pTRE-Tight-BI-AcGFP1

Vector for co-expressing AcGFP1 and another gene with the Tet-On® Advanced or Tet-Off® Advanced system.

Sequence Author: Clontech (TaKaRa)

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EcoRI (3454) PluTI (3440) SfoI (3438) NarI (3437) KasI (3436) MscI (3379) AleI (3272) BstEII (3265) Bpu10I (3257) Bsu36I (3181) PfoI * (3167) BssHII (3122) DraIII (3006) EcoNI (2827) StyI (2823) EcoO109I - PpuMI (2820) PflMI (2757) BtgI (2756) XbaI * (2728) AatII (2536) ZraI (2534) SspI (2418) EarI (2409) XmnI (2213) ScaI (2094) PvuI (1984) FspI (1836) AseI (1786) PaeR7I - PspXI - XhoI (1) Acc65I (335) KpnI - TspMI - XmaI (339) SmaI (341) BamHI (344) MluI (362) NheI (368) BmtI (372) EagI - NotI (375) BspDI - ClaI (383) HindIII (388) SalI (394) AccI (395) EcoRV (402) PciI (721) NspI (725) DrdI (829) BseYI (1025) PspFI (1029) AlwNI (1137) AhdI (1614) BmrI (1654) BsaI (1675) BglI (1734) pTRE-Tight-BI-AcGFP1 3533 bp
EcoRI  (3454)
1 site
G A A T T C C T T A A G
PluTI  (3440)
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.
SfoI  (3438)
1 site
G G C G C C C C G C G G
NarI  (3437)
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.
KasI  (3436)
1 site
G G C G C C C C G C G G
MscI  (3379)
1 site
T G G C C A A C C G G T
AleI  (3272)
1 site
C A C N N N N G T G G T G N N N N C A C
BstEII  (3265)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
Bpu10I  (3257)
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.
Bsu36I  (3181)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PfoI  (3167)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
BssHII  (3122)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
DraIII  (3006)
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.
EcoNI  (2827)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
StyI  (2823)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
EcoO109I  (2820)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
PpuMI  (2820)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
PflMI  (2757)
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.
BtgI  (2756)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
XbaI  (2728)
1 site
T C T A G A A G A T C T
* Blocked by Dam methylation.
AatII  (2536)
1 site
G A C G T C C T G C A G
ZraI  (2534)
1 site
G A C G T C C T G C A G
SspI  (2418)
1 site
A A T A T T T T A T A A
EarI  (2409)
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.
XmnI  (2213)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2094)
1 site
A G T A C T T C A T G A
PvuI  (1984)
1 site
C G A T C G G C T A G C
FspI  (1836)
1 site
T G C G C A A C G C G T
AseI  (1786)
1 site
A T T A A T T A A T T A
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
Acc65I  (335)
1 site
G G T A C C C C A T G G
KpnI  (339)
1 site
G G T A C C C C A T G G
TspMI  (339)
1 site
C C C G G G G G G C C C
XmaI  (339)
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  (341)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BamHI  (344)
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.
MluI  (362)
1 site
A C G C G T T G C G C A
NheI  (368)
1 site
G C T A G C C G A T C G
BmtI  (372)
1 site
G C T A G C C G A T C G
EagI  (375)
1 site
C G G C C G G C C G G C
NotI  (375)
1 site
G C G G C C G C C G C C G G C G
BspDI  (383)
1 site
A T C G A T T A G C T A
ClaI  (383)
1 site
A T C G A T T A G C T A
HindIII  (388)
1 site
A A G C T T T T C G A A
SalI  (394)
1 site
G T C G A C C A G C T G
AccI  (395)
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.
EcoRV  (402)
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.
PciI  (721)
1 site
A C A T G T T