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.

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.
BsmBI-v2 is an improved version of BsmBI.
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

Cleavage may be enhanced when more than one copy of the AarI recognition sequence is present.
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
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).
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:  4969 .. 5523  =  555 bp
ORF:  184 amino acids  =  20.5 kDa
ORF:  4982 .. 6022  =  1041 bp
ORF:  346 amino acids  =  38.3 kDa
ORF:  4065 .. 4760  =  696 bp
ORF:  231 amino acids  =  26.1 kDa
ORF:  1395 .. 1856  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  6903 .. 7562  =  660 bp
ORF:  219 amino acids  =  25.7 kDa
ORF:  4166 .. 4411  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
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