pRetroX-PTuner

Retroviral vector for fusing a destabilization domain to the N-terminus of a partner protein.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsViral Expression & Packaging VectorspRetroX-PTuner
NdeI (6006) SspI (5639) ScaI (5315) BspQI - SapI (3826) AgeI - SgrAI (2933) AscI (446) SpeI (729) PshAI (851) PstI (1014) MluI (1068) BtgZI (1138) EF-1α intron A BsgI (1395) NotI (1695) BglII (1702) BspDI - ClaI (1709) BamHI (1714) NcoI (1722) AvrII (1881) PflMI (2183) BmgBI (2273) BsiWI (2392) SalI (2444) AccI (2445) HincII (2446) RsrII (2452) BstEII (2470) StuI (2667) CsiI - SexAI * (2903) pRetroX-PTuner 6229 bp
NdeI  (6006)
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  (5639)
1 site		 A A T A T T T T A T A A
ScaI  (5315)
1 site		 A G T A C T T C A T G A
BspQI  (3826)
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  (3826)
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.
AgeI  (2933)
1 site		 A C C G G T T G G C C A
SgrAI  (2933)
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  (446)
1 site		 G G C G C G C C C C G C G C G G
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
BtgZI  (1138)
1 site		 G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4
Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
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.
NcoI  (1722)
1 site		 C C A T G G G G T A C C
AvrII  (1881)
1 site		 C C T A G G G G A T C C
PflMI  (2183)
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.
BmgBI  (2273)
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.
BsiWI  (2392)
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.
SalI  (2444)
1 site		 G T C G A C C A G C T G
AccI  (2445)
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  (2446)
1 site		 G T Y R A C C A R Y T G
RsrII  (2452)
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.
BstEII  (2470)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
StuI  (2667)
1 site		 A G G C C T T C C G G A
CsiI  (2903)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI sites may not be compatible.
SexAI  (2903)
1 site		 A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AmpR
4762 .. 5622  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4762 .. 5553  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4762 .. 5622  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   5554 .. 5622  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4762 .. 5622  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
PuroR
2336 .. 2932  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
2336 .. 2932  =  597 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
LTR
2992 .. 3585  =  594 bp
long terminal repeat from Moloney murine leukemia virus
LTR
2992 .. 3585  =  594 bp
long terminal repeat from Moloney murine leukemia virus
LTR
1 .. 592  =  592 bp
long terminal repeat from Moloney murine leukemia virus
LTR
1 .. 592  =  592 bp
long terminal repeat from Moloney murine leukemia virus
ori
4003 .. 4591  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4003 .. 4591  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
1730 .. 2303  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
1730 .. 2303  =  574 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
5623 .. 5727  =  105 bp
AmpR promoter
5623 .. 5727  =  105 bp
MCS
1694 .. 1727  =  34 bp
multiple cloning site
MCS
1694 .. 1727  =  34 bp
multiple cloning site
ORF:  2213 .. 2959  =  747 bp
ORF:  248 amino acids  =  27.0 kDa
ORF:  4892 .. 5158  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  6008 .. 3  =  225 bp
ORF:  74 amino acids  =  8.7 kDa
ORF:  1368 .. 1781  =  414 bp
ORF:  137 amino acids  =  15.4 kDa
ORF:  734 .. 1018  =  285 bp
ORF:  94 amino acids  =  9.8 kDa
ORF:  4762 .. 5622  =  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
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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