pcDNA6.2 cGeneBLAzer-DEST

Gateway® destination vector for in vivo or in vitro detection of mammalian gene expression using a C-terminal β-lactamase reporter.

Sequence Author: Thermo Fisher (Invitrogen)

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SgrDI (7517) PciI (5706) BspQI - SapI (5590) PfoI (5130) BsgI (4826) PpuMI (4798) BbsI (4724) BclI * (4561) AvrII (4510) StuI (4509) CsiI - SexAI * (4277) MfeI (161) MluI (228) 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) BstEII (3420) AgeI (3424) PmeI (3442) pcDNA™6.2/cGeneBLAzer™-DEST 7519 bp
SgrDI  (7517)
1 site
C G T C G A C G G C A G C T G C
PciI  (5706)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (5590)
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  (5590)
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  (5130)
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  (4826)
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  (4798)
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  (4724)
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  (4561)
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  (4510)
1 site
C C T A G G G G A T C C
StuI  (4509)
1 site
A G G C C T T C C G G A
CsiI  (4277)
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  (4277)
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.
MfeI  (161)
1 site
C A A T T G G T T A A C
MluI  (228)
1 site
A C G C G T T G C G C A
SpeI  (249)
1 site
A C T A G T T G A T C A
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