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Plasmid Files

pTRE3G-BI-ZsGreen1

Vector for doxycycline-inducible expression of a gene together with the ZsGreen1 fluorescent protein.

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pTRE3G-BI-ZsGreen1 Sequence and MappTRE3G-BI-ZsGreen1.dna
Map and Sequence File   
Sequence Author:  Clontech
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 NcoI (3538) BclI * (3454) PflFI - Tth111I (3305) BseRI (3182) BmgBI (3049) BfuAI - BspMI (3025) BsrGI (2986) BtgZI (2883) PstI (2844) PshAI (2842) XbaI (2834) AatII (2642) ZraI (2640) SspI (2524) XmnI (2319) ScaI (2200) PvuI (2090) FspI (1942) AseI (1892) EcoRI (3545) StuI (40) PaeR7I - PspXI - XhoI (70) HindIII (322) EcoO109I - PspOMI (462) ApaI (466) BglII (468) BamHI (480) BspDI * - ClaI * (487) EagI - NotI (495) MreI (500) EcoRV (508) AflIII - PciI (827) DrdI (935) BseYI (1131) PspFI (1135) AlwNI (1243) BsaI (1781) pTRE3G-BI-ZsGreen1 3555 bp
NcoI  (3538)
1 site
C C A T G G G G T A C C
BclI  (3454)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
PflFI  (3305)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3305)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BseRI  (3182)
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.
BmgBI  (3049)
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.
BfuAI  (3025)
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  (3025)
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.
BsrGI  (2986)
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.
BtgZI  (2883)
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.
PstI  (2844)
1 site
C T G C A G G A C G T C
PshAI  (2842)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
XbaI  (2834)
1 site
T C T A G A A G A T C T
AatII  (2642)
1 site
G A C G T C C T G C A G
ZraI  (2640)
1 site
G A C G T C C T G C A G
SspI  (2524)
1 site
A A T A T T T T A T A A
XmnI  (2319)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2200)
1 site
A G T A C T T C A T G A
PvuI  (2090)
1 site
C G A T C G G C T A G C
FspI  (1942)
1 site
T G C G C A A C G C G T
AseI  (1892)
1 site
A T T A A T T A A T T A
EcoRI  (3545)
1 site
G A A T T C C T T A A G
StuI  (40)
1 site
A G G C C T T C C G G A
PaeR7I  (70)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (70)
1 site
V C T C G A G B B G A G C T C V
XhoI  (70)
1 site
C T C G A G G A G C T C
HindIII  (322)
1 site
A A G C T T T T C G A A
EcoO109I  (462)
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.
PspOMI  (462)
1 site
G G G C C C C C C G G G
ApaI  (466)
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  (468)
1 site
A G A T C T T C T A G A
BamHI  (480)
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  (487)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (487)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EagI  (495)
1 site
C G G C C G G C C G G C
NotI  (495)
1 site
G C G G C C G C C G C C G G C G
MreI  (500)
1 site
C G C C G G C G G C G G C C G C
EcoRV  (508)
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.
AflIII  (827)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (827)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (935)
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  (1131)
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.
PspFI  (1135)
1 site
C C C A G C G G G T C G
AlwNI  (1243)
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  (1781)
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.
AmpR
1647 .. 2507  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   1647 .. 2438  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1647 .. 2507  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   2439 .. 2507  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1647 .. 2507  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ZsGreen1
2846 .. 3541  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ZsGreen1
2846 .. 3541  =  696 bp
231 amino acids  =  26.1 kDa
Product: Zoanthus green fluorescent protein
mammalian codon-optimized
ori
888 .. 1476  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
888 .. 1476  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
TRE3G BI promoter
1 .. 451  =  451 bp
3rd-generation Tet-responsive bidirectional
promoter that can be activated by binding of Tet-On
3G
TRE3G BI promoter
1 .. 451  =  451 bp
3rd-generation Tet-responsive bidirectional
promoter that can be activated by binding of Tet-On
3G
AmpR promoter
2508 .. 2612  =  105 bp
AmpR promoter
2508 .. 2612  =  105 bp
SV40 poly(A) signal
625 .. 706  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
625 .. 706  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2645 .. 2726  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2645 .. 2726  =  82 bp
SV40 polyadenylation signal
MCS
452 .. 511  =  60 bp
multiple cloning site
MCS
452 .. 511  =  60 bp
multiple cloning site
tet operator
81 .. 99  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
81 .. 99  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
117 .. 135  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
117 .. 135  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
153 .. 171  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
153 .. 171  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
189 .. 207  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
189 .. 207  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
225 .. 243  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
225 .. 243  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
261 .. 279  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
261 .. 279  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
297 .. 315  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
297 .. 315  =  19 bp
bacterial operator O2 for the tetR and tetA genes
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