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Plasmid Files

pLenti3.3/TR

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

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pLenti3.3 TR Sequence and MappLenti3.3 TR.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 PciI (8818) AhdI (7930) DraIII (6790) SfiI (6302) RsrII (5527) PflFI - Tth111I (5129) MscI (5093) BsaBI * (4870) NotI (920) BbvCI (1196) HpaI (1800) BspDI - ClaI (1923) BsmBI (2472) SpeI (2534) BstXI (2547) EcoRV (2563) PspOMI (3094) ApaI (3098) AfeI (3627) MluI (3871) PflMI (4138) PfoI (4170) SacII (4400) SexAI * (4568) 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.
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.
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.
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.
AmpR
7143 .. 8003  =  861 bp
286 amino acids  =  31.6 kDa
   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
   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
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