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pLenti-C-Myc-DDK-IRES-tRFP

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

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pLenti-C-Myc-DDK-IRES-tRFP Sequence and MappLenti-C-Myc-DDK-IRES-tRFP.dna
Map and Sequence File   
Sequence Author:  OriGene
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 BsmBI (7120) DrdI (6285) SacII (5382) PshAI (4692) SfiI (4528) BmgBI (4033) AarI (3829) SgrDI (1) CMV enhancer KasI (1017) NarI (1018) SfoI (1019) PluTI (1021) BspQI - SapI (1482) FseI (1530) EcoNI (1550) BbvCI (1804) AleI (1957) PstI (2417) 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) PmlI (3806) pLenti-C-Myc-DDK-IRES-tRFP 7695 bp
BsmBI  (7120)
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.
DrdI  (6285)
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.
SacII  (5382)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
PshAI  (4692)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SfiI  (4528)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI
recognition sequence.
Sticky ends from different SfiI 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.
AarI  (3829)
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.
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.
FseI  (1530)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°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
PstI  (2417)
1 site
C T G C A G G A C G T 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
PmlI  (3806)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
TurboRFP
4065 .. 4760  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
TurboRFP
4065 .. 4760  =  696 bp
231 amino acids  =  26.1 kDa
Product: red fluorescent protein from Entacmaea
quadricolor

mammalian codon-optimized
CmR
6903 .. 7562  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
CmR
6903 .. 7562  =  660 bp
219 amino acids  =  25.7 kDa
Product: chloramphenicol acetyltransferase
confers resistance to chloramphenicol
WPRE
4879 .. 5467  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
WPRE
4879 .. 5467  =  589 bp
woodchuck hepatitis virus posttranscriptional
regulatory element
ori
6238 .. 6826  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
6238 .. 6826  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES
3513 .. 4064  =  552 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES
3513 .. 4064  =  552 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
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)
5996 .. 6176  =  181 bp
truncated 3' long terminal repeat (LTR) from HIV-1
3' LTR (truncated)
5996 .. 6176  =  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
7563 .. 7667  =  105 bp
AmpR promoter
7563 .. 7667  =  105 bp
loxP
4790 .. 4823  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
loxP
4790 .. 4823  =  34 bp
Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT).
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
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
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