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Plasmid Files

pLenti-C-mGFP

Third-generation lentiviral vector for expression and monomeric GFP tagging of an ORF. Can be used as an entry vector for the PrecisionShuttle™ system.

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pLenti-C-mGFP.dna
Map and Sequence File:    Download    Open   
Sequence Author:  OriGene
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AclI (6637) BsmBI (6498) MscI (6459) ScaI (6309) PspFI (5863) BseYI (5859) DrdI (5663) Bsu36I (4979) PmeI (4120) BsaBI * (3973) BstEII (3567) SgrDI (1) KasI (1017) NarI (1018) SfoI (1019) PluTI (1021) BspQI - SapI (1482) EcoNI (1550) BbvCI (1804) SanDI (2314) PstI (2417) PspOMI (2605) ApaI (2609) XbaI (2613) EcoRI (3279) BamHI (3292) AsiSI - SgfI (3324) MreI - SgrAI (3326) AscI (3330) RsrII (3361) MluI (3367) BsiWI (3370) NotI (3376) PaeR7I - XhoI (3382) BmgBI (3450) pLenti-C-mGFP 7073 bp
AclI  (6637)
1 site
A A C G T T T T G C A A
BsmBI  (6498)
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  (6459)
1 site
T G G C C A A C C G G T
ScaI  (6309)
1 site
A G T A C T T C A T G A
PspFI  (5863)
1 site
C C C A G C G G G T C G
BseYI  (5859)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
DrdI  (5663)
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  (4979)
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  (4120)
1 site
G T T T A A A C C A A A T T T G
BsaBI  (3973)
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  (3567)
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.
SgrDI  (1)
1 site
C G T C G A C G G C A G C T G C
KasI  (1017)
1 site
G G C G C C C C G C G G
NarI  (1018)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (1019)
1 site
G G C G C C C C G C G G
PluTI  (1021)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
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.
EcoNI  (1550)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BbvCI  (1804)
1 site
C C T C A G C G G A G T C G
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.
PstI  (2417)
1 site
C T G C A G G A C G T C
PspOMI  (2605)
1 site
G G G C C C C C C G G G
ApaI  (2609)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
XbaI  (2613)
1 site
T C T A G A A G A T C T
EcoRI  (3279)
1 site
G A A T T C C T T A A G
BamHI  (3292)
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  (3324)
1 site
G C G A T C G C C G C T A G C G
SgfI  (3324)
1 site
G C G A T C G C C G C T A G C G
MreI  (3326)
1 site
C G C C G G C G G C G G C C G C
SgrAI  (3326)
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  (3330)
1 site
G G C G C G C C C C G C G C G G
RsrII  (3361)
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.
MluI  (3367)
1 site
A C G C G T T G C G C A
BsiWI  (3370)
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.
NotI  (3376)
1 site
G C G G C C G C C G C C G G C G
PaeR7I  (3382)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (3382)
1 site
C T C G A G G A G C T C
BmgBI  (3450)
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.
TagGFP2
3388 .. 4101  =  714 bp
238 amino acids  =  26.9 kDa
Product: monomeric green fluorescent protein
mammalian codon-optimized
TagGFP2
3388 .. 4101  =  714 bp
238 amino acids  =  26.9 kDa
Product: monomeric green fluorescent protein
mammalian codon-optimized
CmR
6281 .. 6940  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
6281 .. 6940  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
4257 .. 4845  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4257 .. 4845  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
5616 .. 6204  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5616 .. 6204  =  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
CMV enhancer
2643 .. 3022  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2643 .. 3022  =  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
3023 .. 3226  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
3023 .. 3226  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
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)
5374 .. 5554  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
5374 .. 5554  =  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
3279 .. 3387  =  109 bp
multiple cloning site
MCS
3279 .. 3387  =  109 bp
multiple cloning site
AmpR promoter
6941 .. 7045  =  105 bp
AmpR promoter
6941 .. 7045  =  105 bp
loxP
4168 .. 4201  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
4168 .. 4201  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
T7 promoter
3252 .. 3270  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
3252 .. 3270  =  19 bp
promoter for bacteriophage T7 RNA polymerase
ORF:  1561 .. 2448  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  3388 .. 4116  =  729 bp
ORF:  242 amino acids  =  27.3 kDa
ORF:  4360 .. 5400  =  1041 bp
ORF:  346 amino acids  =  38.3 kDa
ORF:  4347 .. 4901  =  555 bp
ORF:  184 amino acids  =  20.5 kDa
ORF:  1395 .. 1856  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  3660 .. 3899  =  240 bp
ORF:  79 amino acids  =  9.4 kDa
ORF:  6281 .. 6940  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
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