simian virus 40 (SV40) small t antigen intron
EF-1α intron A
436 .. 1378  =  943 bp
intron upstream of the start codon
EF-1α intron A
436 .. 1378  =  943 bp
intron upstream of the start codon
SV40 ori
7305 .. 7440  =  136 bp
SV40 origin of replication
SV40 ori
7305 .. 7440  =  136 bp
SV40 origin of replication
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