pTRE-Cycle1

Bidirectional expression vector for cycling the level of a protein by alternating expression and rapid degradation.

Sequence Author: Clontech (TaKaRa)

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EcoRI (3109) AgeI (3103) NdeI (3098) SacII (3094) ApaI (3089) EcoO109I (3086) PspOMI (3085) BglII (3079) Bsu36I (3073) PstI (3070) XbaI (3060) AatII (2868) ZraI (2866) SspI (2750) EarI (2741) XmnI (2545) ScaI (2426) TatI (2424) PvuI (2316) FspI (2168) AseI (2118) NmeAIII (2094) BsaI (2007) AhdI (1946) PaeR7I - PspXI - XhoI (1) Acc65I (335) KpnI (339) NcoI - StyI (350) BfuAI - BspMI - PaqCI (351) BsmBI - Esp3I (377) BsgI (379) BbsI (639) BamHI (676) PvuII (690) MluI (694) NheI (700) BmtI (704) EagI - NotI (707) BspDI - ClaI (715) HindIII (720) SalI (726) AccI (727) EcoRV (734) PciI (1053) DrdI (1161) HaeII (1301) AlwNI (1469) pTRE-Cycle1 3188 bp
EcoRI  (3109)
1 site
G A A T T C C T T A A G
AgeI  (3103)
1 site
A C C G G T T G G C C A
NdeI  (3098)
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.
SacII  (3094)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
ApaI  (3089)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
EcoO109I  (3086)
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.
PspOMI  (3085)
1 site
G G G C C C C C C G G G
BglII  (3079)
1 site
A G A T C T T C T A G A
Bsu36I  (3073)
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.
PstI  (3070)
1 site
C T G C A G G A C G T C
XbaI  (3060)
1 site
T C T A G A A G A T C T
AatII  (2868)
1 site
G A C G T C C T G C A G
ZraI  (2866)
1 site
G A C G T C C T G C A G
SspI  (2750)
1 site
A A T A T T T T A T A A
EarI  (2741)
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  (2545)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (2426)
1 site
A G T A C T T C A T G A
TatI  (2424)
1 site
W G T A C W W C A T G W
PvuI  (2316)
1 site
C G A T C G G C T A G C
FspI  (2168)
1 site
T G C G C A A C G C G T
AseI  (2118)
1 site
A T T A A T T A A T T A
NmeAIII  (2094)
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  (2007)
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  (1946)
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.
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
NcoI  (350)
1 site
C C A T G G G G T A C C
StyI  (350)
1 site
C C W W G G G G W W C C

Sticky ends from different StyI sites may not be compatible.
BfuAI  (351)
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  (351)
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  (351)
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.
BsmBI  (377)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (377)
1 site
C G T C T C N G C A G A G N ( N ) 4

Sticky ends from different Esp3I sites may not be compatible.
BsgI  (379)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BbsI  (639)
1 site
G A A G A C N N C T T C T G N N ( N ) 4

Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BamHI  (676)
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.
PvuII  (690)
1 site
C A G C T G G T C G A C
MluI  (694)
1 site
A C G C G T T G C G C A
NheI  (700)
1 site