pET-53-DEST

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

Sequence Author: MilliporeSigma (Novagen)

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AvrII (6731) BstBI (6712) Strep-Tag II 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 (6749) 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-53-DEST™ 6832 bp
AvrII  (6731)
1 site
C C T A G G G G A T C C
BstBI  (6712)
1 site
T T C G A A A A G C T T
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 G G A G T C G
PasI  (5662)
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  (5430)
1 site
G A A T T C C T T A A G
EagI  (5110)
1 site
C G G C C G G C C G G C
NotI  (5110)
1 site
G C G G C C G C C G C C G G C G
NcoI  (4955)
1 site
C C A T G G G G T A C C
XbaI  (4916)
1 site
T C T A G A A G A T C T
BspDI  (4849)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (4849)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
SgrAI  (4805)
1 site
C R C C G G Y G G Y G G C C R C

Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SphI  (4657)
1 site
G C A T G C C G T A C G
EcoNI  (4592)
1 site
C C T N N N N N A G G G G A N N N N N T C C

The 1-base overhangs produced by EcoNI may be hard to ligate.
Sticky ends from different EcoNI sites may not be compatible.
BstAPI  (4448)
1 site
G C A N N N N N T G C C G T