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Plasmid Files

pTRE-Tight

Vector for expressing a gene with the Tet-On® Advanced or Tet-Off® Advanced system.

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pTRE-Tight Sequence and MappTRE-Tight.dna
Map and Sequence File   
Sequence Author:  Clontech
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 EcoO109I (2590) AatII (2536) ZraI (2534) SspI (2418) EarI (2409) XmnI (2213) ScaI (2094) TatI (2092) PvuI (1984) FspI (1836) AseI (1786) NmeAIII (1762) BglI (1734) BsrFI (1694) BsaI (1675) BmrI (1654) AhdI (1614) BbsI (2592) StuI (277) EcoRI (323) Acc65I (335) KpnI - TspMI - XmaI (339) SmaI (341) BamHI (344) PvuII (358) MluI (362) NheI (368) BmtI (372) EagI - NotI (375) BspDI - ClaI (383) HindIII (388) SalI (394) AccI (395) EcoRV (402) XbaI (406) BsaBI * (417) MfeI (505) BsmI (506) HpaI (518) PsiI (538) PciI (721) NspI (725) DrdI (829) HaeII (969) BseYI (1025) PspFI (1029) AlwNI (1137) pTRE-Tight 2605 bp
EcoO109I  (2590)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
AatII  (2536)
1 site
G A C G T C C T G C A G
ZraI  (2534)
1 site
G A C G T C C T G C A G
SspI  (2418)
1 site
A A T A T T T T A T A A
EarI  (2409)
1 site
C T C T T C N G A G A A G N N N N

Efficient cleavage requires at least two copies of the EarI
recognition sequence.
Sticky ends from different EarI sites may not be compatible.
XmnI  (2213)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2094)
1 site
A G T A C T T C A T G A
TatI  (2092)
1 site
W G T A C W W C A T G W
PvuI  (1984)
1 site
C G A T C G G C T A G C
FspI  (1836)
1 site
T G C G C A A C G C G T
AseI  (1786)
1 site
A T T A A T T A A T T A
NmeAIII  (1762)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII
recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BglI  (1734)
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.
BsrFI  (1694)
1 site
R C C G G Y Y G G C C R

Efficient cleavage requires at least two copies of the BsrFI
recognition sequence.
After cleavage, BsrFI can remain bound to DNA and alter its
electrophoretic mobility.
BsaI  (1675)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BmrI  (1654)
1 site
A C T G G G ( N ) 4 N T G A C C C ( N ) 4

The 1-base overhangs produced by BmrI may be hard to ligate.
Sticky ends from different BmrI sites may not be compatible.
Unlike most restriction enzymes, BmrI can cleave DNA in the
absence of magnesium.
AhdI  (1614)
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.
BbsI  (2592)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
StuI  (277)
1 site
A G G C C T T C C G G A
EcoRI  (323)
1 site
G A A T T C C T T A A G
Acc65I  (335)
1 site
G G T A C C C C A T G G
KpnI  (339)
1 site
G G T A C C C C A T G G
TspMI  (339)
1 site
C C C G G G G G G C C C
XmaI  (339)
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  (341)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (344)
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.
PvuII  (358)
1 site
C A G C T G G T C G A C
MluI  (362)
1 site
A C G C G T T G C G C A
NheI  (368)
1 site
G C T A G C C G A T C G
BmtI  (372)
1 site
G C T A G C C G A T C G
EagI  (375)
1 site
C G G C C G G C C G G C
NotI  (375)
1 site
G C G G C C G C C G C C G G C G
BspDI  (383)
1 site
A T C G A T T A G C T A
ClaI  (383)
1 site
A T C G A T T A G C T A
HindIII  (388)
1 site
A A G C T T T T C G A A
SalI  (394)
1 site
G T C G A C C A G C T G
AccI  (395)
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.
EcoRV  (402)
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.
XbaI  (406)
1 site
T C T A G A A G A T C T
BsaBI  (417)
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.
MfeI  (505)
1 site
C A A T T G G T T A A C
BsmI  (506)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
HpaI  (518)
1 site
G T T A A C C A A T T G
PsiI  (538)
1 site
T T A T A A A A T A T T
PciI  (721)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (725)
1 site
R C A T G Y Y G T A C R
DrdI  (829)
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.
HaeII  (969)
1 site
R G C G C Y Y C G C G R
BseYI  (1025)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its
electrophoretic mobility.
PspFI  (1029)
1 site
C C C A G C G G G T C G
AlwNI  (1137)
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.
AmpR
1541 .. 2401  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   1541 .. 2332  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1541 .. 2401  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   2333 .. 2401  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1541 .. 2401  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
782 .. 1370  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
782 .. 1370  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
tight TRE promoter
4 .. 318  =  315 bp
Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter
tight TRE promoter
4 .. 318  =  315 bp
Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter
AmpR promoter
2402 .. 2506  =  105 bp
AmpR promoter
2402 .. 2506  =  105 bp
MCS
323 .. 411  =  89 bp
multiple cloning site
MCS
323 .. 411  =  89 bp
multiple cloning site
SV40 poly(A) signal
519 .. 600  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
519 .. 600  =  82 bp
SV40 polyadenylation signal
tet operator
12 .. 30  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
12 .. 30  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
48 .. 66  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
48 .. 66  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
83 .. 101  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
83 .. 101  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
119 .. 137  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
119 .. 137  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
155 .. 173  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
155 .. 173  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
190 .. 208  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
190 .. 208  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
226 .. 244  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
226 .. 244  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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