pET-60-DEST

Gateway® Nova bacterial destination vector for expressing proteins tagged at the N-terminus with GST.

Sequence Author: MilliporeSigma (Novagen)

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AvrII (7379) 6xHis SalI (7218) PstI (7216) BfuAI - BspMI (7205) BmgBI (6995) SmaI - SrfI (6961) AvaI - BsoBI - TspMI - XmaI (6959) BbvCI (6815) PasI (6322) EcoRI (6090) EagI - NotI (5770) PmlI (5632) thrombin site SwaI (5384) BstBI (5354) RBS XbaI (4916) T7 promoter BspDI * - ClaI * (4849) SgrAI (4805) BtgI (4687) SphI (4657) BstAPI (4448) BstEII (3942) ApaI (3921) PspOMI (3917) BlpI (7397) PsiI (370) AhdI (655) BsaI (716) BglI (775) FspI (877) PvuI (1025) PflFI - Tth111I (2518) PpuMI (3255) HpaI (3622) pET-60-DEST™ 7480 bp
AvrII  (7379)
1 site
C C T A G G G G A T C C
SalI  (7218)
1 site
G T C G A C C A G C T G
PstI  (7216)
1 site
C T G C A G G A C G T C
BfuAI  (7205)
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  (7205)
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  (6995)
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  (6961)
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  (6961)
1 site
G C C C G G G C C G G G C C C G
AvaI  (6959)
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  (6959)
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  (6959)
1 site
C C C G G G G G G C C C
XmaI  (6959)
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  (6815)
1 site
C C T C A G C G G A G T C G
PasI  (6322)
1 site
C C C W G G G G G G W C C C