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pTRE-Tight-BI-ZsGreen1

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

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pTRE-Tight-BI-ZsGreen1.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Clontech (TaKaRa)
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EcoRI (3430) BclI * (3342) NdeI (3111) Bpu10I (2959) EcoNI (2917) BtgZI (2781) BspEI * (2749) XbaI * (2728) AatII (2536) ZraI (2534) SspI (2418) EarI (2409) XmnI (2213) ScaI (2094) TatI (2092) PvuI (1984) FspI (1836) NmeAIII (1762) 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) PciI (721) DrdI (829) HaeII (969) BseYI (1025) PspFI (1029) AlwNI (1137) BsaI (1675) BsrFI (1694) pTRE-Tight-BI-ZsGreen1 3509 bp
EcoRI  (3430)
1 site
G A A T T C C T T A A G
BclI  (3342)
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.
NdeI  (3111)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
Bpu10I  (2959)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I recognition sequence.
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.
EcoNI  (2917)
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.
BtgZI  (2781)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
BspEI  (2749)
1 site
T C C G G A A G G C C T
* Blocked by Dam methylation.
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

Efficient cleavage requires at least two copies of the EarI recognition sequence.
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
TatI  (2092)
1 site
W G T A C W W C A T G W
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
NmeAIII  (1762)
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).
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

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
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