pRetroX-Tight-Hyg

Retroviral vector for expressing a gene with the Tet-On® Advanced or Tet-Off® Advanced system.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsViral Expression & Packaging VectorspRetroX-Tight-Hyg
SspI (7074) XmnI (6869) FspI (6492) PciI (5377) BspQI - SapI (5261) AvrII (5130) SfiI (5083) PvuII (4056) EcoRV (4013) BspDI - ClaI (3828) AleI (3741) CMV enhancer SnaBI (284) AscI (542) AfeI (1088) BstEII (1252) BglII (1576) sequencing primer (1578 .. 1601) PaeR7I - PspXI - XhoI (1600) BamHI (1925) NotI (1932) NgoMIV (1939) NaeI (1941) XbaI * (1945) NruI * (1953) MluI (1957) EcoRI (1963) AgeI (2083) BsmI (2404) AsiSI (2854) RsrII (2898) BbvCI (3555) pRetroX-Tight-Hyg 7419 bp
SspI  (7074)
1 site		 A A T A T T T T A T A A
XmnI  (6869)
1 site		 G A A N N N N T T C C T T N N N N A A G
FspI  (6492)
1 site		 T G C G C A A C G C G T
PciI  (5377)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
BspQI  (5261)
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  (5261)
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.
AvrII  (5130)
1 site		 C C T A G G G G A T C C
SfiI  (5083)
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.
PvuII  (4056)
1 site		 C A G C T G G T C G A C
EcoRV  (4013)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
BspDI  (3828)
1 site		 A T C G A T T A G C T A
ClaI  (3828)
1 site		 A T C G A T T A G C T A
AleI  (3741)
1 site		 C A C N N N N G T G G T G N N N N C A C
SnaBI  (284)
1 site		 T A C G T A A T G C A T
AscI  (542)
1 site		 G G C G C G C C C C G C G C G G
AfeI  (1088)
1 site		 A G C G C T T C G C G A
BstEII  (1252)
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.
BglII  (1576)
1 site		 A G A T C T T C T A G A
PaeR7I  (1600)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1600)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1600)
1 site		 C T C G A G G A G C T C
BamHI  (1925)
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.
NotI  (1932)
1 site		 G C G G C C G C C G C C G G C G
NgoMIV  (1939)
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  (1941)
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.
XbaI  (1945)
1 site		 T C T A G A A G A T C T
* Blocked by Dam methylation.
NruI  (1953)
1 site		 T C G C G A A G C G C T
* Blocked by Dam methylation.
MluI  (1957)
1 site		 A C G C G T T G C G C A
EcoRI  (1963)
1 site		 G A A T T C C T T A A G
AgeI  (2083)
1 site		 A C C G G T T G G C C A
BsmI  (2404)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
AsiSI  (2854)
1 site		 G C G A T C G C C G C T A G C G
RsrII  (2898)
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.
BbvCI  (3555)
1 site		 C C T C A G C G G A G T C G
sequencing primer
24-mer  /  50% GC
1 binding site
1578 .. 1601  =  24 annealed bases
Tm  =  62°C
PTight sequencing primer
HygR
2490 .. 3515  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
HygR
2490 .. 3515  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
Amp
6197 .. 7057  =  861 bp
286 amino acids  =  31.5 kDa
Amp
6197 .. 7057  =  861 bp
286 amino acids  =  31.5 kDa
ori
5438 .. 6026  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5438 .. 6026  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
PGK promoter
1973 .. 2472  =  500 bp
mouse phosphoglycerate kinase 1 promoter
PGK promoter
1973 .. 2472  =  500 bp
mouse phosphoglycerate kinase 1 promoter
3' LTR (ΔU3)
4125 .. 4550  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus
3' LTR (ΔU3)
4125 .. 4550  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus
gag (truncated)
1151 .. 1567  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
1151 .. 1567  =  417 bp
truncated MMLV gag gene lacking the start codon
CMV enhancer
7348 .. 308  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
7348 .. 308  =  380 bp
human cytomegalovirus immediate early enhancer
SV40 promoter
4816 .. 5145  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4816 .. 5145  =  330 bp
SV40 enhancer and early promoter
tight TRE promoter
1606 .. 1917  =  312 bp
Tet-responsive promoter PTight
tight TRE promoter
1606 .. 1917  =  312 bp
Tet-responsive promoter PTight
CMV promoter
309 .. 512  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
309 .. 512  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
MMLV Ψ
751 .. 950  =  200 bp
packaging signal of Moloney murine leukemia virus (MMLV)
MMLV Ψ
751 .. 950  =  200 bp
packaging signal of Moloney murine leukemia virus (MMLV)
5' LTR (truncated)
513 .. 688  =  176 bp
truncated long terminal repeat from Moloney murine sarcoma virus
5' LTR (truncated)
513 .. 688  =  176 bp
truncated long terminal repeat from Moloney murine sarcoma virus
AmpR promoter
7058 .. 7162  =  105 bp
AmpR promoter
7058 .. 7162  =  105 bp
MCS
1925 .. 1968  =  44 bp
multiple cloning site
MCS
1925 .. 1968  =  44 bp
multiple cloning site
SV40 ori
4996 .. 5131  =  136 bp
SV40 origin of replication
SV40 ori
4996 .. 5131  =  136 bp
SV40 origin of replication
tet operator
1611 .. 1629  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1611 .. 1629  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1647 .. 1665  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1647 .. 1665  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1682 .. 1700  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1682 .. 1700  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1718 .. 1736  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1718 .. 1736  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1754 .. 1772  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1754 .. 1772  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1789 .. 1807  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1789 .. 1807  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1825 .. 1843  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
1825 .. 1843  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  1669 .. 2196  =  528 bp
ORF:  175 amino acids  =  19.8 kDa
ORF:  3521 .. 3853  =  333 bp
ORF:  110 amino acids  =  11.8 kDa
ORF:  2253 .. 3515  =  1263 bp
ORF:  420 amino acids  =  46.5 kDa
ORF:  6327 .. 6593  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  838 .. 1098  =  261 bp
ORF:  86 amino acids  =  9.5 kDa
ORF:  2418 .. 3248  =  831 bp
ORF:  276 amino acids  =  30.9 kDa
ORF:  4374 .. 4664  =  291 bp
ORF:  96 amino acids  =  10.5 kDa
ORF:  6197 .. 7057  =  861 bp
ORF:  286 amino acids  =  31.5 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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