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Plasmid Files

pTRE-Tight-BI-DsRed-Express

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

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pTRE-Tight-BI-DsRed-Express Sequence and MappTRE-Tight-BI-DsRed-Express.dna
Map and Sequence File   
Sequence Author:  Clontech
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 EcoRI (3412) FspAI (3362) PstI - SbfI (3074) BbsI (2986) EcoO109I (2919) PflMI * (2895) SexAI * (2888) MscI (2864) BsgI (2833) AleI (2783) BssHII (2762) XbaI (2728) SspI (2418) EarI (2409) XmnI (2213) ScaI (2094) TatI (2092) PvuI (1984) AseI (1786) PaeR7I - PspXI - XhoI (1) Acc65I (335) KpnI - TspMI - XmaI (339) SmaI (341) BamHI (344) MluI (362) NheI (368) BmtI (372) EagI - NotI (375) BspDI - ClaI (383) HindIII (388) SalI (394) AccI (395) EcoRV (402) PciI (721) NspI (725) DrdI (829) AlwNI (1137) BsaI (1675) BsrFI (1694) BglI (1734) pTRE-Tight-BI-DsRed-Express 3491 bp
EcoRI  (3412)
1 site
G A A T T C C T T A A G
FspAI  (3362)
1 site
R T G C G C A Y Y A C G C G T R
PstI  (3074)
1 site
C T G C A G G A C G T C
SbfI  (3074)
1 site
C C T G C A G G G G A C G T C C
BbsI  (2986)
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.
EcoO109I  (2919)
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.
PflMI  (2895)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
SexAI  (2888)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
MscI  (2864)
1 site
T G G C C A A C C G G T
BsgI  (2833)
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).
AleI  (2783)
1 site
C A C N N N N G T G G T G N N N N C A C
BssHII  (2762)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
XbaI  (2728)
1 site
T C T A G A A G A T C T
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
AseI  (1786)
1 site
A T T A A T T A A T T A
PaeR7I  (1)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1)
1 site
V C T C G A G B B G A G C T C V
XhoI  (1)
1 site
C T C G A G G A G C T C
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.
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.
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.
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.
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.
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.
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.
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
DsRed-Express
2734 .. 3411  =  678 bp
225 amino acids  =  25.7 kDa
Product: rapidly maturing tetrameric variant of
DsRed fluorescent protein
mammalian codon-optimized
DsRed-Express
2734 .. 3411  =  678 bp
225 amino acids  =  25.7 kDa
Product: rapidly maturing tetrameric variant of
DsRed fluorescent protein
mammalian codon-optimized
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
bidirectional TRE promoter
3422 .. 318  =  388 bp
Tet-responsive bidirectional promoter PTight-BI,
consisting of seven tet operator sequences flanked
on each side by the minimal CMV promoter
bidirectional TRE promoter
3422 .. 318  =  388 bp
Tet-responsive bidirectional promoter PTight-BI,
consisting of seven tet operator sequences flanked
on each side by the minimal CMV promoter
AmpR promoter
2402 .. 2506  =  105 bp
AmpR promoter
2402 .. 2506  =  105 bp
SV40 poly(A) signal
519 .. 600  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
519 .. 600  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2539 .. 2620  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2539 .. 2620  =  82 bp
SV40 polyadenylation signal
MCS
335 .. 405  =  71 bp
multiple cloning site
MCS
335 .. 405  =  71 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
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