pcDNA6.2 EmGFP-Bsd V5-DEST

Vivid Colors™ Gateway® destination vector for expressing proteins in mammalian cells, with an EmGFP-Bsd fusion as a transfection marker.

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (8196) PciI (6382) BspQI - SapI (6266) EagI (5431) BbsI (5400) StuI (4316) PGK promoter MfeI (161) CMV enhancer NdeI (484) SnaBI (590) Eco53kI (816) SacI (818) PstI (1434) BfuAI - BspMI (1437) BstXI (1572) TspMI - XmaI (1683) SmaI - SrfI (1685) BbvCI (1828) BssHII (2058) BamHI (2099) EcoRI (2555) PspOMI (3188) ApaI (3192) SacII (3195) BstBI (3199) PmeI (3267) BstAPI (3533) PaeR7I - PspXI - XhoI (4011) SphI (4061) pcDNA™6.2/EmGFP-Bsd/V5-DEST 8198 bp
SgrDI  (8196)
1 site
C G T C G A C G G C A G C T G C
PciI  (6382)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (6266)
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  (6266)
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.
EagI  (5431)
1 site
C G G C C G G C C G G C
BbsI  (5400)
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.
StuI  (4316)
1 site
A G G C C T T C C G G A
MfeI  (161)
1 site
C A A T T G G T T A A C
NdeI  (484)
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  (590)
1 site
T A C G T A A T G C A T
Eco53kI  (816)
1 site
G A G C T C C T C G A G
SacI  (818)
1 site
G A G C T C C T C G A G
PstI  (1434)
1 site
C T G C A G G A C G T C
BfuAI  (1437)
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  (1437)
1 site
A C C T G C ( N ) 4 T G G A C G ( N )