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Plasmid Files

pTRE-Dual2

Vector for co-expressing mCherry and another gene with the Tet-On® Advanced or Tet-Off® Advanced system.

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pTRE-Dual2 Sequence and MappTRE-Dual2.dna
Map and Sequence File   
Sequence Author:  Clontech
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 ZraI (3813) SspI (3697) EarI (3688) ScaI (3373) PvuI (3263) FspI (3115) AseI (3065) BsaI (2954) AatII (3815) Eco53kI (292) SacI (294) EcoRI (323) PstI - SbfI (690) BbvCI - Bpu10I (873) PvuII (913) BsgI (929) SgrAI (1016) XcmI (1021) BsrGI (1035) NheI (1046) BmtI (1050) PspOMI (1166) ApaI (1170) AvrII (1204) HindIII (1277) PmlI (1369) AarI - BfuAI - BspMI (1392) DraIII (1416) Acc65I (1494) KpnI (1498) BmgBI (1596) EagI - NotI - SacII (1641) BglII (1649) BamHI (1655) BspDI - ClaI (1662) SalI (1667) AccI (1668) EcoRV (1675) NdeI (1680) XbaI (1685) MfeI (1784) HpaI (1797) PsiI (1817) pTRE-Dual2 3884 bp
ZraI  (3813)
1 site
G A C G T C C T G C A G
SspI  (3697)
1 site
A A T A T T T T A T A A
EarI  (3688)
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.
ScaI  (3373)
1 site
A G T A C T T C A T G A
PvuI  (3263)
1 site
C G A T C G G C T A G C
FspI  (3115)
1 site
T G C G C A A C G C G T
AseI  (3065)
1 site
A T T A A T T A A T T A
BsaI  (2954)
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.
AatII  (3815)
1 site
G A C G T C C T G C A G
Eco53kI  (292)
1 site
G A G C T C C T C G A G
SacI  (294)
1 site
G A G C T C C T C G A G
EcoRI  (323)
1 site
G A A T T C C T T A A G
PstI  (690)
1 site
C T G C A G G A C G T C
SbfI  (690)
1 site
C C T G C A G G G G A C G T C C
BbvCI  (873)
1 site
C C T C A G C G G A G T C G
Bpu10I  (873)
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  (913)
1 site
C A G C T G G T C G A C
BsgI  (929)
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).
SgrAI  (1016)
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.
XcmI  (1021)
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  (1035)
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.
NheI  (1046)
1 site
G C T A G C C G A T C G
BmtI  (1050)
1 site
G C T A G C C G A T C G
PspOMI  (1166)
1 site
G G G C C C C C C G G G
ApaI  (1170)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (1204)
1 site
C C T A G G G G A T C C
HindIII  (1277)
1 site
A A G C T T T T C G A A
PmlI  (1369)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (1392)
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.
BfuAI  (1392)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BfuAI
recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (1392)
1 site
A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4

Efficient cleavage requires at least two copies of the BspMI
recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
DraIII  (1416)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
Acc65I  (1494)
1 site
G G T A C C C C A T G G
KpnI  (1498)
1 site
G G T A C C C C A T G G
BmgBI  (1596)
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.
EagI  (1641)
1 site
C G G C C G G C C G G C
NotI  (1641)
1 site
G C G G C C G C C G C C G G C G
SacII  (1641)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII
recognition sequence.
BglII  (1649)
1 site
A G A T C T T C T A G A
BamHI  (1655)
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  (1662)
1 site
A T C G A T T A G C T A
ClaI  (1662)
1 site
A T C G A T T A G C T A
SalI  (1667)
1 site
G T C G A C C A G C T G
AccI  (1668)
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  (1675)
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.
NdeI  (1680)
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  (1685)
1 site
T C T A G A A G A T C T
MfeI  (1784)
1 site
C A A T T G G T T A A C
HpaI  (1797)
1 site
G T T A A C C A A T T G
PsiI  (1817)
1 site
T T A T A A A A T A T T
AmpR
2820 .. 3680  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2820 .. 3611  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2820 .. 3680  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   3612 .. 3680  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2820 .. 3680  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
mCherry
335 .. 1045  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
mCherry
335 .. 1045  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
ori
2061 .. 2649  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2061 .. 2649  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES2
1053 .. 1639  =  587 bp
   Segment 1:  
   1053 .. 1627  =  575 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1053 .. 1639  =  587 bp
   Segment 2:  ATG  
   1628 .. 1630  =  3 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1053 .. 1639  =  587 bp
   Segment 3:  
   1631 .. 1639  =  9 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1053 .. 1639  =  587 bp
3 segments
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
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
3681 .. 3785  =  105 bp
AmpR promoter
3681 .. 3785  =  105 bp
SV40 poly(A) signal
1798 .. 1879  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1798 .. 1879  =  82 bp
SV40 polyadenylation signal
MCS
1640 .. 1690  =  51 bp
multiple cloning site
MCS
1640 .. 1690  =  51 bp
multiple cloning site
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
ATG
1628 .. 1630  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1628 .. 1630  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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