pRetroX-TetOne-Puro

All-in-one retroviral vector with a puromycin resistance marker, for strong inducible expression of genes using the Tet-On® system.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsViral Expression & Packaging VectorspRetroX-TetOne-Puro
BspQI - SapI (9222) PciI (9105) AlwNI (8696) ScaI (7736) SspI (7412) AleI (6074) EcoRV (5689) PflMI (5512) RsrII (4770) SnaBI (519) AfeI (717) Eco53kI (733) SacI (735) PshAI (1170) SrfI (1394) AflII (1416) Bsu36I (1428) Bpu10I (1700) PpuMI (2077) BamHI (2607) NgoMIV (2613) NaeI (2615) SgrAI (2619) MCS BstZ17I (2627) EcoRI (2633) HindIII (2752) PaeR7I - PspXI - XhoI (3004) hPGK promoter AscI (3368) BspEI (3473) NruI (3832) BfuAI - BspMI (4109) SfiI (4583) SalI (4762) pRetroX-TetOne™-Puro 9463 bp
BspQI  (9222)
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  (9222)
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.
PciI  (9105)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AlwNI  (8696)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
ScaI  (7736)
1 site		 A G T A C T T C A T G A
SspI  (7412)
1 site		 A A T A T T T T A T A A
AleI  (6074)
1 site		 C A C N N N N G T G G T G N N N N C A C
EcoRV  (5689)
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.
PflMI  (5512)
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.
RsrII  (4770)
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.
SnaBI  (519)
1 site		 T A C G T A A T G C A T
AfeI  (717)
1 site		 A G C G C T T C G C G A
Eco53kI  (733)
1 site		 G A G C T C C T C G A G
SacI  (735)
1 site		 G A G C T C C T C G A G
PshAI  (1170)
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.
SrfI  (1394)
1 site		 G C C C G G G C C G G G C C C G
AflII  (1416)
1 site		 C T T A A G G A A T T C
Bsu36I  (1428)
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.
Bpu10I  (1700)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
This recognition sequence is asymmetric, so ligating sticky ends generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
PpuMI  (2077)
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.
BamHI  (2607)
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.
NgoMIV  (2613)
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  (2615)
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.
SgrAI  (2619)
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.
BstZ17I  (2627)
1 site		 G T A T A C C A T A T G
EcoRI  (2633)
1 site		 G A A T T C C T T A A G
HindIII  (2752)
1 site		 A A G C T T T T C G A A
PaeR7I  (3004)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (3004)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (3004)
1 site		 C T C G A G G A G C T C
AscI  (3368)
1 site		 G G C G C G C C C C G C G C G G
BspEI  (3473)
1 site		 T C C G G A A G G C C T
NruI  (3832)
1 site		 T C G C G A A G C G C T
BfuAI  (4109)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (4109)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SfiI  (4583)
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.
SalI  (4762)
1 site		 G T C G A C C A G C T G
AmpR
7430 .. 8290  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7430 .. 7498  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7430 .. 8290  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7499 .. 8290  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7430 .. 8290  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
CMV/MMLV 5' LTR
164 .. 911  =  748 bp
chimera between the CMV enhancer/promoter and the 5' LTR of Moloney murine leukemia virus
CMV/MMLV 5' LTR
164 .. 911  =  748 bp
chimera between the CMV enhancer/promoter and the 5' LTR of Moloney murine leukemia virus
Tet-On® 3G
3552 .. 4298  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to promoters containing the tet operator in the presence of doxycycline
Tet-On® 3G
3552 .. 4298  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to promoters containing the tet operator in the presence of doxycycline
PuroR
4654 .. 5253  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
4654 .. 5253  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
ori
8461 .. 9049  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
8461 .. 9049  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hPGK promoter
3023 .. 3533  =  511 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
3023 .. 3533  =  511 bp
human phosphoglycerate kinase 1 promoter
WPRE
5271 .. 5674  =  404 bp
woodchuck posttranscriptional regulatory element
WPRE
5271 .. 5674  =  404 bp
woodchuck posttranscriptional regulatory element
TRE3GS promoter
2639 .. 3006  =  368 bp
3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On® 3G, modified to eliminate binding sites for endogenous mammalian transcription factors
TRE3GS promoter
2639 .. 3006  =  368 bp
3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On® 3G, modified to eliminate binding sites for endogenous mammalian transcription factors
MMLV Ψ
974 .. 1331  =  358 bp
packaging signal of Moloney murine leukemia virus (MMLV)
MMLV Ψ
974 .. 1331  =  358 bp
packaging signal of Moloney murine leukemia virus (MMLV)
SV40 promoter
4316 .. 4645  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4316 .. 4645  =  330 bp
SV40 enhancer and early promoter
3' LTR (ΔU3)
5835 .. 6047  =  213 bp
self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus
3' LTR (ΔU3)
5835 .. 6047  =  213 bp
self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus
SV40 poly(A) signal
2298 .. 2432  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2298 .. 2432  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6690 .. 6811  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6690 .. 6811  =  122 bp
SV40 polyadenylation signal
AmpR promoter
7325 .. 7429  =  105 bp
AmpR promoter
7325 .. 7429  =  105 bp
MCS
2607 .. 2638  =  32 bp
multiple cloning site
MCS
2607 .. 2638  =  32 bp
multiple cloning site
SV40 ori
4496 .. 4631  =  136 bp
SV40 origin of replication
SV40 ori
4496 .. 4631  =  136 bp
SV40 origin of replication
tet operator
2764 .. 2782  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2764 .. 2782  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2800 .. 2818  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2800 .. 2818  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2836 .. 2854  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2836 .. 2854  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2872 .. 2890  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2872 .. 2890  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2908 .. 2926  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2908 .. 2926  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2944 .. 2962  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2944 .. 2962  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2980 .. 2998  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
2980 .. 2998  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  1879 .. 2199  =  321 bp
ORF:  106 amino acids  =  11.8 kDa
ORF:  3709 .. 4011  =  303 bp
ORF:  100 amino acids  =  11.1 kDa
ORF:  4654 .. 5253  =  600 bp
ORF:  199 amino acids  =  21.5 kDa
ORF:  5374 .. 5670  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  6673 .. 7083  =  411 bp
ORF:  136 amino acids  =  15.7 kDa
ORF:  6143 .. 6460  =  318 bp
ORF:  105 amino acids  =  11.2 kDa
ORF:  7430 .. 8290  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3315 .. 4298  =  984 bp
ORF:  327 amino acids  =  36.0 kDa
ORF:  5361 .. 5759  =  399 bp
ORF:  132 amino acids  =  14.4 kDa
ORF:  9381 .. 160  =  243 bp
ORF:  80 amino acids  =  9.4 kDa
ORF:  4004 .. 4285  =  282 bp
ORF:  93 amino acids  =  9.6 kDa
ORF:  5873 .. 6511  =  639 bp
ORF:  212 amino acids  =  22.1 kDa
ORF:  7894 .. 8160  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2706 .. 2975  =  270 bp
ORF:  89 amino acids  =  10.5 kDa
ORF:  6765 .. 7043  =  279 bp
ORF:  92 amino acids  =  11.0 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

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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