pTet-DualOFF

Vector for co-expressing ZsGreen1 with the Tet-Off® Advanced transcriptional activator.

Sequence Author: Clontech (TaKaRa)

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SspI (4719) EarI (4710) ScaI (4395) PvuI (4285) FspI (4137) AseI (4087) BsaI (3976) AlwNI (3438) DrdI (3130) PsiI (2839) HpaI (2819) BsaBI * (2718) EcoRV (2703) BspDI - ClaI (2684) NotI (2676) Bpu10I (2671) BsrGI (2529) PaeR7I - PspXI - XhoI (1) SnaBI (289) Eco53kI (515) SacI (517) BsmBI - Esp3I (538) SacII (604) BamHI (607) PvuII (621) MluI (625) StuI (704) NruI (914) FseI (1258) TspMI - XmaI (1372) SmaI (1374) EcoRI (1384) AvrII (1543) PaqCI (1731) DraIII (1755) Acc65I (1833) KpnI (1837) PflMI (1845) MscI (1970) BstXI (1978) BclI * (2061) SgrAI (2066) PflFI - Tth111I (2213) KasI (2282) NarI (2283) SfoI (2284) PluTI (2286) pTet-DualOFF 4906 bp
SspI  (4719)
1 site
A A T A T T T T A T A A
EarI  (4710)
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  (4395)
1 site
A G T A C T T C A T G A
PvuI  (4285)
1 site
C G A T C G G C T A G C
FspI  (4137)
1 site
T G C G C A A C G C G T
AseI  (4087)
1 site
A T T A A T T A A T T A
BsaI  (3976)
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.
AlwNI  (3438)
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.
DrdI  (3130)
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.
PsiI  (2839)
1 site
T T A T A A A A T A T T
HpaI  (2819)
1 site
G T T A A C C A A T T G
BsaBI  (2718)
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.
EcoRV  (2703)
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.
BspDI  (2684)
1 site
A T C G A T T A G C T A
ClaI  (2684)
1 site
A T C G A T T A G C T A
NotI  (2676)
1 site
G C G G C C G C C G C C G G C G
Bpu10I  (2671)
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.
BsrGI  (2529)
1 site
T G T A C A A C A T G T