pcDNA6.2 cLumio-DEST

Mammalian Gateway® destination vector for C-terminal tagging of a protein with the Lumio™ tetracysteine tag.

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (6807) AhdI (5886) PciI (4996) BspQI - SapI (4880) PfoI (4420) BsgI (4116) PpuMI (4088) BbsI (4014) BclI * (3851) AvrII (3800) StuI (3799) CsiI - SexAI * (3567) MfeI (161) SpeI (249) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) NotI (1041) lac UV5 promoter BspEI (1357) EcoRI (1361) BsmBI - Esp3I (1585) PflMI * (1593) BssHII (1855) BbvCI (2086) SrfI (2232) BmgBI (2266) BstXI (2349) BfuAI - BspMI (2476) PstI (2487) HpaI (2629) AgeI (2678) tetracysteine tag PmeI (2732) pcDNA™6.2/cLumio™-DEST 6809 bp
SgrDI  (6807)
1 site
C G T C G A C G G C A G C T G C
AhdI  (5886)
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.
PciI  (4996)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4880)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (4880)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PfoI  (4420)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
BsgI  (4116)
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).
PpuMI  (4088)
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.
BbsI  (4014)
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.
BclI  (3851)
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.
AvrII  (3800)
1 site
C C T A G G G G A T C C
StuI  (3799)
1 site
A G G C C T T C C G G A
CsiI  (3567)
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  (3567)
1 site
A C C W G G T T G