pYES-DEST52

Gateway® destination vector for galactose-inducible high-level expression of proteins in Saccharomyces cerevisiae.

Sequence Author: Thermo Fisher (Invitrogen)

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NaeI (7476) NgoMIV (7474) SwaI (7148) SnaBI (6844) HpaI (6199) PfoI * (6097) NruI (5793) BlpI (5757) BspDI - ClaI (5613) BmtI (5608) NheI (5604) NdeI (5464) BsgI (5435) SbfI (5401) XcmI (5252) EcoRV (5191) BsaHI (4231) BglI (3814) SpeI (7616) T7 promoter SacII (643) BspEI (980) EcoRI (984) PasI - PflMI * (1216) BssHII (1478) BbvCI (1709) TspMI - XmaI (1853) SmaI - SrfI (1855) BsaBI * (1868) BstXI (1972) SalI (2112) 6xHis PmeI (2340) BciVI (3004) BspHI (3521) AhdI (3694) pYES-DEST52 7621 bp
NaeI  (7476)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NaeI recognition sequence.
NgoMIV  (7474)
1 site
G C C G G C C G G C C G

Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
SwaI  (7148)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
SnaBI  (6844)
1 site
T A C G T A A T G C A T
HpaI  (6199)
1 site
G T T A A C C A A T T G
PfoI  (6097)
1 site
T C C N G G A A G G N C C T
* Blocked by Dcm methylation.
Sticky ends from different PfoI sites may not be compatible.
NruI  (5793)
1 site
T C G C G A A G C G C T
BlpI  (5757)
1 site
G C T N A G C C G A N T C G

Sticky ends from different BlpI sites may not be compatible.
BspDI  (5613)
1 site
A T C G A T T A G C T A
ClaI  (5613)
1 site
A T C G A T T A G C T A
BmtI  (5608)
1 site
G C T A G C C G A T C G
NheI  (5604)
1 site
G C T A G C C G A T C G
NdeI  (5464)
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.
BsgI  (5435)
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).
SbfI  (5401)
1 site
C C T G C A G G G G A C G T C C
XcmI  (5252)
1 site
C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C

The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
EcoRV  (5191)
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.
BsaHI  (4231)
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.
BglI  (3814)
1 site
G C C N N N N N G G C C G G N N N N N C C G

Sticky ends from different BglI sites may not be compatible.
SpeI  (7616)
1 site
A C T A G T T G A T C A
SacII  (643)
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.
BspEI  (980)
1 site
T C C G G A A G G C C T
EcoRI  (984)
1 site
G A A T T C C T T A A G
PasI  (1216)
1 site
C C C W G G G G G G W C C C

Sticky ends from different PasI sites may not be compatible.
PflMI  (1216)
1 site
C C A N N N N N T G G G G T N N N N N A C C
* Blocked by Dcm methylation.
Sticky ends from different PflMI sites may not be compatible.
BssHII  (1478)
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  (1709)
1 site
C C T C A G C G G A G T C G
TspMI  (1853)
1 site
C C C G G G G G G C C C
XmaI  (1853)
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.
SmaI  (1855)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SrfI  (1855)
1 site
G C C C G G G C C G G G C C C G
BsaBI  (1868)
1 site
G A T N N N N A T C C T A N N