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pLenti-EF1a-C-tGFP

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

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pLenti-EF1a-C-tGFP.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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BsmBI (7007) MscI (6968) ScaI (6818) DrdI (6172) Bsu36I (5488) PfoI (5039) BsrGI (4648) PmeI (4629) BsaBI * (4390) AarI (4378) BlpI (4101) NotI (3915) BsiWI (3909) MluI (3906) RsrII (3900) AscI (3869) MreI - SgrAI (3865) AsiSI - SgfI (3863) BamHI (3831) SgrDI (1) AseI (259) NdeI (486) SnaBI (592) AleI (1957) SanDI (2314) XbaI (2613) BmgBI (2619) AgeI (2702) EcoRI (3818) pLenti-EF1a-C-tGFP 7582 bp
BsmBI  (7007)
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  (6968)
1 site
T G G C C A A C C G G T
ScaI  (6818)
1 site
A G T A C T T C A T G A
DrdI  (6172)
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  (5488)
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.
PfoI  (5039)
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.
BsrGI  (4648)
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.
PmeI  (4629)
1 site
G T T T A A A C C A A A T T T G
BsaBI  (4390)
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.
AarI  (4378)
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.
BlpI  (4101)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
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
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.
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
AleI  (1957)
1 site
C A C N N N N G T G G T G N N N N C A C
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
BmgBI  (2619)
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.
AgeI  (2702)
1 site
A C C G G T T G G C C A
EcoRI  (3818)
1 site
G A A T T C C T T A A G
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
TurboGFP
3927 .. 4622  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
3927 .. 4622  =  696 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
CmR
6790 .. 7449  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
6790 .. 7449  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
4766 .. 5354  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4766 .. 5354  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6125 .. 6713  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6125 .. 6713  =  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)
5883 .. 6063  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
5883 .. 6063  =  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
7450 .. 7554  =  105 bp
AmpR promoter
7450 .. 7554  =  105 bp
MCS
3818 .. 3921  =  104 bp
multiple cloning site
MCS
3818 .. 3921  =  104 bp
multiple cloning site
loxP
4677 .. 4710  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
4677 .. 4710  =  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
ORF:  1561 .. 2448  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  3259 .. 3492  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  4399 .. 4677  =  279 bp
ORF:  92 amino acids  =  9.7 kDa
ORF:  4856 .. 5410  =  555 bp
ORF:  184 amino acids  =  20.5 kDa
ORF:  3180 .. 3617  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  3927 .. 4631  =  705 bp
ORF:  234 amino acids  =  25.9 kDa
ORF:  4869 .. 5909  =  1041 bp
ORF:  346 amino acids  =  38.3 kDa
ORF:  6790 .. 7449  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  1395 .. 1856  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  2922 .. 3542  =  621 bp
ORF:  206 amino acids  =  23.0 kDa
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