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Plasmid Files

pLenti6/TR

Lentiviral vector for high-level constitutive expression of the Tet repressor.

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pLenti6 TR Sequence and MappLenti6 TR.dna
Map and Sequence File   
Sequence Author:  Invitrogen (Life Technologies)
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 KasI (413) PciI (7944) AhdI (7056) DraIII (5916) NaeI (5810) NgoMIV (5808) SfiI (5428) BlpI (4785) BsaBI * (4688) MscI (4393) PmlI (4321) NarI (414) SfoI (415) PluTI (417) AccI (738) NotI (920) BbvCI (1196) BsmBI (2344) SpeI (2406) BstXI (2419) PstI (2432) BspEI * (2526) AfeI (3612) SacII (3864) BstBI (3868) AgeI (3909) SexAI * (4028) pLenti6/TR 8354 bp
KasI  (413)
1 site
G G C G C C C C G C G G
PciI  (7944)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AhdI  (7056)
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  (5916)
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.
NaeI  (5810)
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.
NgoMIV  (5808)
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.
SfiI  (5428)
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.
BlpI  (4785)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BsaBI  (4688)
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.
MscI  (4393)
1 site
T G G C C A A C C G G T
PmlI  (4321)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
NarI  (414)
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  (415)
1 site
G G C G C C C C G C G G
PluTI  (417)
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.
AccI  (738)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the
recognition sequence.
Sticky ends from different AccI 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
BsmBI  (2344)
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  (2406)
1 site
A C T A G T T G A T C A
BstXI  (2419)
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.
PstI  (2432)
1 site
C T G C A G G A C G T C
BspEI  (2526)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
AfeI  (3612)
1 site
A G C G C T T C G C G A
SacII  (3864)
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.
BstBI  (3868)
1 site
T T C G A A A A G C T T
AgeI  (3909)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
SexAI  (4028)
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
6269 .. 7129  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   6269 .. 6337  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6269 .. 7129  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   6338 .. 7129  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
6269 .. 7129  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
TetR
3208 .. 3828  =  621 bp
207 amino acids  =  23.4 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
3208 .. 3828  =  621 bp
207 amino acids  =  23.4 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.
ori
7300 .. 7888  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
7300 .. 7888  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
β-globin intron
2552 .. 3124  =  573 bp
intron from rabbit β-globin gene
β-globin intron
2552 .. 3124  =  573 bp
intron from rabbit β-globin gene
f1 ori
5683 .. 6138  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
5683 .. 6138  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
BSD
4390 .. 4788  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
BSD
4390 .. 4788  =  399 bp
132 amino acids  =  13.7 kDa
Product: blasticidin S deaminase
confers resistance to blasticidin
SV40 promoter
3947 .. 4276  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
3947 .. 4276  =  330 bp
SV40 enhancer and early promoter
CMV enhancer
1816 .. 2119  =  304 bp
human cytomegalovirus immediate early enhancer
CMV enhancer
1816 .. 2119  =  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)
4875 .. 5108  =  234 bp
self-inactivating 3' long terminal repeat (LTR) from
HIV-1
3' LTR (ΔU3)
4875 .. 5108  =  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
2120 .. 2323  =  204 bp
human cytomegalovirus (CMV) immediate early
promoter
CMV promoter
2120 .. 2323  =  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
5180 .. 5301  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
5180 .. 5301  =  122 bp
SV40 polyadenylation signal
AmpR promoter
6164 .. 6268  =  105 bp
AmpR promoter
6164 .. 6268  =  105 bp
EM7 promoter
4324 .. 4371  =  48 bp
synthetic bacterial promoter
EM7 promoter
4324 .. 4371  =  48 bp
synthetic bacterial promoter
SV40 ori
4127 .. 4262  =  136 bp
SV40 origin of replication
SV40 ori
4127 .. 4262  =  136 bp
SV40 origin of replication
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