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Plasmid Files

pLVX-EF1α-Tet3G

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

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pLVX-EF1alpha-Tet3G Sequence and MappLVX-EF1alpha-Tet3G.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SgrDI (9712) SspI (9595) PvuI (9161) AhdI (8791) BmtI (7517) NheI (7513) PfoI (6275) MluI (5989) AfeI (5928) RsrII (5655) PflFI - Tth111I (5257) MscI (5221) KflI - PpuMI (1934) BsrGI (2229) AgeI (2465) BsmBI (3595) NdeI (4249) TspMI - XmaI (4408) SmaI (4410) BamHI (4417) AvrII (4576) AarI (4764) BmgBI (4967) pLVX-EF1 α- Tet3G 10,080 bp
SgrDI  (9712)
1 site
C G T C G A C G G C A G C T G C
SspI  (9595)
1 site
A A T A T T T T A T A A
PvuI  (9161)
1 site
C G A T C G G C T A G C
AhdI  (8791)
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.
BmtI  (7517)
1 site
G C T A G C C G A T C G
NheI  (7513)
1 site
G C T A G C C G A T C G
PfoI  (6275)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
MluI  (5989)
1 site
A C G C G T T G C G C A
AfeI  (5928)
1 site
A G C G C T T C G C G A
RsrII  (5655)
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.
PflFI  (5257)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (5257)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
MscI  (5221)
1 site
T G G C C A A C C G G T
KflI  (1934)
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  (1934)
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.
BsrGI  (2229)
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.
AgeI  (2465)
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.
BsmBI  (3595)
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.
NdeI  (4249)
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.
TspMI  (4408)
1 site
C C C G G G G G G C C C
XmaI  (4408)
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.
SmaI  (4410)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (4417)
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.
AvrII  (4576)
1 site
C C T A G G G G A T C C
AarI  (4764)
1 site
C A C C T G C ( N ) 4 G T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
BmgBI  (4967)
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.
EF-1α promoter
2388 .. 3569  =  1182 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
2388 .. 3569  =  1182 bp
strong constitutive promoter for human elongation
factor EF-1α
AmpR
8718 .. 9578  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   8718 .. 9509  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8718 .. 9578  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   9510 .. 9578  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
8718 .. 9578  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
NeoR/KanR
5011 .. 5805  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
NeoR/KanR
5011 .. 5805  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from
Tn5
confers resistance to neomycin, kanamycin, and
G418 (Geneticin)
Tet-On 3G
3670 .. 4416  =  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
3670 .. 4416  =  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
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
3' LTR
6798 .. 7431  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
6798 .. 7431  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
6002 .. 6590  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
6002 .. 6590  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
7962 .. 8547  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7962 .. 8547  =  586 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
4425 .. 4997  =  573 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
4425 .. 4997  =  573 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
SV40 poly(A) signal
9731 .. 9865  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
9731 .. 9865  =  135 bp
SV40 polyadenylation signal
HIV-1 ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)
AmpR promoter
9579 .. 9683  =  105 bp
AmpR promoter
9579 .. 9683  =  105 bp
EF-1α intron A
2618 .. 3560  =  943 bp
intron upstream of the start codon
EF-1α intron A
2618 .. 3560  =  943 bp
intron upstream of the start codon
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