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Plasmid Files

pLVX-Tet-On Advanced

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

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pLVX-Tet-On Advanced Sequence and MappLVX-Tet-On Advanced.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SgrDI (11,258) SspI (11,141) AhdI (10,337) BstZ17I (9065) AsiSI (8142) PshAI (7805) 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) AarI (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.
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

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
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.
AarI  (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 AarI
recognition sequence.
Sticky ends from different AarI sites may not be compatible.
After cleavage, AarI can remain bound to DNA and alter its
electrophoretic mobility.
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
   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
   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
   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
   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
   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
   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
   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
   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
   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
   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
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