Resources
Plasmid Files

pTRE-Cycle2

Vector for expressing mCherry while cycling the level of a second protein by alternating expression and rapid degradation.

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pTRE-Cycle2 Sequence and MappTRE-Cycle2.dna
Map and Sequence File   
Sequence Author:  Clontech
Download Free Trial Get SnapGene Viewer

 EcoRI (3813) AgeI (3807) NdeI (3802) SacII (3798) ApaI (3793) EcoO109I (3790) PspOMI (3789) BglII (3783) MscI (3745) BsaAI (3545) PstI - SbfI (3424) PflMI (3339) BbvCI - Bpu10I (3234) SgrAI - XcmI (3090) BsrGI (3071) XbaI (3060) AatII (2868) ZraI (2866) SspI (2750) EarI (2741) XmnI (2545) ScaI (2426) PvuI (2316) FspI (2168) AseI (2118) BsaI (2007) PaeR7I - PspXI - XhoI (1) Acc65I (335) KpnI (339) AarI - BfuAI - BspMI (351) BsmBI (377) BamHI (676) MluI (694) NheI (700) BmtI (704) EagI - NotI (707) BspDI - ClaI (715) HindIII (720) SalI (726) AccI (727) EcoRV (734) PciI (1053) pTRE-Cycle2 3892 bp
EcoRI  (3813)
1 site
G A A T T C C T T A A G
AgeI  (3807)
1 site
A C C G G T T G G C C A

AgeI quickly loses activity at 37°C, but can be used at 25°C for
long incubations.
NdeI  (3802)
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.
SacII  (3798)
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.
ApaI  (3793)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoO109I  (3790)
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  (3789)
1 site
G G G C C C C C C G G G
BglII  (3783)
1 site
A G A T C T T C T A G A
MscI  (3745)
1 site
T G G C C A A C C G G T
BsaAI  (3545)
1 site
Y A C G T R R T G C A Y
PstI  (3424)
1 site
C T G C A G G A C G T C
SbfI  (3424)
1 site
C C T G C A G G G G A C G T C C
PflMI  (3339)
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.
BbvCI  (3234)
1 site
C C T C A G C G G A G T C G
Bpu10I  (3234)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
SgrAI  (3090)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI
recognition sequence.
XcmI  (3090)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
BsrGI  (3071)
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.
XbaI  (3060)
1 site
T C T A G A A G A T C T
AatII  (2868)
1 site
G A C G T C C T G C A G
ZraI  (2866)
1 site
G A C G T C C T G C A G
SspI  (2750)
1 site
A A T A T T T T A T A A
EarI  (2741)
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  (2545)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2426)
1 site
A G T A C T T C A T G A
PvuI  (2316)
1 site
C G A T C G G C T A G C
FspI  (2168)
1 site
T G C G C A A C G C G T
AseI  (2118)
1 site
A T T A A T T A A T T A
BsaI  (2007)
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.
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
AarI  (351)
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  (351)
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  (351)
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.
BsmBI  (377)
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.
BamHI  (676)
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  (694)
1 site
A C G C G T T G C G C A
NheI  (700)
1 site
G C T A G C C G A T C G
BmtI  (704)
1 site
G C T A G C C G A T C G
EagI  (707)
1 site
C G G C C G G C C G G C
NotI  (707)
1 site
G C G G C C G C C G C C G G C G
BspDI  (715)
1 site
A T C G A T T A G C T A
ClaI  (715)
1 site
A T C G A T T A G C T A
HindIII  (720)
1 site
A A G C T T T T C G A A
SalI  (726)
1 site
G T C G A C C A G C T G
AccI  (727)
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  (734)
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  (1053)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
AmpR
1873 .. 2733  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 2:  
   1873 .. 2664  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1873 .. 2733  =  861 bp
286 amino acids  =  31.5 kDa
   Segment 1:  signal sequence  
   2665 .. 2733  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
1873 .. 2733  =  861 bp
286 amino acids  =  31.5 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
mCherry
3066 .. 3776  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
mCherry
3066 .. 3776  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
ori
1114 .. 1702  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
1114 .. 1702  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
bidirectional TRE promoter
3823 .. 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
3823 .. 318  =  388 bp
Tet-responsive bidirectional promoter PTight-BI,
consisting of seven tet operator sequences flanked
on each side by the minimal CMV promoter
DD
352 .. 675  =  324 bp
108 amino acids  =  11.9 kDa
   Segment 1:  
   352 .. 354  =  3 bp
   1 amino acid  =  149.2 Da
Product: destabilization domain that can be
stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
352 .. 675  =  324 bp
108 amino acids  =  11.9 kDa
   Segment 2:  
   355 .. 675  =  321 bp
   107 amino acids  =  11.8 kDa
Product: destabilization domain that can be
stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
DD
352 .. 675  =  324 bp
108 amino acids  =  11.9 kDa
2 segments
Product: destabilization domain that can be
stabilized by Shield1 in the ProteoTuner™ system
L106P mutant of FKBP12
AmpR promoter
2734 .. 2838  =  105 bp
AmpR promoter
2734 .. 2838  =  105 bp
SV40 poly(A) signal
851 .. 932  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
851 .. 932  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2871 .. 2952  =  82 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2871 .. 2952  =  82 bp
SV40 polyadenylation signal
MCS
676 .. 737  =  62 bp
multiple cloning site
MCS
676 .. 737  =  62 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
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter