pTRE-Dual1

Vector for co-expressing two genes with the Tet-On® Advanced or Tet-Off® Advanced system.

Sequence Author: Clontech (TaKaRa)

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ZraI (3128) SspI (3012) EarI (3003) ScaI (2688) TatI (2686) PvuI (2578) FspI (2430) AseI (2380) NmeAIII (2356) BsaI (2269) AhdI (2208) AlwNI (1731) PspFI (1623) AatII (3130) StuI (277) Eco53kI (292) SacI (294) EcoRI (323) PstI (333) TspMI - XmaI (335) SmaI (337) PacI (345) PvuII (351) MluI (355) NheI (361) BmtI (365) PspOMI (481) ApaI (485) AvrII - StyI (519) HindIII (592) BsaAI - PmlI (684) BfuAI - BspMI - PaqCI (707) DraIII (731) BseRI (758) Acc65I (809) KpnI (813) PflMI (821) BmgBI (911) MscI (946) EagI - NotI - SacII (956) BglII (964) BamHI (970) BspDI - ClaI (977) SalI (982) AccI (983) EcoRV (990) NdeI (995) XbaI (1000) MfeI (1099) HpaI (1112) PsiI (1132) DrdI (1423) BseYI (1619) pTRE-Dual1 3199 bp
ZraI  (3128)
1 site
G A C G T C C T G C A G
SspI  (3012)
1 site
A A T A T T T T A T A A
EarI  (3003)
1 site
C T C T T C N G A G A A G N N N N

Cleavage may be enhanced when more than one copy of the EarI recognition sequence is present.
Sticky ends from different EarI sites may not be compatible.
ScaI  (2688)
1 site
A G T A C T T C A T G A
TatI  (2686)
1 site
W G T A C W W C A T G W
PvuI  (2578)
1 site
C G A T C G G C T A G C
FspI  (2430)
1 site
T G C G C A A C G C G T
AseI  (2380)
1 site
A T T A A T T A A T T A
NmeAIII  (2356)
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  (2269)
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  (2208)
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  (1731)
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  (1623)
1 site
C C C A G C G G G T C G
AatII  (3130)
1 site
G A C G T C C T G C A G
StuI  (277)
1 site
A G G C C T T C C G G A
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
EcoRI  (323)
1 site
G A A T T C C T T A A G
PstI  (333)
1 site
C T G C A G G A C G T C
TspMI  (335)
1 site
C C C G G G G G G C C C
XmaI  (335)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (337)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
PacI  (345)
1 site
T T A A T T A A A A T T A A T T
PvuII  (351)
1 site
C A G C T G G T C G A C
MluI  (355)
1 site
A C G C G T T G C G C A
NheI  (361)
1 site
G C T A G C C G A T C G
BmtI  (365)
1 site
G C T A G C C G A T C G
PspOMI  (481)
1 site
G G G C C C C C C G G G
ApaI  (485)
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  (519)
1 site
C C T A G G G G A T C C
StyI  (519)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
HindIII  (592)
1 site
A A G C T T T T C G A A
BsaAI  (684)
1 site
Y A C G T R R T G C A Y
PmlI  (684)
1 site
C A C G T G G T G C A C
BfuAI  (707)
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  (707)
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  (707)
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  (731)
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.
BseRI  (758)
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.
Acc65I  (809)
1 site
G G T A C C C C A T G G
KpnI  (813)
1 site
G G T A C C C C A T G G
PflMI  (821)
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.
BmgBI  (911)
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.
MscI  (946)
1 site
T G G C C A A C C G G T
EagI  (956)
1 site
C G G C C G G C C G G C
NotI  (956)
1 site
G C G G C C G C C G C C G G C