pTRE3G-mCherry

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

Sequence Author: Clontech (TaKaRa)

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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) 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) BfuAI - BspMI - PaqCI (1447) DraIII (1471) Acc65I (1549) KpnI (1553) BmgBI (1651) EcoRV (1703) MluI (1707) NdeI (1714) NheI (1719) BmtI (1723) BamHI (1731) BtgZI (1768) BsaBI * (2405) 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
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

Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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
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.
PaqCI  (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 PaqCI recognition sequence.
Sticky ends from different PaqCI sites may not be compatible.
Cleavage can be improved with PaqCI Activator.
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