pRetroX-PTuner IRES

Retroviral vector for fusing a destabilization domain to the N-terminus of a partner protein, with co-expression of ZsGreen1.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsViral Expression & Packaging VectorspRetroX-PTuner IRES
NdeI (6060) SspI (5693) ScaI (5369) BspQI - SapI (3880) AscI (446) MscI (661) SpeI (729) PshAI (851) PstI (1014) MluI (1068) EF-1α intron A BsgI (1395) NotI (1695) BglII (1702) BspDI - ClaI (1709) BamHI (1714) AvrII (1874) PflMI (2176) BclI * (2379) SgrAI (2384) SalI (2534) AccI (2535) HincII (2536) NgoMIV (2561) NaeI (2563) BsrGI (2847) pRetroX-PTuner IRES 6283 bp
NdeI  (6060)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SspI  (5693)
1 site		 A A T A T T T T A T A A
ScaI  (5369)
1 site		 A G T A C T T C A T G A
BspQI  (3880)
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  (3880)
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.
AscI  (446)
1 site		 G G C G C G C C C C G C G C G G
MscI  (661)
1 site		 T G G C C A A C C G G T
SpeI  (729)
1 site		 A C T A G T T G A T C A
PshAI  (851)
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.
PstI  (1014)
1 site		 C T G C A G G A C G T C
MluI  (1068)
1 site		 A C G C G T T G C G C A
BsgI  (1395)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
NotI  (1695)
1 site		 G C G G C C G C C G C C G G C G
BglII  (1702)
1 site		 A G A T C T T C T A G A
BspDI  (1709)
1 site		 A T C G A T T A G C T A
ClaI  (1709)
1 site		 A T C G A T T A G C T A
BamHI  (1714)
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  (1874)
1 site		 C C T A G G G G A T C C
PflMI  (2176)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
BclI  (2379)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
SgrAI  (2384)
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.
SalI  (2534)
1 site		 G T C G A C C A G C T G
AccI  (2535)
1 site		 G T M K A C C A K M T G
Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HincII  (2536)
1 site		 G T Y R A C C A R Y T G
NgoMIV  (2561)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (2563)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
BsrGI  (2847)
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.
AmpR
4816 .. 5676  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4816 .. 5607  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4816 .. 5676  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5608 .. 5676  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4816 .. 5676  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ZsGreen1
2297 .. 2992  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
2297 .. 2992  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
LTR
3046 .. 3639  =  594 bp
long terminal repeat from Moloney murine leukemia virus
LTR
3046 .. 3639  =  594 bp
long terminal repeat from Moloney murine leukemia virus
ori
4057 .. 4645  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4057 .. 4645  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
1723 .. 2295  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
1723 .. 2295  =  573 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
MMLV Ψ
655 .. 1012  =  358 bp
packaging signal of Moloney murine leukemia virus (MMLV)
MMLV Ψ
655 .. 1012  =  358 bp
packaging signal of Moloney murine leukemia virus (MMLV)
DD
1368 .. 1691  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 1:  
   1368 .. 1370  =  3 bp
   1 amino acid  =  149.2 Da
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
1368 .. 1691  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
   Segment 2:  
   1371 .. 1691  =  321 bp
   107 amino acids  =  11.8 kDa
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
1368 .. 1691  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
Product: destabilization domain that can be stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
EF-1α intron A
1075 .. 1278  =  204 bp
truncated intron upstream of the start codon of human EF-1α
EF-1α intron A
1075 .. 1278  =  204 bp
truncated intron upstream of the start codon of human EF-1α
EF-1α exon
1279 .. 1308  =  30 bp
truncated exon 2 from human EF-1α, beginning with a GT-to-CC mutation that reduces splicing efficiency
EF-1α exon
1279 .. 1308  =  30 bp
truncated exon 2 from human EF-1α, beginning with a GT-to-CC mutation that reduces splicing efficiency
AmpR promoter
5677 .. 5781  =  105 bp
AmpR promoter
5677 .. 5781  =  105 bp
MCS
1694 .. 1719  =  26 bp
multiple cloning site
MCS
1694 .. 1719  =  26 bp
multiple cloning site
ORF:  2297 .. 2992  =  696 bp
ORF:  231 amino acids  =  26.1 kDa
ORF:  4946 .. 5212  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6062 .. 3  =  225 bp
ORF:  74 amino acids  =  8.7 kDa
ORF:  1368 .. 1835  =  468 bp
ORF:  155 amino acids  =  17.3 kDa
ORF:  734 .. 1018  =  285 bp
ORF:  94 amino acids  =  9.8 kDa
ORF:  2674 .. 2967  =  294 bp
ORF:  97 amino acids  =  10.9 kDa
ORF:  4816 .. 5676  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
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Individual Sequences & Maps

pAAV-AC-GFP
pLenti6.2 N-Lumio V5-DEST
pLVX-rHom-1
pRetroX-Tet3G
pAAV-AC-Myc-DDK
pLenti6.2 V5-DEST
pLVX-rHom-Nuc1
pRetroX-TetOne
pAAV-AC-RFP
pLenti6.3 TO V5-DEST
pLVX-rHom-Sec1
pRetroX-TetOne-Puro
pAAV2-EF1a-tGFP-WPRE
pLenti6.3 V5-DEST
pLVX-Tet-Off Advanced
pRetroX-Tight-Hyg
pAd BLOCK-iT-DEST
pLenti6.4 R4R2 V5-DEST
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
pRSI16-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pAdenoX-ZsGreen1 Linear
pLHCX
pLVX-TRE3G-ZsGreen1
pRSI16cb-U6-sh-13kCB18-HTS6-UbiC-TagRFP-2A-Puro
pB-RS
pLKO.1
pLX301
pRSI17-U6-sh-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Hygro
pLKO.1 puro
pLX302
pRSI17cb-U6-sh-13kCB18-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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