pLenti3.3 TR

Lentiviral vector for constitutive high-level expression of the Tet repressor in the T-REx™ system.

Sequence Author: Thermo Fisher (Invitrogen)

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HomePlasmidsViral Expression & Packaging VectorspLenti3.3 TR
PciI (8818) AhdI (7930) DraIII (6790) SfiI (6302) RsrII (5527) PflFI - Tth111I (5129) MscI (5093) BsaBI * (4870) CsiI - SexAI * (4568) NotI (920) BbvCI (1196) HpaI (1800) cPPT/CTS BspDI - ClaI (1923) BsmBI - Esp3I (2472) SpeI (2534) BstXI (2547) EcoRV (2563) PspOMI (3094) ApaI (3098) AfeI (3627) MluI (3871) PflMI (4138) PfoI (4170) SacII (4400) pLenti3.3/TR 9228 bp
PciI  (8818)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
AhdI  (7930)
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.
DraIII  (6790)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
SfiI  (6302)
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.
RsrII  (5527)
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.
PflFI  (5129)
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  (5129)
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.
MscI  (5093)
1 site		 T G G C C A A C C G G T
BsaBI  (4870)
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.
CsiI  (4568)
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  (4568)
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.
NotI  (920)
1 site		 G C G G C C G C C G C C G G C G
BbvCI  (1196)
1 site		 C C T C A G C G G A G T C G
HpaI  (1800)
1 site		 G T T A A C C A A T T G
BspDI  (1923)
1 site		 A T C G A T T A G C T A
ClaI  (1923)
1 site		 A T C G A T T A G C T A
BsmBI  (2472)
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  (2472)
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.
SpeI  (2534)
1 site		 A C T A G T T G A T C A
BstXI  (2547)
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.
EcoRV  (2563)
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.
PspOMI  (3094)
1 site		 G G G C C C C C C G G G
ApaI  (3098)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
AfeI  (3627)
1 site		 A G C G C T T C G C G A
MluI  (3871)
1 site		 A C G C G T T G C G C A
PflMI  (4138)
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.
PfoI  (4170)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
SacII  (4400)
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.
AmpR
7143 .. 8003  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7143 .. 7211  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7143 .. 8003  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7212 .. 8003  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7143 .. 8003  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
NeoR/KanR
4883 .. 5677  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
NeoR/KanR
4883 .. 5677  =  795 bp
264 amino acids  =  29.0 kDa
Product: aminoglycoside phosphotransferase from Tn5
confers resistance to neomycin, kanamycin, and G418 (Geneticin)
TetR
3223 .. 3840  =  618 bp
206 amino acids  =  23.3 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
TetR
3223 .. 3840  =  618 bp
206 amino acids  =  23.3 kDa
Product: tetracycline repressor TetR
TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline.
WPRE
3897 .. 4485  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
WPRE
3897 .. 4485  =  589 bp
woodchuck hepatitis virus posttranscriptional regulatory element
ori
8174 .. 8762  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
8174 .. 8762  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
β-globin intron
2567 .. 3139  =  573 bp
intron from rabbit β-globin gene
β-globin intron
2567 .. 3139  =  573 bp
intron from rabbit β-globin gene
f1 ori
6557 .. 7012  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6557 .. 7012  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
SV40 promoter
4500 .. 4816  =  317 bp
SV40 enhancer and early promoter
SV40 promoter
4500 .. 4816  =  317 bp
SV40 enhancer and early promoter
CMV enhancer
1944 .. 2247  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1944 .. 2247  =  304 bp
human cytomegalovirus immediate early enhancer
RRE
1075 .. 1308  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
RRE
1075 .. 1308  =  234 bp
The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm.
3' LTR (ΔU3)
5749 .. 5982  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
3' LTR (ΔU3)
5749 .. 5982  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from HIV-1
RSV promoter
1 .. 229  =  229 bp
Rous sarcoma virus enhancer/promoter
RSV promoter
1 .. 229  =  229 bp
Rous sarcoma virus enhancer/promoter
CMV promoter
2248 .. 2451  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
CMV promoter
2248 .. 2451  =  204 bp
human cytomegalovirus (CMV) immediate early promoter
5' LTR (truncated)
230 .. 410  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
5' LTR (truncated)
230 .. 410  =  181 bp
truncated 5' long terminal repeat (LTR) from HIV-1
HIV-1 Ψ
457 .. 582  =  126 bp
packaging signal of human immunodeficiency virus type 1
HIV-1 Ψ
457 .. 582  =  126 bp
packaging signal of human immunodeficiency virus type 1
SV40 poly(A) signal
6054 .. 6175  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
6054 .. 6175  =  122 bp
SV40 polyadenylation signal
cPPT/CTS
1804 .. 1921  =  118 bp
central polypurine tract and central termination sequence of HIV-1
cPPT/CTS
1804 .. 1921  =  118 bp
central polypurine tract and central termination sequence of HIV-1
AmpR promoter
7038 .. 7142  =  105 bp
AmpR promoter
7038 .. 7142  =  105 bp
SV40 ori
4667 .. 4802  =  136 bp
SV40 origin of replication
SV40 ori
4667 .. 4802  =  136 bp
SV40 origin of replication
ORF:  3223 .. 3870  =  648 bp
ORF:  215 amino acids  =  24.3 kDa
ORF:  4000 .. 4296  =  297 bp
ORF:  98 amino acids  =  10.2 kDa
ORF:  4492 .. 4764  =  273 bp
ORF:  90 amino acids  =  10.3 kDa
ORF:  6076 .. 6312  =  237 bp
ORF:  78 amino acids  =  8.6 kDa
ORF:  9160 .. 177  =  246 bp
ORF:  81 amino acids  =  8.9 kDa
ORF:  953 .. 1963  =  1011 bp
ORF:  336 amino acids  =  38.5 kDa
ORF:  4307 .. 4561  =  255 bp
ORF:  84 amino acids  =  8.9 kDa
ORF:  4883 .. 5677  =  795 bp
ORF:  264 amino acids  =  29.0 kDa
ORF:  3888 .. 4646  =  759 bp
ORF:  252 amino acids  =  27.2 kDa
ORF:  5055 .. 5441  =  387 bp
ORF:  128 amino acids  =  14.6 kDa
ORF:  7143 .. 8003  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  868 .. 1248  =  381 bp
ORF:  126 amino acids  =  13.3 kDa
ORF:  9087 .. 158  =  300 bp
ORF:  99 amino acids  =  10.9 kDa
ORF:  3159 .. 3392  =  234 bp
ORF:  77 amino acids  =  8.4 kDa
ORF:  5192 .. 5446  =  255 bp
ORF:  84 amino acids  =  9.6 kDa
ORF:  7607 .. 7873  =  267 bp
ORF:  88 amino acids  =  9.2 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