pTRE3G-IRES

Vector for doxycycline-inducible expression of two genes in cells containing the Tet-On® 3G transactivator protein.

Sequence Author: Clontech (TaKaRa)

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EcoRI (4021) AatII (3950) ZraI (3948) ScaI (3508) TatI (3506) FspI (3250) NmeAIII (3176) BsaI (3089) AhdI (3028) AlwNI (2551) PspFI (2443) BseYI (2439) DrdI (2243) BspQI - SapI (2019) AvaI - BsoBI - PaeR7I - PspXI - XhoI (1) KasI (274) NarI (275) SfoI (276) PluTI (278) Eco53kI (292) SacI (294) SalI (383) AccI (384) BglII (399) BspDI * - ClaI * (406) EagI (411) NgoMIV (413) NaeI (415) FseI (417) AvrII (570) BsaAI - PmlI (735) BfuAI - BspMI - PaqCI (758) DraIII (782) Acc65I (860) KpnI (864) BtgI (953) BmgBI (962) MscI (997) EcoRV (1014) MluI (1018) NdeI (1025) NheI (1030) BmtI (1034) PstI - SbfI (1040) BamHI (1042) BtgZI (1079) BsgI (1485) BsaBI * (1716) MfeI (1804) HpaI (1817) pTRE3G-IRES 4026 bp
EcoRI  (4021)
1 site
G A A T T C C T T A A G
AatII  (3950)
1 site
G A C G T C C T G C A G
ZraI  (3948)
1 site
G A C G T C C T G C A G
ScaI  (3508)
1 site
A G T A C T T C A T G A
TatI  (3506)
1 site
W G T A C W W C A T G W
FspI  (3250)
1 site
T G C G C A A C G C G T
NmeAIII  (3176)
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).
BsaI  (3089)
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  (3028)
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  (2551)
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.
PspFI  (2443)
1 site
C C C A G C G G G T C G
BseYI  (2439)
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.
DrdI  (2243)
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.
BspQI  (2019)
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  (2019)
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.
AvaI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (1)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°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
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.
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
NgoMIV  (413)
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  (415)
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  (417)
1 site
G G C C G G C C C C