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Plasmid Files

pTRE-Dual1

Vector for co-expressing two genes with the Tet-On® Advanced or Tet-Off® Advanced system.

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pTRE-Dual1 Sequence and MappTRE-Dual1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 ZraI (3128) SspI (3012) EarI (3003) ScaI (2688) TatI (2686) PvuI (2578) FspI (2430) AseI (2380) NmeAIII (2356) BsaI (2269) AhdI (2208) AlwNI (1731) PspFI (1623) AatII (3130) StuI (277) Eco53kI (292) SacI (294) EcoRI (323) PstI (333) TspMI - XmaI (335) SmaI (337) PacI (345) PvuII (351) MluI (355) NheI (361) BmtI (365) PspOMI (481) ApaI (485) AvrII - StyI (519) HindIII (592) BsaAI - PmlI (684) AarI - BfuAI - BspMI (707) DraIII (731) BseRI (758) Acc65I (809) KpnI (813) PflMI (821) BmgBI (911) MscI (946) EagI - NotI - SacII (956) BglII (964) BamHI (970) BspDI - ClaI (977) SalI (982) AccI (983) EcoRV (990) NdeI (995) XbaI (1000) MfeI (1099) HpaI (1112) PsiI (1132) DrdI (1423) BseYI (1619) pTRE-Dual1 3199 bp
ZraI  (3128)
1 site
G A C G T C C T G C A G
SspI  (3012)
1 site
A A T A T T T T A T A A
EarI  (3003)
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  (2688)
1 site
A G T A C T T C A T G A
TatI  (2686)
1 site
W G T A C W W C A T G W
PvuI  (2578)
1 site
C G A T C G G C T A G C
FspI  (2430)
1 site
T G C G C A A C G C G T
AseI  (2380)
1 site
A T T A A T T A A T T A
NmeAIII  (2356)
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).
BsaI  (2269)
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.
AhdI  (2208)
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.
AlwNI  (1731)
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.
PspFI  (1623)
1 site
C C C A G C G G G T C G
AatII  (3130)
1 site
G A C G T C C T G C A G
StuI  (277)
1 site
A G G C C T T C C G G A
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  (333)
1 site
C T G C A G G A C G T C
TspMI  (335)
1 site
C C C G G G G G G C C C
XmaI  (335)
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  (337)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PacI  (345)
1 site
T T A A T T A A A A T T A A T T
PvuII  (351)
1 site
C A G C T G G T C G A C
MluI  (355)
1 site
A C G C G T T G C G C A
NheI  (361)
1 site
G C T A G C C G A T C G
BmtI  (365)
1 site
G C T A G C C G A T C G
PspOMI  (481)
1 site
G G G C C C C C C G G G
ApaI  (485)
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  (519)
1 site
C C T A G G G G A T C C
StyI  (519)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
HindIII  (592)
1 site
A A G C T T T T C G A A
BsaAI  (684)
1 site
Y A C G T R R T G C A Y
PmlI  (684)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (707)
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  (707)
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  (707)
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  (731)
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.
BseRI  (758)
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.
Acc65I  (809)
1 site
G G T A C C C C A T G G
KpnI  (813)
1 site
G G T A C C C C A T G G
PflMI  (821)
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.
BmgBI  (911)
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.
MscI  (946)
1 site
T G G C C A A C C G G T
EagI  (956)
1 site
C G G C C G G C C G G C
NotI  (956)
1 site
G C G G C C G C C G C C G G C G
SacII  (956)
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  (964)
1 site
A G A T C T T C T A G A
BamHI  (970)
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  (977)
1 site
A T C G A T T A G C T A
ClaI  (977)
1 site
A T C G A T T A G C T A
SalI  (982)
1 site
G T C G A C C A G C T G
AccI  (983)
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  (990)
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  (995)
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  (1000)
1 site
T C T A G A A G A T C T
MfeI  (1099)
1 site
C A A T T G G T T A A C
HpaI  (1112)
1 site
G T T A A C C A A T T G
PsiI  (1132)
1 site
T T A T A A A A T A T T
DrdI  (1423)
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  (1619)
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.
AmpR
2135 .. 2995  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   2135 .. 2926  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2135 .. 2995  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   2927 .. 2995  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2135 .. 2995  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1376 .. 1964  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1376 .. 1964  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES2
368 .. 954  =  587 bp
   Segment 1:  
   368 .. 942  =  575 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
368 .. 954  =  587 bp
   Segment 2:  ATG  
   943 .. 945  =  3 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
368 .. 954  =  587 bp
   Segment 3:  
   946 .. 954  =  9 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
368 .. 954  =  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
2996 .. 3100  =  105 bp
AmpR promoter
2996 .. 3100  =  105 bp
SV40 poly(A) signal
1113 .. 1194  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1113 .. 1194  =  82 bp
SV40 polyadenylation signal
MCS 2
955 .. 1005  =  51 bp
multiple cloning site 2
MCS 2
955 .. 1005  =  51 bp
multiple cloning site 2
MCS 1
323 .. 366  =  44 bp
multiple cloning site 1
MCS 1
323 .. 366  =  44 bp
multiple cloning site 1
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
943 .. 945  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
943 .. 945  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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