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pLenti-EF1a-C-Myc-DDK-IRES-Puro

Lentiviral PrecisionShuttle™ vector with a puromycin resistance marker, for expression driven by the EF-1α promoter of an ORF fused to C-terminal c-Myc and FLAG® tags.

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pLenti-EF1a-C-Myc-DDK-IRES-Puro Sequence and MappLenti-EF1a-C-Myc-DDK-IRES-Puro.dna
Map and Sequence File   
Sequence Author:  OriGene
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 ScaI (7375) DrdI (6729) Bsu36I (6045) BsrGI (5205) SexAI * (5172) BstEII (4739) PflFI - Tth111I (4647) AarI (4369) PmlI (4346) SgrDI (1) SpeI (251) AseI (259) CMV enhancer NdeI (486) SnaBI (592) BspQI - SapI (1482) AleI (1957) XbaI (2614) AgeI (2703) EcoRI (3819) BamHI (3832) AsiSI - SgfI (3864) MreI - SgrAI (3866) AscI (3870) NheI (3895) BmtI (3899) MluI (3907) PspXI (3922) PmeI (4000) pLenti-EF1a-C-Myc-DDK-IRES-Puro 8139 bp
ScaI  (7375)
1 site
A G T A C T T C A T G A
DrdI  (6729)
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.
Bsu36I  (6045)
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.
BsrGI  (5205)
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.
SexAI  (5172)
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.
BstEII  (4739)
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.
PflFI  (4647)
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  (4647)
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.
AarI  (4369)
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.
PmlI  (4346)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
SgrDI  (1)
1 site
C G T C G A C G G C A G C T G C
SpeI  (251)
1 site
A C T A G T T G A T C A
AseI  (259)
1 site
A T T A A T T A A T T A
NdeI  (486)
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.
SnaBI  (592)
1 site
T A C G T A A T G C A T
BspQI  (1482)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1482)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
AleI  (1957)
1 site
C A C N N N N G T G G T G N N N N C A C
XbaI  (2614)
1 site
T C T A G A A G A T C T
AgeI  (2703)
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.
EcoRI  (3819)
1 site
G A A T T C C T T A A G
BamHI  (3832)
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.
AsiSI  (3864)
1 site
G C G A T C G C C G C T A G C G
SgfI  (3864)
1 site
G C G A T C G C C G C T A G C G
MreI  (3866)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (3866)
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.
AscI  (3870)
1 site
G G C G C G C C C C G C G C G G
NheI  (3895)
1 site
G C T A G C C G A T C G
BmtI  (3899)
1 site
G C T A G C C G A T C G
MluI  (3907)
1 site
A C G C G T T G C G C A
PspXI  (3922)
1 site
V C T C G A G B B G A G C T C V
PmeI  (4000)
1 site
G T T T A A A C C A A A T T T G
EF-1α promoter
2625 .. 3803  =  1179 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
2625 .. 3803  =  1179 bp
strong constitutive promoter for human elongation
factor EF-1α
CmR
7347 .. 8006  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
7347 .. 8006  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
PuroR
4605 .. 5204  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4605 .. 5204  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
WPRE
5323 .. 5911  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
5323 .. 5911  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6682 .. 7270  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6682 .. 7270  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
4030 .. 4603  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
4030 .. 4603  =  574 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
237 .. 616  =  380 bp
human cytomegalovirus immediate early enhancer
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
RRE
1683 .. 1916  =  234 bp
The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to
the cytoplasm.
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
618 .. 816  =  199 bp
human cytomegalovirus (CMV) immediate early
promoter
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
834 .. 1014  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
6440 .. 6620  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
6440 .. 6620  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
HIV-1 Ψ
1061 .. 1186  =  126 bp
packaging signal of human immunodeficiency virus
type 1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
cPPT/CTS
2443 .. 2560  =  118 bp
central polypurine tract and central termination
sequence of HIV-1
MCS
3819 .. 3928  =  110 bp
multiple cloning site
MCS
3819 .. 3928  =  110 bp
multiple cloning site
AmpR promoter
8007 .. 8111  =  105 bp
AmpR promoter
8007 .. 8111  =  105 bp
loxP
5234 .. 5267  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
5234 .. 5267  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
FLAG
3973 .. 3996  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
FLAG
3973 .. 3996  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an
enterokinase cleavage site
EF-1α intron A
2856 .. 3794  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
2856 .. 3794  =  939 bp
intron upstream of the start codon of human EF-1α
Myc
3925 .. 3954  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
3925 .. 3954  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
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