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Plasmid Files

pTetOne™

All-in-one vector for strong inducible expression of genes in mammalian cells using the Tet-On® system.

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pTetOne Sequence and MappTetOne.dna
Map and Sequence File   
Sequence Author:  Clontech
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 NotI (4195) MluI (4188) AgeI (4176) BamHI (4170) BglII (4164) PstI (4162) AatII (3954) ZraI (3952) SspI (3836) XmnI (3631) PvuI (3402) AseI (3204) BglI (3152) AlwNI (2555) DrdI (2247) PciI (2139) EcoRI (4202) HindIII (114) BsgI (375) AarI (397) BstXI (548) StyI (566) PfoI (615) BsmBI - SpeI (801) MscI (867) BlpI (881) AscI (890) BspEI (995) StuI (1144) BsaAI - PmlI (1205) BseRI (1332) NruI (1354) TspMI - XmaI (1812) SmaI (1814) pTetOne™ 4207 bp
NotI  (4195)
1 site
G C G G C C G C C G C C G G C G
MluI  (4188)
1 site
A C G C G T T G C G C A
AgeI  (4176)
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.
BamHI  (4170)
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.
BglII  (4164)
1 site
A G A T C T T C T A G A
PstI  (4162)
1 site
C T G C A G G A C G T C
AatII  (3954)
1 site
G A C G T C C T G C A G
ZraI  (3952)
1 site
G A C G T C C T G C A G
SspI  (3836)
1 site
A A T A T T T T A T A A
XmnI  (3631)
1 site
G A A N N N N T T C C T T N N N N A A G
PvuI  (3402)
1 site
C G A T C G G C T A G C
AseI  (3204)
1 site
A T T A A T T A A T T A
BglI  (3152)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
AlwNI  (2555)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
DrdI  (2247)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PciI  (2139)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
EcoRI  (4202)
1 site
G A A T T C C T T A A G
HindIII  (114)
1 site
A A G C T T T T C G A A
BsgI  (375)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI
recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
AarI  (397)
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.
BstXI  (548)
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.
StyI  (566)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
PfoI  (615)
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.
BsmBI  (801)
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  (801)
1 site
A C T A G T T G A T C A
MscI  (867)
1 site
T G G C C A A C C G G T
BlpI  (881)
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.
AscI  (890)
1 site
G G C G C G C C C C G C G C G G
BspEI  (995)
1 site
T C C G G A A G G C C T
StuI  (1144)
1 site
A G G C C T T C C G G A
BsaAI  (1205)
1 site
Y A C G T R R T G C A Y
PmlI  (1205)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
BseRI  (1332)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
NruI  (1354)
1 site
T C G C G A A G C G C T
TspMI  (1812)
1 site
C C C G G G G G G C C C
XmaI  (1812)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI
recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (1814)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
AmpR
2959 .. 3819  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2959 .. 3750  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2959 .. 3819  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3751 .. 3819  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2959 .. 3819  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
Tet-On® 3G
1074 .. 1820  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
Tet-On® 3G
1074 .. 1820  =  747 bp
248 amino acids  =  27.7 kDa
Product: modified rtTA protein that binds tightly to
promoters containing the tet operator in the
presence of doxycycline
ori
2200 .. 2788  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2200 .. 2788  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
hPGK promoter
545 .. 1055  =  511 bp
human phosphoglycerate kinase 1 promoter
hPGK promoter
545 .. 1055  =  511 bp
human phosphoglycerate kinase 1 promoter
TRE3GS promoter
1 .. 368  =  368 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G, modified to
eliminate binding sites for endogenous mammalian
transcription factors
TRE3GS promoter
1 .. 368  =  368 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G, modified to
eliminate binding sites for endogenous mammalian
transcription factors
AmpR promoter
3820 .. 3924  =  105 bp
AmpR promoter
3820 .. 3924  =  105 bp
pause site
372 .. 463  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
pause site
372 .. 463  =  92 bp
RNA polymerase II transcriptional pause signal from
the human α2 globin gene
SV40 poly(A) signal
1937 .. 2018  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1937 .. 2018  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3957 .. 4038  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
3957 .. 4038  =  82 bp
SV40 polyadenylation signal
MCS
4158 .. 4207  =  50 bp
multiple cloning site
MCS
4158 .. 4207  =  50 bp
multiple cloning site
poly(A) signal
477 .. 525  =  49 bp
synthetic polyadenylation signal
poly(A) signal
477 .. 525  =  49 bp
synthetic polyadenylation signal
tet operator
126 .. 144  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
126 .. 144  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
162 .. 180  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
162 .. 180  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
198 .. 216  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
198 .. 216  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
234 .. 252  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
234 .. 252  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
270 .. 288  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
270 .. 288  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
306 .. 324  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
306 .. 324  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
342 .. 360  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
342 .. 360  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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