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Plasmid Files

pTRE3G-mCherry

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

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pTRE3G-mCherry Sequence and MappTRE3G-mCherry.dna
Map and Sequence File   
Sequence Author:  Clontech
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 EcoRI (4710) AatII (4639) ZraI (4637) ScaI (4197) PvuI (4087) FspI (3939) BsaI (3778) BspQI - SapI (2708) HpaI (2506) MfeI (2493) BsaBI * (2405) PaeR7I - PspXI - XhoI (1) Eco53kI (292) SacI (294) SalI (383) AccI (384) NcoI (822) BbvCI - Bpu10I (927) PvuII (967) AleI (1031) SgrAI (1070) XcmI (1075) BsrGI (1089) EagI (1100) NgoMIV (1102) NaeI (1104) FseI (1106) PspOMI (1221) ApaI (1225) AvrII (1259) PmlI (1424) AarI - BfuAI - BspMI (1447) DraIII (1471) Acc65I (1549) KpnI (1553) BmgBI (1651) EcoRV (1703) MluI (1707) NdeI (1714) NheI (1719) BmtI (1723) BamHI (1731) BtgZI (1768) pTRE3G-mCherry 4715 bp
EcoRI  (4710)
1 site
G A A T T C C T T A A G
AatII  (4639)
1 site
G A C G T C C T G C A G
ZraI  (4637)
1 site
G A C G T C C T G C A G
ScaI  (4197)
1 site
A G T A C T T C A T G A
PvuI  (4087)
1 site
C G A T C G G C T A G C
FspI  (3939)
1 site
T G C G C A A C G C G T
BsaI  (3778)
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.
BspQI  (2708)
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  (2708)
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.
HpaI  (2506)
1 site
G T T A A C C A A T T G
MfeI  (2493)
1 site
C A A T T G G T T A A C
BsaBI  (2405)
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.
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
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.
NcoI  (822)
1 site
C C A T G G G G T A C C
BbvCI  (927)
1 site
C C T C A G C G G A G T C G
Bpu10I  (927)
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.
PvuII  (967)
1 site
C A G C T G G T C G A C
AleI  (1031)
1 site
C A C N N N N G T G G T G N N N N C A C
SgrAI  (1070)
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  (1075)
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  (1089)
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.
EagI  (1100)
1 site
C G G C C G G C C G G C
NgoMIV  (1102)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV
recognition sequence.
NaeI  (1104)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI
recognition sequence.
FseI  (1106)
1 site
G G C C G G C C C C G G C C G G

FseI gradually loses activity when stored at -20°C.
PspOMI  (1221)
1 site
G G G C C C C C C G G G
ApaI  (1225)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
AvrII  (1259)
1 site
C C T A G G G G A T C C
PmlI  (1424)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
AarI  (1447)
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  (1447)
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  (1447)
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.
DraIII  (1471)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
Acc65I  (1549)
1 site
G G T A C C C C A T G G
KpnI  (1553)
1 site
G G T A C C C C A T G G
BmgBI  (1651)
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.
EcoRV  (1703)
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.
MluI  (1707)
1 site
A C G C G T T G C G C A
NdeI  (1714)
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  (1719)
1 site
G C T A G C C G A T C G
BmtI  (1723)
1 site
G C T A G C C G A T C G
BamHI  (1731)
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  (1768)
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.
AmpR
3644 .. 4504  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   3644 .. 4435  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3644 .. 4504  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   4436 .. 4504  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
3644 .. 4504  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
mCherry
389 .. 1099  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
mCherry
389 .. 1099  =  711 bp
236 amino acids  =  26.7 kDa
Product: monomeric derivative of DsRed fluorescent
protein
mammalian codon-optimized
ori
2885 .. 3473  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
2885 .. 3473  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
IRES2
1107 .. 1694  =  588 bp
   Segment 1:  
   1107 .. 1682  =  576 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1107 .. 1694  =  588 bp
   Segment 2:  ATG  
   1683 .. 1685  =  3 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1107 .. 1694  =  588 bp
   Segment 3:  
   1686 .. 1694  =  9 bp
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
IRES2
1107 .. 1694  =  588 bp
3 segments
internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)
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
SV40 poly(A) signal
2507 .. 2628  =  122 bp
SV40 polyadenylation signal
SV40 poly(A) signal
2507 .. 2628  =  122 bp
SV40 polyadenylation signal
AmpR promoter
4505 .. 4609  =  105 bp
AmpR promoter
4505 .. 4609  =  105 bp
small t intron
1867 .. 1932  =  66 bp
simian virus 40 (SV40) small t antigen intron
small t intron
1867 .. 1932  =  66 bp
simian virus 40 (SV40) small t antigen intron
MCS
1701 .. 1736  =  36 bp
multiple cloning site
MCS
1701 .. 1736  =  36 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
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
ATG
1683 .. 1685  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
ATG
1683 .. 1685  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon for translation from IRES2
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