pET-58-DEST

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

Sequence Author: MilliporeSigma (Novagen)

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AvrII (8294) AflII (8281) SalI (8088) PstI (8086) BmgBI (7865) SmaI - SrfI (7831) AvaI - BsoBI - TspMI - XmaI (7829) BbvCI (7685) PasI (7192) EcoRI (6960) lac UV5 promoter EagI - NotI (6640) PmlI (6502) thrombin site SacI (6492) Eco53kI (6490) BglII (6369) StuI (6188) AatII (5132) ZraI (5130) BstBI (5069) BmtI (5000) NheI (4996) NdeI (4991) 6xHis ATG RBS XbaI (4916) T7 promoter SphI (4657) EcoNI (4592) T7 terminator DraIII (245) PsiI (370) BsaI (716) BglI (775) FspI (877) PciI (2260) BspQI - SapI (2377) PflFI - Tth111I (2518) PpuMI (3255) HpaI (3622) PspOMI (3917) ApaI (3921) BstEII (3942) pET-58-DEST™ 8395 bp
AvrII  (8294)
1 site
C C T A G G G G A T C C
AflII  (8281)
1 site
C T T A A G G A A T T C
SalI  (8088)
1 site
G T C G A C C A G C T G
PstI  (8086)
1 site
C T G C A G G A C G T C
BmgBI  (7865)
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  (7831)
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  (7831)
1 site
G C C C G G G C C G G G C C C G
AvaI  (7829)
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  (7829)
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  (7829)
1 site
C C C G G G G G G C C C
XmaI  (7829)
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  (7685)
1 site
C C T C A G C G G A G T C G
PasI  (7192)
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  (6960)
1 site
G A A T T C C T T A A G
EagI  (6640)
1 site
C G G C C G G C C G G C
NotI  (6640)
1 site
G C G G C C G C C G C C G G C G
PmlI  (6502)
1 site
C A C G T G G T G C A C
SacI  (6492)
1 site
G A G C T C C T C G A G
Eco53kI  (6490)
1 site
G A G C T C C T C G A G
BglII  (6369)
1 site
A G A T C T T C T A G A
StuI  (6188)
1 site
A G G C C T T C C G G A
AatII  (5132)
1 site
G A C G T C C T G C A G
ZraI  (5130)
1 site
G A C G T C C T G C A G
BstBI  (5069)
1 site
T T C G A A A A G C T T
BmtI  (5000)
1 site
G C T A G C C G A T C G
NheI  (4996)
1 site
G C T A G C C G A T C G
NdeI  (4991)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
XbaI  (4916)
1 site
T C T A G A A G A T C T
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.
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
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
T G C G C A A C G C G