pET-61-DEST

Gateway® Nova bacterial destination vector for expressing proteins tagged at the N-terminus with 6xHis and Strep-Tag II.

Sequence Author: MilliporeSigma (Novagen)

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AvrII (6745) SalI (6594) PstI (6592) BfuAI - BspMI (6581) BmgBI (6371) SmaI - SrfI (6337) AvaI - BsoBI - TspMI - XmaI (6335) BbvCI (6191) PasI (5698) EcoRI (5466) EagI - NotI (5146) BstBI (5003) Strep-Tag II ATG NcoI (4955) XbaI (4916) lac operator T7 promoter BspDI * - ClaI * (4849) SgrAI (4805) SphI (4657) EcoNI (4592) BstAPI (4448) BclI * (4110) BstEII (3942) ApaI (3921) PspOMI (3917) HpaI (3622) BlpI (6763) 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-61-DEST™ 6846 bp
AvrII  (6745)
1 site
C C T A G G G G A T C C
SalI  (6594)
1 site
G T C G A C C A G C T G
PstI  (6592)
1 site
C T G C A G G A C G T C
BfuAI  (6581)
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  (6581)
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.
BmgBI  (6371)
1 site
C A C G T C G T G C A G

This recognition sequence is asymmetric, so ligating blunt ends generated by BmgBI will not always regenerate a BmgBI site.
SmaI  (6337)
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  (6337)
1 site
G C C C G G G C C G G G C C C G
AvaI  (6335)
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  (6335)
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  (6335)
1 site
C C C G G G G G G C C C
XmaI  (6335)
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  (6191)
1 site
C C T C A G C G G A G T C G
PasI  (5698)
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.
EcoRI  (5466)
1 site
G A A T T C C T T A A G
EagI  (5146)
1 site
C G G C C G G C C G G C
NotI  (5146)
1 site
G C G G C C G C C G C C G G C G
BstBI  (5003)
1 site
T T C G A A A A G C T T
NcoI  (4955)
1 site
C C A T G G G G T A C C