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Plasmid Files

pLenti-EF1a-C-mGFP

Lentiviral PrecisionShuttle™ vector for expression driven by the EF-1α promoter of an ORF tagged at the C-terminus with TagGFP2 (mTagGFP).

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pLenti-EF1a-C-mGFP Sequence and MappLenti-EF1a-C-mGFP.dna
Map and Sequence File   
Sequence Author:  OriGene
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 BsmBI (7037) MscI (6998) ScaI (6848) DrdI (6202) Bsu36I (5518) PmeI (4659) BsaBI * (4512) BstEII (4106) NotI (3915) BsiWI (3909) MluI (3906) RsrII (3900) AscI (3869) MreI - SgrAI (3865) AsiSI - SgfI (3863) SgrDI (1) AseI (259) CMV enhancer NdeI (486) SnaBI (592) BspQI - SapI (1482) SanDI (2314) XbaI (2613) AgeI (2702) BfuAI - BspMI (3148) EcoRI (3818) BamHI (3831) pLenti-EF1a-C-mGFP 7612 bp
BsmBI  (7037)
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.
MscI  (6998)
1 site
T G G C C A A C C G G T
ScaI  (6848)
1 site
A G T A C T T C A T G A
DrdI  (6202)
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  (5518)
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.
PmeI  (4659)
1 site
G T T T A A A C C A A A T T T G
BsaBI  (4512)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
BstEII  (4106)
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.
NotI  (3915)
1 site
G C G G C C G C C G C C G G C G
BsiWI  (3909)
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.
MluI  (3906)
1 site
A C G C G T T G C G C A
RsrII  (3900)
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.
AscI  (3869)
1 site
G G C G C G C C C C G C G C G G
MreI  (3865)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (3865)
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.
AsiSI  (3863)
1 site
G C G A T C G C C G C T A G C G
SgfI  (3863)
1 site
G C G A T C G C C G C T A G C G
SgrDI  (1)
1 site
C G T C G A C G G C A G C T G C
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.
SanDI  (2314)
1 site
G G G W C C C C C C W G G G

Sticky ends from different SanDI sites may not be compatible.
XbaI  (2613)
1 site
T C T A G A A G A T C T
AgeI  (2702)
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.
BfuAI  (3148)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (3148)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
EcoRI  (3818)
1 site
G A A T T C C T T A A G
BamHI  (3831)
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.
EF-1α promoter
2624 .. 3802  =  1179 bp
strong constitutive promoter for human elongation
factor EF-1α
EF-1α promoter
2624 .. 3802  =  1179 bp
strong constitutive promoter for human elongation
factor EF-1α
TagGFP2
3927 .. 4640  =  714 bp
238 amino acids  =  26.9 kDa
Product: monomeric green fluorescent protein, also
known as mTagGFP
mammalian codon-optimized
TagGFP2
3927 .. 4640  =  714 bp
238 amino acids  =  26.9 kDa
Product: monomeric green fluorescent protein, also
known as mTagGFP
mammalian codon-optimized
CmR
6820 .. 7479  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
6820 .. 7479  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
4796 .. 5384  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4796 .. 5384  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6155 .. 6743  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6155 .. 6743  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
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)
5913 .. 6093  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
5913 .. 6093  =  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
AmpR promoter
7480 .. 7584  =  105 bp
AmpR promoter
7480 .. 7584  =  105 bp
MCS
3818 .. 3921  =  104 bp
multiple cloning site
MCS
3818 .. 3921  =  104 bp
multiple cloning site
loxP
4707 .. 4740  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
4707 .. 4740  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
EF-1α intron A
2855 .. 3793  =  939 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
2855 .. 3793  =  939 bp
intron upstream of the start codon of human EF-1α
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