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) lac UV5 promoter 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 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 N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
BclI  (4110)
1 site
T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstEII  (3942)
1 site
G G T N A C C C C A N T G G

Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
ApaI  (3921)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
PspOMI  (3917)
1 site
G G G C C C C C C G G G
HpaI  (3622)
1 site
G T T A A C C A A T T G
BlpI  (6782)
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.
DraIII  (245)
1 site
C A C N N N G T G G T G N N N C A C

Sticky ends from different DraIII sites may not be compatible.
PsiI  (370)
1 site
T T A T A A A A T A T T
AhdI  (655)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BsaI  (716)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BglI  (775)
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.
FspI  (877)
1 site