pET-54-DEST

Gateway® Nova bacterial destination vector for expressing proteins tagged at the C-terminus with S-Tag.

Sequence Author: MilliporeSigma (Novagen)

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AvrII (6764) AflII (6751) SalI (6558) PstI (6556) BfuAI - BspMI (6545) SmaI - SrfI (6301) AvaI - BsoBI - TspMI - XmaI (6299) BbvCI (6155) PasI (5662) EcoRI (5430) EagI - NotI (5110) 6xHis ATG NcoI (4955) RBS XbaI (4916) T7 promoter BspDI * - ClaI * (4849) SgrAI (4805) SphI (4657) EcoNI (4592) BstAPI (4448) BclI * (4110) BstEII (3942) ApaI (3921) PspOMI (3917) HpaI (3622) BlpI (6782) DraIII (245) PsiI (370) AhdI (655) BsaI (716) BglI (775) FspI (877) PvuI (1025) PciI (2260) BspQI - SapI (2377) PflFI - Tth111I (2518) PpuMI (3255) pET-54-DEST™ 6865 bp
AvrII  (6764)
1 site
C C T A G G G G A T C C
AflII  (6751)
1 site
C T T A A G G A A T T C
SalI  (6558)
1 site
G T C G A C C A G C T G
PstI  (6556)
1 site
C T G C A G G A C G T C
BfuAI  (6545)
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  (6545)
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 BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
SmaI  (6301)
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  (6301)
1 site
G C C C G G G C C G G G C C C G
AvaI  (6299)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (6299)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (6299)
1 site
C C C G G G G G G C C C
XmaI  (6299)
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.
BbvCI  (6155)
1 site
C C T C A G C