ptTS-Neo

Vector for expressing tTS, which represses expression from Tet-responsive promoters in the absence of doxycycline.

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsViral Expression & Packaging VectorsptTS-Neo
EcoNI (4) XmnI (5977) PvuI (5748) BsaI (5439) AhdI (5378) Bpu10I (4182) HindIII (4159) PflMI (3912) BglII (3705) Bsu36I (3635) ApaI (3535) PspOMI (3531) PsiI (3364) MfeI (3356) NsiI (61) SfiI (192) StuI (238) BsaBI * (276) EagI (323) KasI (416) NarI (417) SfoI (418) PluTI (420) PflFI - Tth111I (535) RsrII (933) BstBI (1099) BsrGI (1407) SpeI (1463) SnaBI (1804) Eco53kI (2030) SacI (2032) BsmBI - Esp3I (2053) BbsI (2098) SacII (2119) XbaI (2138) PmlI (2266) NheI (2785) BmtI (2789) AvaI - BsoBI (2829) BamHI (2971) KflI (3014) ptTS-Neo 6365 bp
EcoNI  (4)
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.
XmnI  (5977)
1 site		 G A A N N N N T T C C T T N N N N A A G
PvuI  (5748)
1 site		 C G A T C G G C T A G C
BsaI  (5439)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AhdI  (5378)
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.
Bpu10I  (4182)
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.
HindIII  (4159)
1 site		 A A G C T T T T C G A A
PflMI  (3912)
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.
BglII  (3705)
1 site		 A G A T C T T C T A G A
Bsu36I  (3635)
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.
ApaI  (3535)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (3531)
1 site		 G G G C C C C C C G G G
PsiI  (3364)
1 site		 T T A T A A A A T A T T
MfeI  (3356)
1 site		 C A A T T G G T T A A C
NsiI  (61)
1 site		 A T G C A T T A C G T A
SfiI  (192)
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.
StuI  (238)
1 site		 A G G C C T T C C G G A
BsaBI  (276)
1 site		 G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
EagI  (323)
1 site		 C G G C C G G C C G G C
KasI  (416)
1 site		 G G C G C C C C G C G G
NarI  (417)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (418)
1 site		 G G C G C C C C G C G G
PluTI  (420)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
PflFI  (535)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (535)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
RsrII  (933)
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.
BstBI  (1099)
1 site		 T T C G A A A A G C T T
BsrGI  (1407)
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.
SpeI  (1463)
1 site		 A C T A G T T G A T C A
SnaBI  (1804)
1 site		 T A C G T A A T G C A T
Eco53kI  (2030)
1 site		 G A G C T C C T C G A G
SacI  (2032)
1 site		 G A G C T C C T C G A G
BsmBI  (2053)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2053)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
BbsI  (2098)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
SacII  (2119)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
XbaI  (2138)
1 site		 T C T A G A A G A T C T
PmlI  (2266)
1 site		 C A C G T G G T G C A C
NheI  (2785)
1 site		 G C T A G C C G A T C G
BmtI  (2789)
1 site		 G C T A G C C G A T C G
AvaI  (2829)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different AvaI sites may not be compatible.
BsoBI  (2829)
1 site		 C Y C G R G G R G C Y C
Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
BamHI  (2971)
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.
KflI  (3014)
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.
AmpR
5305 .. 6165  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 2:  
   5305 .. 6096  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5305 .. 6165  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
   Segment 1:  signal sequence  
   6097 .. 6165  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5305 .. 6165  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
tTS
2135 .. 2983  =  849 bp
282 amino acids  =  31.9 kDa
3 segments
   Segment 1:  TetR  
   2135 .. 2767  =  633 bp
   211 amino acids  =  23.6 kDa
Product: fusion of a hybrid tetracycline repressor TetR with a nuclear localization signal and a KRAB silencing domain
silences Tet-responsive promoters in the absence but not the presence of doxycycline
tTS
2135 .. 2983  =  849 bp
282 amino acids  =  31.9 kDa
3 segments
   Segment 2:  NLS  
   2768 .. 2788  =  21 bp
   7 amino acids  =  897.2 Da
Product: fusion of a hybrid tetracycline repressor TetR with a nuclear localization signal and a KRAB silencing domain
silences Tet-responsive promoters in the absence but not the presence of doxycycline
tTS
2135 .. 2983  =  849 bp
282 amino acids  =  31.9 kDa
3 segments
   Segment 3:  KRAB silencing domain  
   2789 .. 2983  =  195 bp
   64 amino acids  =  7.4 kDa
Product: fusion of a hybrid tetracycline repressor TetR with a nuclear localization signal and a KRAB silencing domain
silences Tet-responsive promoters in the absence but not the presence of doxycycline
tTS
2135 .. 2983  =  849 bp
282 amino acids  =  31.9 kDa
3 segments
Product: fusion of a hybrid tetracycline repressor TetR with a nuclear localization signal and a KRAB silencing domain
silences Tet-responsive promoters in the absence but not the presence of doxycycline
NeoR/KanR
289 .. 1083  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
289 .. 1083  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
ori
4546 .. 5134  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
4546 .. 5134  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
β-globin intron
3004 .. 3576  =  573 bp
intron from rabbit β-globin gene
β-globin intron
3004 .. 3576  =  573 bp
intron from rabbit β-globin gene
CMV enhancer
1449 .. 1828  =  380 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1449 .. 1828  =  380 bp
human cytomegalovirus immediate early enhancer
CMV promoter
1829 .. 2032  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
1829 .. 2032  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
SV40 promoter
58 .. 254  =  197 bp
SV40 early promoter
SV40 promoter
58 .. 254  =  197 bp
SV40 early promoter
AmpR promoter
6166 .. 6270  =  105 bp
AmpR promoter
6166 .. 6270  =  105 bp
β-globin poly(A) signal
3773 .. 3828  =  56 bp
rabbit β-globin polyadenylation signal
β-globin poly(A) signal
3773 .. 3828  =  56 bp
rabbit β-globin polyadenylation signal
HSV TK poly(A) signal
1315 .. 1363  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
HSV TK poly(A) signal
1315 .. 1363  =  49 bp
herpesvirus thymidine kinase polyadenylation signal
SV40 ori
105 .. 240  =  136 bp
SV40 origin of replication
SV40 ori
105 .. 240  =  136 bp
SV40 origin of replication
ORF:  289 .. 1083  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  461 .. 847  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  1988 .. 2983  =  996 bp
ORF:  331 amino acids  =  38.0 kDa
ORF:  5435 .. 5701  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  1104 .. 1463  =  360 bp
ORF:  119 amino acids  =  13.7 kDa
ORF:  4146 .. 4445  =  300 bp
ORF:  99 amino acids  =  11.4 kDa
ORF:  1163 .. 1402  =  240 bp
ORF:  79 amino acids  =  9.0 kDa
ORF:  598 .. 1134  =  537 bp
ORF:  178 amino acids  =  19.8 kDa
ORF:  2047 .. 2295  =  249 bp
ORF:  82 amino acids  =  9.4 kDa
ORF:  5305 .. 6165  =  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

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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