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Plasmid Files

pLVX-EF1α-IRES-Puro

Lentiviral vector for bicistronic expression of a gene together with a puromycin-resistance marker.

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pLVX-EF1alpha-IRES-Puro Sequence and MappLVX-EF1alpha-IRES-Puro.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SgrDI (8505) SspI (8388) PvuI (7954) FspI (7806) AhdI (7584) DrdI (6799) BmtI (6308) NheI (6304) Bsu36I (5514) NcoI (5202) MluI (4781) SexAI * (4748) MscI * (4556) BstEII (4315) NruI * (833) KflI - PpuMI (1934) BspDI - ClaI (2180) SphI (2331) AgeI (2415) BstXI (2648) EcoRI (3535) SpeI (3547) XbaI (3553) NotI (3560) BamHI (3567) AvrII (3726) AarI (3914) BmgBI (4118) PflFI - Tth111I (4223) TspMI - XmaI (4230) SmaI (4232) BsiWI (4237) RsrII (4297) pLVX-EF1 α- IRES-Puro 8825 bp
SgrDI  (8505)
1 site
C G T C G A C G G C A G C T G C
SspI  (8388)
1 site
A A T A T T T T A T A A
PvuI  (7954)
1 site
C G A T C G G C T A G C
FspI  (7806)
1 site
T G C G C A A C G C G T
AhdI  (7584)
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.
DrdI  (6799)
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.
BmtI  (6308)
1 site
G C T A G C C G A T C G
NheI  (6304)
1 site
G C T A G C C G A T C G
Bsu36I  (5514)
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.
NcoI  (5202)
1 site
C C A T G G G G T A C C
MluI  (4781)
1 site
A C G C G T T G C G C A
SexAI  (4748)
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.
MscI  (4556)
1 site
T G G C C A A C C G G T
* Blocked by Dcm methylation.
BstEII  (4315)
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.
NruI  (833)
1 site
T C G C G A A G C G C T
* Blocked by Dam methylation.
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.
BspDI  (2180)
1 site
A T C G A T T A G C T A
ClaI  (2180)
1 site
A T C G A T T A G C T A
SphI  (2331)
1 site
G C A T G C C G T A C G
AgeI  (2415)
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  (2648)
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.
EcoRI  (3535)
1 site
G A A T T C C T T A A G
SpeI  (3547)
1 site
A C T A G T T G A T C A
XbaI  (3553)
1 site
T C T A G A A G A T C T
NotI  (3560)
1 site
G C G G C C G C C G C C G G C G
BamHI  (3567)
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  (3726)
1 site
C C T A G G G G A T C C
AarI  (3914)
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  (4118)
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.
PflFI  (4223)
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  (4223)
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.
TspMI  (4230)
1 site
C C C G G G G G G C C C
XmaI  (4230)
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  (4232)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BsiWI  (4237)
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.
RsrII  (4297)
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.
EF-1α promoter
2338 .. 3519  =  1182 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
2338 .. 3519  =  1182 bp
strong constitutive promoter for human elongation
factor EF-1α
AmpR
7511 .. 8371  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   7511 .. 8302  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7511 .. 8371  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   8303 .. 8371  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
7511 .. 8371  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
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
5589 .. 6222  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5589 .. 6222  =  634 bp
3' long terminal repeat (LTR) from HIV-1
PuroR
4181 .. 4780  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4181 .. 4780  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
4794 .. 5382  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4794 .. 5382  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6752 .. 7340  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6752 .. 7340  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
3575 .. 4148  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
3575 .. 4148  =  574 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.
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
8372 .. 8476  =  105 bp
AmpR promoter
8372 .. 8476  =  105 bp
MCS
3535 .. 3572  =  38 bp
multiple cloning site
MCS
3535 .. 3572  =  38 bp
multiple cloning site
EF-1α intron A
2568 .. 3510  =  943 bp
intron upstream of the start codon
EF-1α intron A
2568 .. 3510  =  943 bp
intron upstream of the start codon
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