pd2ECFP-N1

Vector for fusing destabilized ECFP, with an in vivo half-life of ~2 hours, to the C-terminus of a partner protein.

Sequence Author: Clontech (TaKaRa)

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AflIII - PciI (4804) ApaLI (4490) EcoO109I (3984) BsaI (3875) RsrII (3402) BsrDI (3119) PflFI - Tth111I (3004) FspI (2988) MscI (2968) PluTI (2889) SfoI (2887) NarI (2886) KasI (2885) BspDI * - ClaI * (2726) StuI (2707) SfiI (2661) CsiI - SexAI * (2475) AseI (7) NdeI (234) SnaBI (340) NheI (591) BmtI (595) AfeI (596) BglII (609) PaeR7I - XhoI (613) Eco53kI (618) SacI (620) EcoRI (629) SalI (639) AccI (640) Acc65I (645) KpnI (649) SacII (652) PspOMI (653) TspMI - XmaI (656) ApaI (657) SmaI (658) BamHI (660) AgeI (666) BsrGI (1388) BlpI (1399) NotI (1530) XbaI * (1540) MfeI (1636) HpaI (1649) AflII (1768) DraIII (2002) pd2ECFP-N1 4862 bp
AflIII  (4804)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (4804)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
ApaLI  (4490)
1 site
G T G C A C C A C G T G
EcoO109I  (3984)
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.
BsaI  (3875)
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.
RsrII  (3402)
1 site
C G G W C C G G C C W G G C

Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BsrDI  (3119)
1 site
G C A A T G N N C G T T A C

Sticky ends from different BsrDI sites may not be compatible.
PflFI  (3004)
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  (3004)
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.
FspI  (2988)
1 site
T G C G C A A C G C G T
MscI  (2968)
1 site
T G G C C A A C C G G T
PluTI  (2889)
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  (2887)
1 site
G G C G C C C C G C G G
NarI  (2886)
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  (2885)
1 site
G G C G C C C C G C G G
BspDI  (2726)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (2726)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
StuI  (2707)
1 site
A G G C C T T C C G G A
SfiI  (2661)
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.
CsiI  (2475)
1 site
A C C W G G T T G G W C C A

Sticky ends from different CsiI sites may not be compatible.
SexAI  (2475)
1 site
A C C W G G T T G G W C C A
* Blocked by Dcm methylation.
Sticky ends from different SexAI sites may not be compatible.
AseI  (7)
1 site
A T T A A T T A A T T A
NdeI  (234)
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.
SnaBI  (340)
1 site
T A C G T A A T G C A T
NheI  (591)
1 site
G C T A G C C G A T C G
BmtI  (595)
1 site
G C T A G C C G A T C G
AfeI  (596)
1 site
A G C G C T T C G C G A
BglII  (609)
1 site
A G A T C T T C T A G A
PaeR7I  (613)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (613)
1 site
C T C G A G G A G C T C
Eco53kI  (618)
1 site
G A G C T C C T C G A G
SacI  (620)
1 site
G A G C T C C T C G A G
EcoRI  (629)
1 site
G A A T T C C T T A A G
SalI  (639)
1 site
G T C G A C C A G C T G
AccI  (640)
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.
Acc65I  (645)
1 site
G G T A C C C C A T G G
KpnI  (649)
1 site
G G T A C C C C A T G G
SacII  (652)
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.
PspOMI  (653)
1 site
G G G C C C C C C G G G
TspMI  (656)
1 site
C C C G G G G G G C C C
XmaI  (656)
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.
ApaI  (657)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
SmaI  (658)
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  (660)
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.
AgeI  (666)
1 site
A C C G G T T G G C C A
BsrGI  (1388)
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.
BlpI  (1399)
1 site
G C T