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Plasmid Files

pTRE3G-BI-mCherry

Vector for doxycycline-inducible expression of a gene together with the mCherry fluorescent protein.

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pTRE3G-BI-mCherry Sequence and MappTRE3G-BI-mCherry.dna
Map and Sequence File   
Sequence Author:  Clontech
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 EcoRI (3554) MscI (3519) BsaAI (3319) PstI - SbfI (3198) PflMI (3113) BbsI (3110) BbvCI - Bpu10I (3008) PvuII (2971) BsgI (2957) XcmI (2864) BsrGI (2845) XbaI (2834) AatII (2642) ZraI (2640) SspI (2524) XmnI (2319) ScaI (2200) PvuI (2090) FspI (1942) AseI (1892) BglI (1840) BsaI (1781) PaeR7I - PspXI - XhoI (70) HindIII (322) SalI (452) AccI (453) EcoO109I - PspOMI (462) ApaI (466) BglII (468) BamHI (480) BspDI * - ClaI * (487) EagI - NotI (495) MreI - NgoMIV (500) NaeI (502) EcoRV (508) AflIII - PciI (827) NspI (831) BmrI (1760) pTRE3G-BI-mCherry 3564 bp
EcoRI  (3554)
1 site
G A A T T C C T T A A G
MscI  (3519)
1 site
T G G C C A A C C G G T
BsaAI  (3319)
1 site
Y A C G T R R T G C A Y
PstI  (3198)
1 site
C T G C A G G A C G T C
SbfI  (3198)
1 site
C C T G C A G G G G A C G T C C
PflMI  (3113)
1 site
C C A N N N N N T G G G G T N N N N N A C C

Sticky ends from different PflMI sites may not be compatible.
BbsI  (3110)
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.
BbvCI  (3008)
1 site
C C T C A G C G G A G T C G
Bpu10I  (3008)
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.
PvuII  (2971)
1 site
C A G C T G G T C G A C
BsgI  (2957)
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).
XcmI  (2864)
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.
BsrGI  (2845)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
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
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
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.
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
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.
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.
AflIII  (827)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
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
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.
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
mCherry
2840 .. 3550  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
mCherry
2840 .. 3550  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
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
452 .. 511  =  60 bp
multiple cloning site
MCS
452 .. 511  =  60 bp
multiple cloning site
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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