pLVX-Tet-On Advanced

Lentiviral vector for producing a doxycycline-activated transactivator in a Tet-On system.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsViral Expression & Packaging VectorspLVX-Tet-On Advanced
SgrDI (11,258) SspI (11,141) AhdI (10,337) BstZ17I (9065) AsiSI (8142) PshAI (7805) CsiI - SexAI * (7481) PmeI (6666) BmtI (6661) NheI (6657) EcoNI (1170) BbvCI - Bpu10I (1424) AleI (1577) KflI - PpuMI (1934) cPPT/CTS BspDI - ClaI (2180) SnaBI (2480) BamHI (2788) XcmI (2802) PaqCI (3909) BmgBI (4113) BstXI (4156) MscI (4367) AfeI (5074) PaeR7I - PspXI - XhoI (5115) MluI (5134) pLVX-Tet-On Advanced 11,625 bp
SgrDI  (11,258)
1 site		 C G T C G A C G G C A G C T G C
SspI  (11,141)
1 site		 A A T A T T T T A T A A
AhdI  (10,337)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BstZ17I  (9065)
1 site		 G T A T A C C A T A T G
AsiSI  (8142)
1 site		 G C G A T C G C C G C T A G C G
PshAI  (7805)
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.
CsiI  (7481)
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  (7481)
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.
PmeI  (6666)
1 site		 G T T T A A A C C A A A T T T G
BmtI  (6661)
1 site		 G C T A G C C G A T C G
NheI  (6657)
1 site		 G C T A G C C G A T C G
EcoNI  (1170)
1 site		 C C T N N N N N A G G G G A N N N N N T C C
The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BbvCI  (1424)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (1424)
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.
AleI  (1577)
1 site		 C A C N N N N G T G G T G N N N N C A C
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
SnaBI  (2480)
1 site		 T A C G T A A T G C A T
BamHI  (2788)
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.
XcmI  (2802)
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.
PaqCI  (3909)
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  (4113)
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.
BstXI  (4156)
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.
MscI  (4367)
1 site		 T G G C C A A C C G G T
AfeI  (5074)
1 site		 A G C G C T T C G C G A
PaeR7I  (5115)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5115)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (5115)
1 site		 C T C G A G G A G C T C
MluI  (5134)
1 site		 A C G C G T T G C G C A
HygR
7778 .. 8803  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
HygR
7778 .. 8803  =  1026 bp
341 amino acids  =  38.0 kDa
Product: aminoglycoside phosphotransferase from E. coli
confers resistance to hygromycin
AmpR
10,264 .. 11,124  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   10,264 .. 11,055  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,264 .. 11,124  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   11,056 .. 11,124  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
10,264 .. 11,124  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
4157 .. 4951  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
4157 .. 4951  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 1:  rTetR  
   2797 .. 3414  =  618 bp
   206 amino acids  =  23.2 kDa
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 2:  
   3415 .. 3420  =  6 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 3:  VP16 minimal transcriptional activation domain  
   3421 .. 3456  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 4:  
   3457 .. 3459  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 5:  VP16 minimal transcriptional activation domain  
   3460 .. 3495  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 6:  
   3496 .. 3498  =  3 bp
   1 amino acid  =  115.1 Da
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 7:  VP16 minimal transcriptional activation domain  
   3499 .. 3534  =  36 bp
   12 amino acids  =  1.4 kDa
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
   Segment 8:  
   3535 .. 3543  =  9 bp
   2 amino acids  =  172.2 Da
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
Tet-On Advanced
2797 .. 3543  =  747 bp
248 amino acids  =  27.7 kDa
8 segments
Product: improved tetracycline-controlled transactivator
binds to a Tet-responsive element in the presence but not the absence of doxycycline
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
5' LTR
1 .. 634  =  634 bp
5' long terminal repeat (LTR) from HIV-1
3' LTR
5942 .. 6575  =  634 bp
3' long terminal repeat (LTR) from HIV-1
3' LTR
5942 .. 6575  =  634 bp
3' long terminal repeat (LTR) from HIV-1
WPRE
5147 .. 5735  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
5147 .. 5735  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
9505 .. 10,093  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
9505 .. 10,093  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
IRES
3570 .. 4143  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
IRES
3570 .. 4143  =  574 bp
internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)
f1 ori
6958 .. 7386  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6958 .. 7386  =  429 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
7400 .. 7729  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
7400 .. 7729  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
2201 .. 2504  =  304 bp
human cytomegalovirus immediate early enhancer
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.
bGH poly(A) signal
6688 .. 6912  =  225 bp
bovine growth hormone polyadenylation signal
bGH poly(A) signal
6688 .. 6912  =  225 bp
bovine growth hormone polyadenylation signal
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2505 .. 2708  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 poly(A) signal
11,277 .. 11,411  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
11,277 .. 11,411  =  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
SV40 poly(A) signal
8933 .. 9054  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
8933 .. 9054  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
cPPT/CTS
2028 .. 2144  =  117 bp
central polypurine tract and central termination sequence of HIV-1 (lacking the first T)
AmpR promoter
11,125 .. 11,229  =  105 bp
AmpR promoter
11,125 .. 11,229  =  105 bp
SV40 ori
7580 .. 7715  =  136 bp
SV40 origin of replication
SV40 ori
7580 .. 7715  =  136 bp
SV40 origin of replication
ORF:  2797 .. 3543  =  747 bp
ORF:  248 amino acids  =  27.7 kDa
ORF:  1181 .. 2032  =  852 bp
ORF:  283 amino acids  =  32.7 kDa
ORF:  2954 .. 3256  =  303 bp
ORF:  100 amino acids  =  11.1 kDa
ORF:  4157 .. 4951  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  5237 .. 5764  =  528 bp
ORF:  175 amino acids  =  19.0 kDa
ORF:  7778 .. 8803  =  1026 bp
ORF:  341 amino acids  =  38.0 kDa
ORF:  10,394 .. 10,660  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  4053 .. 4328  =  276 bp
ORF:  91 amino acids  =  10.3 kDa
ORF:  4329 .. 4715  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  5250 .. 5546  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  6909 .. 7175  =  267 bp
ORF:  88 amino acids  =  9.3 kDa
ORF:  1015 .. 1476  =  462 bp
ORF:  153 amino acids  =  16.6 kDa
ORF:  10,264 .. 11,124  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3249 .. 3530  =  282 bp
ORF:  93 amino acids  =  9.5 kDa
ORF:  4466 .. 4720  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  7715 .. 8536  =  822 bp
ORF:  273 amino acids  =  30.8 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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