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pDEST™8

Gateway® destination vector for expressing proteins in insect cells using the Bac-to-Bac® baculovirus system.

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pDEST8.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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BbsI (6403) PflFI - Tth111I (5874) BseRI (5607) EcoRV (5471) SacII (5414) BspQI - SapI (5027) PspFI (4610) BseYI (4606) FspI (3799) PvuI (3653) BsaHI (3482) SnaBI (1) NotI (286) PvuII (647) EcoRI (747) NcoI (1048) MluI (1194) BamHI (1200) BssHII (1241) BbvCI (1472) TspMI - XmaI (1616) SmaI - SrfI (1618) BmgBI (1652) BfuAI - BspMI (1862) PstI (1873) SalI (1875) MfeI (2123) HpaI (2136) BclI * (2272) AvrII (2287) NgoMIV (2773) NaeI (2775) BanII (2807) pDEST™8 6526 bp
BbsI  (6403)
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.
PflFI  (5874)
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  (5874)
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.
BseRI  (5607)
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.
EcoRV  (5471)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
SacII  (5414)
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.
BspQI  (5027)
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  (5027)
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.
PspFI  (4610)
1 site
C C C A G C G G G T C G
BseYI  (4606)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
FspI  (3799)
1 site
T G C G C A A C G C G T
PvuI  (3653)
1 site
C G A T C G G C T A G C
BsaHI  (3482)
1 site
G R C G Y C C Y G C R G

BsaHI is typically used at 37°C, but is even more active at 60°C.
SnaBI  (1)
1 site
T A C G T A A T G C A T
NotI  (286)
1 site
G C G G C C G C C G C C G G C G
PvuII  (647)
1 site
C A G C T G G T C G A C
EcoRI  (747)
1 site
G A A T T C C T T A A G
NcoI  (1048)
1 site
C C A T G G G G T A C C
MluI  (1194)
1 site
A C G C G T T G C G C A
BamHI  (1200)
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.
BssHII  (1241)
1 site
G C G C G C C G C G C G

BssHII is typically used at 50°C, but is 75% active at 37°C.
BbvCI  (1472)
1 site
C C T C A G C G G A G T C G
TspMI  (1616)
1 site
C C C G G G G G<