pLVX-EF1alpha-IRES-mCherry

Lentiviral bicistronic expression vector for constitutively co-expressing a protein of interest together with mCherry from the EF-1α promoter.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsViral Expression & Packaging VectorspLVX-EF1alpha-IRES-mCherry
SgrDI (8584) SspI (8467) PvuI (8033) FspI (7885) BmtI (6387) NheI (6383) Bsu36I (5593) PfoI (5146) MluI (4860) BsrGI (4849) XcmI (4835) SgrAI (4830) SbfI (4504) BssHII (711) NruI * (833) KflI - PpuMI (1934) BspDI - ClaI (2180) SphI (2331) AgeI (2415) BstXI (2648) EcoRI (3535) SpeI (3547) XbaI (3553) NotI (3560) BamHI (3567) AvrII (3726) PaqCI (3914) BmgBI (4117) MscI (4179) pLVX-EF1 α- IRES-mCherry 8904 bp
SgrDI  (8584)
1 site		 C G T C G A C G G C A G C T G C
SspI  (8467)
1 site		 A A T A T T T T A T A A
PvuI  (8033)
1 site		 C G A T C G G C T A G C
FspI  (7885)
1 site		 T G C G C A A C G C G T
BmtI  (6387)
1 site		 G C T A G C C G A T C G
NheI  (6383)
1 site		 G C T A G C C G A T C G
Bsu36I  (5593)
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  (5146)
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.
MluI  (4860)
1 site		 A C G C G T T G C G C A
BsrGI  (4849)
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.
XcmI  (4835)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
SgrAI  (4830)
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.
SbfI  (4504)
1 site		 C C T G C A G G G G A C G T C C
BssHII  (711)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
NruI  (833)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
KflI  (1934)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different KflI sites may not be compatible.
PpuMI  (1934)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
BspDI  (2180)
1 site		 A T C G A T T A G C T A
ClaI  (2180)
1 site		 A T C G A T T A G C T A
SphI  (2331)
1 site		 G C A T G C C G T A C G
AgeI  (2415)
1 site		 A C C G G T T G G C C A
BstXI  (2648)
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.
EcoRI  (3535)
1 site		 G A A T T C C T T A A G
SpeI  (3547)
1 site		 A C T A G T T G A T C A
XbaI  (3553)
1 site		 T C T A G A A G A T C T
NotI  (3560)
1 site		 G C G G C C G C C G C C G G C G
BamHI  (3567)
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.
AvrII  (3726)
1 site		 C C T A G G G G A T C C
PaqCI  (3914)
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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
BmgBI  (4117)
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.
MscI  (4179)
1 site		 T G G C C A A C C G G T
EF-1α promoter
2338 .. 3519  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
EF-1α promoter
2338 .. 3519  =  1182 bp
strong constitutive promoter for human elongation factor EF-1α
AmpR
7590 .. 8450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7590 .. 8381  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7590 .. 8450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   8382 .. 8450  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7590 .. 8450  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
mCherry
4149 .. 4859  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
mCherry
4149 .. 4859  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)
mammalian codon-optimized
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
1 .. 634  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5668 .. 6301  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5668 .. 6301  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
4873 .. 5461  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
4873 .. 5461  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
6831 .. 7419  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
6831 .. 7419  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
3575 .. 4147  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
3575 .. 4147  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1303 .. 1536  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
SV40 poly(A) signal
8603 .. 8737  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8603 .. 8737  =  135 bp
SV40 polyadenylation signal
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
681 .. 806  =  126 bp
packaging signal of human immunodeficiency virus type 1
cPPT/CTS
2028 .. 2143  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2143  =  116 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
8451 .. 8555  =  105 bp
AmpR promoter
8451 .. 8555  =  105 bp
MCS
3535 .. 3572  =  38 bp
multiple cloning site
MCS
3535 .. 3572  =  38 bp
multiple cloning site
EF-1α intron A
2568 .. 3510  =  943 bp
intron upstream of the start codon of human EF-1α
EF-1α intron A
2568 .. 3510  =  943 bp
intron upstream of the start codon of human EF-1α
ORF:  4963 .. 5490  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  7720 .. 7986  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  2897 .. 3334  =  438 bp
ORF:  145 amino acids  =  15.9 kDa
ORF:  4976 .. 5272  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  2976 .. 3209  =  234 bp
ORF:  77 amino acids  =  7.7 kDa
ORF:  4149 .. 4859  =  711 bp
ORF:  236 amino acids  =  26.7 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  7590 .. 8450  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2612 .. 3259  =  648 bp
ORF:  215 amino acids  =  23.7 kDa
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Individual Sequences & Maps

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pAAV2-EF1a-tGFP-WPRE
pLenti6.3 V5-DEST
pLVX-Tet-Off Advanced
pRetroX-Tight-Hyg
pAd BLOCK-iT-DEST
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pLVX-Tet-On Advanced
pRetroX-Tight-Pur
pAd CMV V5-DEST
pLenti6 BLOCK-iT-DEST
pLVX-Tet3G
pRetroX-TRE3G
pAd PL-DEST
pLenti6 capTEV-CT-DEST
pLVX-TetOne
pRFP-C-RS
pAdenoX-CMV Linear
pLenti6 capTEV-NT-DEST1
pLVX-TetOne-Puro
pRFP-CB-shLenti
pAdenoX-DsRed-Express Linear
pLenti6 TR
pLVX-Tight-Puro
pRS
pAdenoX-PRLS Linear
pLenti6 UbC V5-DEST
pLVX-TRE3G
pRSGC1-U6-sg-HTS6-CMV-Cas9-2A-Puro
pAdenoX-PRLS-DsRed-Express Linear
pLenti6 V5-DEST
pLVX-TRE3G-Hom1
pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro
pAdenoX-PRLS-ZsGreen1 Linear
pLenti7.3 V5-DEST
pLVX-TRE3G-IRES
pRSI12-U6-sh-HTS4-UbiC-TagRFP-2A-Puro
pAdenoX-Tet3G Linear
pLEX-MCS
pLVX-TRE3G-mCherry
pRSI16cb-U6-sh-13kCB18-HTS6-UbiC-TagRFP-2A-Puro
pAdenoX-ZsGreen1 Linear
pLHCX
pLVX-TRE3G-ZsGreen1
pRSI16-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pB-RS
pLKO.1
pLX301
pRSI17cb-U6-sh-13kCB18-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Hygro
pLKO.1 puro
pLX302
pRSI17-U6-sh-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Neo
pLMN-ZsGreen-Neomycin
pLX303
pRSIT12-U6Tet-sh-HTS4-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Puro
pLMPd-Ametrine
pLX304
pRSIT16-U6Tet-sh-HTS6-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Zeo
pLNCX2
pLX304-V5-Blast-Empty
pRSIT17-U6Tet-sh-HTS6-CMV-TetRep-2A-TagGFP2-2A-Puro
pCMV-VSV-G
pLP VSVG
pLXIN
pRSITEP-U6Tet-sh-noHTS-EF1-TetRep-2A-Puro
pGFP-B-RS
pLP1
pLXSN
pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP
pGFP-C-shLenti
pLP2
pMD2.G
pShuttle2
pGFP-V-RS
pLPCX
pMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pSIREN-RetroQ
pGIPZ
pLVX-EF1alpha-IRES-mCherry
pMSCVhyg
pSIREN-RetroQ-DsRed-Express
pLenti-C-GFP
pLVX-EF1alpha-IRES-Puro
pMSCVneo
pSIREN-RetroQ-TetH
pLenti-C-mGFP
pLVX-EF1alpha-IRES-ZsGreen1
pMSCVpuro
pSIREN-RetroQ-TetP
pLenti-C-Myc-DDK
pLVX-EF1alpha-Tet3G
pQC-tTS-IN
pSIREN-RetroQ-ZsGreen1
pLenti-C-Myc-DDK-IRES-BSD
pLVX-Het-1
pQCXIH
psPAX2
pLenti-C-Myc-DDK-IRES-GFP
pLVX-Het-2
pQCXIN
pTRE3G-Hom1
pLenti-C-Myc-DDK-IRES-Neo
pLVX-Het-Mem1
pQCXIP
pTRIPZ
pLenti-C-Myc-DDK-IRES-Puro
pLVX-Het-Nuc1
pQCXIX
ptTS-Neo
pLenti-C-Myc-DDK-IRES-tRFP
pLVX-Hom-1
pRetro-Lib
pZIP-hCMV-RFP-Puro
pLenti-C-tRFP
pLVX-Hom-Mem1
pRetroQ-AcGFP1-C1
pZIP-hCMV-ZSGreen-Puro
pLenti-C-tYFP
pLVX-Hom-Nuc1
pRetroQ-AcGFP1-N1
pZIP-hEF1-alpha-RFP-Puro
pLenti-EF1a-C-mGFP
pLVX-IRES-Hyg
pRetroQ-DsRed-Monomer-C1
pZIP-hEF1-alpha-ZsGreen-Puro
pLenti-EF1a-C-Myc-DDK-IRES-Puro
pLVX-IRES-mCherry
pRetroQ-DsRed-Monomer-N1
pZIP-mCMV-RFP-Puro
pLenti-EF1a-C-tGFP
pLVX-IRES-Neo
pRetroQ-mCherry-C1
pZIP-mCMV-ZsGreen-Puro
pLenti-N-tGFP
pLVX-IRES-Puro
pRetroQ-mCherry-N1
pZIP-mEF1-alpha-RFP-Puro
pLenti-N-tRFP
pLVX-IRES-tdTomato
pRetroX-IRES-DsRedExpress
pZIP-mEF1-alpha-ZsGreen-Puro
pLenti-N-tYFP
pLVX-IRES-ZsGreen1
pRetroX-IRES-ZsGreen1
pZIP-SFFV-RFP-Puro
pLenti3.3 TR
pLVX-PTuner
pRetroX-PTuner
pZIP-SFFV-ZsGreen-Puro
pLenti4 BLOCK-iT-DEST
pLVX-PTuner Green
pRetroX-PTuner IRES
pZIP-TRE3G-ZsGreen-Puro
pLenti4 V5-DEST
pLVX-PTunerC
pRetroX-Tet-Off Advanced
pSIREN-Shuttle (linearized)
pLenti6.2 C-Lumio V5-DEST
pLVX-Puro
pRetroX-Tet-On Advanced
scramble shRNA

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