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Plasmid Files

pTRE3G-Hom1

Mammalian vector encoding a doxycycline-inducible FKBP homodimerizer domain, for creating soluble fusion proteins that can be dimerized with a drug. 

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pTRE3G-Hom1 Sequence and MappTRE3G-Hom1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 EcoRI (3770) AatII (3699) ZraI (3697) XmnI (3376) ScaI (3257) TatI (3255) FspI (2999) NmeAIII (2925) BglI (2897) BsrFI (2857) BsaI (2838) AhdI (2777) AlwNI (2300) DrdI (1992) PaeR7I - PspXI - XhoI (1) HindIII (253) KasI (274) NarI (275) SfoI (276) PluTI (278) Eco53kI (292) SacI (294) SalI (383) AccI (384) PspOMI (393) ApaI (397) BglII (399) BspDI * - ClaI * (406) EagI (411) EcoRV (419) ATG AarI (431) BsmBI (457) TspMI - XmaI (550) SmaI (552) MluI (756) NdeI (763) NheI (768) BmtI (772) PstI - SbfI (778) BamHI (780) BtgZI (828) PflMI (1002) StyI (1150) BsaBI * (1465) MfeI (1553) HpaI (1566) BspQI - SapI (1768) PciI (1884) pTRE3G-Hom1 3775 bp
EcoRI  (3770)
1 site
G A A T T C C T T A A G
AatII  (3699)
1 site
G A C G T C C T G C A G
ZraI  (3697)
1 site
G A C G T C C T G C A G
XmnI  (3376)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (3257)
1 site
A G T A C T T C A T G A
TatI  (3255)
1 site
W G T A C W W C A T G W
FspI  (2999)
1 site
T G C G C A A C G C G T
NmeAIII  (2925)
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).
BglI  (2897)
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.
BsrFI  (2857)
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.
BsaI  (2838)
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  (2777)
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  (2300)
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.
DrdI  (1992)
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.
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
HindIII  (253)
1 site
A A G C T T T T C G A A
KasI  (274)
1 site
G G C G C C C C G C G G
NarI  (275)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI
recognition sequence.
SfoI  (276)
1 site
G G C G C C C C G C G G
PluTI  (278)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI
recognition sequence.
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
SalI  (383)
1 site
G T C G A C C A G C T G
AccI  (384)
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.
PspOMI  (393)
1 site
G G G C C C C C C G G G
ApaI  (397)
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  (399)
1 site
A G A T C T T C T A G A
BspDI  (406)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (406)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EagI  (411)
1 site
C G G C C G G C C G G C
EcoRV  (419)
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.
AarI  (431)
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.
BsmBI  (457)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
TspMI  (550)
1 site
C C C G G G G G G C C C
XmaI  (550)
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  (552)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
MluI  (756)
1 site
A C G C G T T G C G C A
NdeI  (763)
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.
NheI  (768)
1 site
G C T A G C C G A T C G
BmtI  (772)
1 site
G C T A G C C G A T C G
PstI  (778)
1 site
C T G C A G G A C G T C
SbfI  (778)
1 site
C C T G C A G G G G A C G T C C
BamHI  (780)
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.
BtgZI  (828)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its
electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
PflMI  (1002)
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.
StyI  (1150)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BsaBI  (1465)
1 site
G A T N N N N A T C C T A N N N N T A G
* Blocked by Dam methylation.
MfeI  (1553)
1 site
C A A T T G G T T A A C
HpaI  (1566)
1 site
G T T A A C C A A T T G
BspQI  (1768)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (1768)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be
mixed before removing an aliquot.
PciI  (1884)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
2704 .. 3564  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   2704 .. 3495  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2704 .. 3564  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   3496 .. 3564  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
2704 .. 3564  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
1945 .. 2533  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1945 .. 2533  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
TRE3G promoter
4 .. 382  =  379 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G
TRE3G promoter
4 .. 382  =  379 bp
3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On® 3G
DmrB
435 .. 755  =  321 bp
107 amino acids  =  11.8 kDa
Product: F36V mutant of FK506-binding protein
FKBP12
binds synthetic ligands that do not bind wild-type
FKBP
DmrB
435 .. 755  =  321 bp
107 amino acids  =  11.8 kDa
Product: F36V mutant of FK506-binding protein
FKBP12
binds synthetic ligands that do not bind wild-type
FKBP
SV40 poly(A) signal
1567 .. 1688  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
1567 .. 1688  =  122 bp
SV40 polyadenylation signal
AmpR promoter
3565 .. 3669  =  105 bp
AmpR promoter
3565 .. 3669  =  105 bp
small t intron
927 .. 992  =  66 bp
simian virus 40 (SV40) small t antigen intron
small t intron
927 .. 992  =  66 bp
simian virus 40 (SV40) small t antigen intron
5' MCS
383 .. 422  =  40 bp
multiple cloning site upstream of DmrB
5' MCS
383 .. 422  =  40 bp
multiple cloning site upstream of DmrB
MCS
756 .. 785  =  30 bp
multiple cloning site
MCS
756 .. 785  =  30 bp
multiple cloning site
ATG
423 .. 425  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
423 .. 425  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
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
84 .. 102  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
84 .. 102  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
120 .. 138  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
120 .. 138  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
156 .. 174  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
156 .. 174  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
192 .. 210  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
192 .. 210  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
228 .. 246  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
228 .. 246  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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