pTRE-Cycle3

Vector for expressing ZsGreen1 while cycling the level of a second protein by alternating expression and rapid degradation.

Sequence Author: Clontech (TaKaRa)

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EcoRI (3793) BclI * (3699) SgrAI (3694) PflFI - Tth111I (3550) NaeI (3519) NgoMIV (3517) PluTI (3482) SfoI (3480) NarI (3479) KasI (3478) BseRI (3427) BmgBI (3294) BsrGI (3231) BtgZI (3128) ApaI (3089) EcoO109I (3086) PspOMI (3085) BglII (3079) PstI (3070) XbaI (3060) AatII (2868) ZraI (2866) SspI (2750) EarI (2741) XmnI (2545) ScaI (2426) PvuI (2316) FspI (2168) AseI (2118) BsaI (2007) PaeR7I - PspXI - XhoI (1) Acc65I (335) KpnI (339) 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) EcoRV (734) PciI (1053) DrdI (1161) AlwNI (1469) pTRE-Cycle3 3872 bp
EcoRI  (3793)
1 site
G A A T T C C T T A A G
BclI  (3699)
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.
SgrAI  (3694)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
PflFI  (3550)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3550)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
NaeI  (3519)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (3517)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
PluTI  (3482)
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  (3480)
1 site
G G C G C C C C G C G G
NarI  (3479)
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  (3478)
1 site
G G C G C C C C G C G G
BseRI  (3427)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BmgBI  (3294)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
BsrGI  (3231)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
BtgZI  (3128)
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.
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
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
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
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.
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
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