pLenti-C-Myc-DDK-IRES-GFP

Third-generation lentiviral PrecisionShuttle™ vector with a TurboGFP marker, for fusing an ORF to C-terminal c-Myc and FLAG® tags.

Sequence Author: OriGene

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BsmBI (7144) PasI (7137) ScaI (6955) BssSI (6374) DrdI (6309) Bsu36I (5625) BsrGI (4785) BsaBI * (4540) BlpI (4251) BstXI (4076) BmgBI (4033) SgrDI (1) CMV enhancer EcoNI (1550) BbvCI (1804) AleI (1957) XbaI (2613) CMV enhancer EcoRI (3279) BamHI (3292) AsiSI - SgfI (3324) MreI - SgrAI (3326) AscI (3330) NheI (3355) BmtI (3359) RsrII (3361) MluI (3367) BsiWI (3370) PaeR7I - PspXI - XhoI (3382) PmeI (3460) pLenti-C-Myc-DDK-IRES-GFP 7719 bp
BsmBI  (7144)
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.
BsmBI-v2 is an improved version of BsmBI.
PasI  (7137)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
ScaI  (6955)
1 site
A G T A C T T C A T G A
BssSI  (6374)
1 site
C A C G A G G T G C T C
DrdI  (6309)
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  (5625)
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  (4785)
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.
BsaBI  (4540)
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.
BlpI  (4251)
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.
BstXI  (4076)
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.
BmgBI  (4033)
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.
SgrDI  (1)
1 site
C G T C G A C G G C A G C T G C
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
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  (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
NheI  (3355)
1 site
G C T A G C C G A T C G
BmtI  (3359)
1 site
G C T A G C C G A T C 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.
PaeR7I  (3382)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3382)
1 site
V C T C G A G B B G A G C T C V
XhoI  (3382)
1 site
C T C G A G G A G C T C
PmeI  (3460)
1 site
G T T T A A A C C A A A T T T G
TurboGFP
4077 .. 4775  =  699 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
TurboGFP
4077 .. 4775  =  699 bp
232 amino acids  =  25.7 kDa
Product: improved green fluorescent protein from Pontellina plumata (Evdokimov et al., 2006)
mammalian codon-optimized
CmR
6927 .. 7586  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
6927 .. 7586  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
4903 .. 5491  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4903 .. 5491  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6262 .. 6850  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6262 .. 6850  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES2
3490 .. 4076  =  587 bp
3 segments
   Segment 1:  
   3490 .. 4064  =  575 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
3490 .. 4076  =  587 bp
3 segments
   Segment 2:  ATG  
   4065 .. 4067  =  3 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
3490 .. 4076  =  587 bp
3 segments
   Segment 3:  
   4068 .. 4076  =  9 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES2
3490 .. 4076  =  587 bp
3 segments
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
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)
6020 .. 6200  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
6020 .. 6200  =  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
7587 .. 7691  =  105 bp
AmpR promoter
7587 .. 7691  =  105 bp
loxP
4814 .. 4847  =  34 bp
Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT).
loxP
4814 .. 4847  =  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
Myc
3385 .. 3414  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
Myc
3385 .. 3414  =  30 bp
10 amino acids  =  1.2 kDa
Product: Myc (human c-Myc oncogene) epitope tag
FLAG
3433 .. 3456  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
FLAG
3433 .. 3456  =  24 bp
8 amino acids  =  1.0 kDa
Product: FLAG® epitope tag, followed by an enterokinase cleavage site
ORF:  1561 .. 2448  =  888 bp
ORF:  295 amino acids  =  33.8 kDa
ORF:  4549 .. 4827  =  279 bp
ORF:  92 amino acids  =  9.6 kDa
ORF:  4993 .. 5547  =  555 bp
ORF:  184 amino acids  =  20.5 kDa
ORF:  5006 .. 6046  =  1041 bp
ORF:  346 amino acids  =  38.3 kDa
ORF:  4065 .. 4775  =  711 bp
ORF:  236 amino acids  =  26.1 kDa
ORF:  1395 .. 1856  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  6927 .. 7586  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
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