pTRE-Tight-BI-DsRed-Express

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

Sequence Author: Clontech (TaKaRa)

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HomePlasmidsMammalian Expression VectorspTRE-Tight-BI-DsRed-Express
EcoRI (3412) FspAI (3362) PstI - SbfI (3074) BbsI (2986) EcoO109I (2919) PflMI * (2895) CsiI - 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.
CsiI  (2888)
1 site		 A C C W G G T T G G W C C A
Sticky ends from different CsiI 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
Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
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
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
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
Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
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
2 segments
   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
2 segments
   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
ORF:  70 .. 354  =  285 bp
ORF:  94 amino acids  =  11.0 kDa
ORF:  1671 .. 1937  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  161 .. 418  =  258 bp
ORF:  85 amino acids  =  9.8 kDa
ORF:  1541 .. 2401  =  861 bp
ORF:  286 amino acids  =  31.5 kDa
ORF:  2729 .. 3490  =  762 bp
ORF:  253 amino acids  =  28.5 kDa
ORF:  2734 .. 3411  =  678 bp
ORF:  225 amino acids  =  25.7 kDa
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