pTet-On-Advanced

Vector for expressing the rtTA-Advanced transactivator in the Tet-On® Advanced system.

Sequence Author: Clontech (TaKaRa)

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BstZ17I (7089) AccI (7088) SfiI (6635) AvrII (6581) BmtI (6567) NheI (6563) BglII (6241) BclI * (6236) MscI (5999) RsrII (5562) BstBI (5397) Bsu36I (5202) BspEI * (5071) PflMI (4923) BsgI (4690) PspXI (4209) XmnI (3816) ScaI (3697) PvuI (3587) BsrGI (47) SpeI (103) SnaBI (444) Eco53kI (670) SacI (672) SacII (759) EcoRI (766) XbaI (778) PmlI (906) PspOMI (920) ApaI (924) FseI (1399) PpuMI (1815) BstEII (1911) HindIII (1998) Bpu10I (2021) PciI (2324) AhdI (3217) BsaI (3278) pTet-On-Advanced 7139 bp
BstZ17I  (7089)
1 site
G T A T A C C A T A T G
AccI  (7088)
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.
SfiI  (6635)
1 site
G G C C N N N N N G G C C C C G G N N N N N C C G G

Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
AvrII  (6581)
1 site
C C T A G G G G A T C C
BmtI  (6567)
1 site
G C T A G C C G A T C G
NheI  (6563)
1 site
G C T A G C C G A T C G
BglII  (6241)
1 site
A G A T C T T C T A G A
BclI  (6236)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
MscI  (5999)
1 site
T G G C C A A C C G G T
RsrII  (5562)
1 site
C G G W C C