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Plasmid Files

pTRE3G-BI

Vector for doxycycline-inducible expression of two genes in cells containing the Tet-On® 3G transactivator protein.

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pTRE3G-BI Sequence and MappTRE3G-BI.dna
Map and Sequence File   
Sequence Author:  Clontech
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 SmaI (2866) TspMI - XmaI (2864) KpnI (2862) Acc65I (2858) MluI (2852) PstI (2850) NdeI (2841) XbaI (2834) AatII (2642) ZraI (2640) SspI (2524) XmnI (2319) ScaI (2200) TatI (2198) PvuI (2090) FspI (1942) AseI (1892) NmeAIII (1868) BglI (1840) BsaI (1781) BmrI (1760) AhdI (1720) EcoRI (2870) StuI (40) PaeR7I - PspXI - XhoI (70) HindIII (322) KasI (343) NarI (344) SfoI (345) PluTI (347) SalI (452) AccI (453) EcoO109I - PspOMI (462) ApaI (466) BglII (468) BamHI (480) BspDI * - ClaI * (487) EagI - NotI (495) MreI - NgoMIV - SgrAI (500) NaeI (502) EcoRV (508) PciI (827) NspI (831) DrdI (935) BseYI (1131) PspFI (1135) AlwNI (1243) pTRE3G-BI 2880 bp
SmaI  (2866)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (2864)
1 site
C C C G G G G G G C C C
XmaI  (2864)
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.
KpnI  (2862)
1 site
G G T A C C C C A T G G
Acc65I  (2858)
1 site
G G T A C C C C A T G G
MluI  (2852)
1 site
A C G C G T T G C G C A
PstI  (2850)
1 site
C T G C A G G A C G T C
NdeI  (2841)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional
nucleotides.
XbaI  (2834)
1 site
T C T A G A A G A T C T
AatII  (2642)
1 site
G A C G T C C T G C A G
ZraI  (2640)
1 site
G A C G T C C T G C A G
SspI  (2524)
1 site
A A T A T T T T A T A A
XmnI  (2319)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2200)
1 site
A G T A C T T C A T G A
TatI  (2198)
1 site
W G T A C W W C A T G W
PvuI  (2090)
1 site
C G A T C G G C T A G C
FspI  (1942)
1 site
T G C G C A A C G C G T
AseI  (1892)
1 site
A T T A A T T A A T T A
NmeAIII  (1868)
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  (1840)
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.
BsaI  (1781)
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  (1760)
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  (1720)
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.
EcoRI  (2870)
1 site
G A A T T C C T T A A G
StuI  (40)
1 site
A G G C C T T C C G G A
PaeR7I  (70)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (70)
1 site
V C T C G A G B B G A G C T C V
XhoI  (70)
1 site
C T C G A G G A G C T C
HindIII  (322)
1 site
A A G C T T T T C G A A
KasI  (343)
1 site
G G C G C C C C G C G G
NarI  (344)
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  (345)
1 site
G G C G C C C C G C G G
PluTI  (347)
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.
SalI  (452)
1 site
G T C G A C C A G C T G
AccI  (453)
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.
EcoO109I  (462)
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.
PspOMI  (462)
1 site
G G G C C C C C C G G G
ApaI  (466)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
BglII  (468)
1 site
A G A T C T T C T A G A
BamHI  (480)
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.
BspDI  (487)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (487)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EagI  (495)
1 site
C G G C C G G C C G G C
NotI  (495)
1 site
G C G G C C G C C G C C G G C G
MreI  (500)
1 site
C G C C G G C G G C G G C C G C
NgoMIV  (500)
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.
SgrAI  (500)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
NaeI  (502)
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.
EcoRV  (508)
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.
PciI  (827)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
NspI  (831)
1 site
R C A T G Y Y G T A C R
DrdI  (935)
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.
BseYI  (1131)
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  (1135)
1 site
C C C A G C G G G T C G
AlwNI  (1243)
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
1647 .. 2507  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1647 .. 2438  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1647 .. 2507  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2439 .. 2507  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1647 .. 2507  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
888 .. 1476  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
888 .. 1476  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
TRE3G BI promoter
1 .. 451  =  451 bp
3rd-generation Tet-responsive bidirectional
promoter that can be activated by binding of Tet-On
3G
TRE3G BI promoter
1 .. 451  =  451 bp
3rd-generation Tet-responsive bidirectional
promoter that can be activated by binding of Tet-On
3G
AmpR promoter
2508 .. 2612  =  105 bp
AmpR promoter
2508 .. 2612  =  105 bp
SV40 poly(A) signal
625 .. 706  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
625 .. 706  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2645 .. 2726  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2645 .. 2726  =  82 bp
SV40 polyadenylation signal
MCS 1
452 .. 511  =  60 bp
multiple cloning site 1
MCS 1
452 .. 511  =  60 bp
multiple cloning site 1
MCS 2
2834 .. 2875  =  42 bp
multiple cloning site 2
MCS 2
2834 .. 2875  =  42 bp
multiple cloning site 2
tet operator
81 .. 99  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
81 .. 99  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
117 .. 135  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
117 .. 135  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
153 .. 171  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
153 .. 171  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
189 .. 207  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
189 .. 207  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
225 .. 243  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
225 .. 243  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
261 .. 279  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
261 .. 279  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
297 .. 315  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
297 .. 315  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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