pET-62-DEST

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

Sequence Author: MilliporeSigma (Novagen)

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AvrII (6727) PacI (6723) PaeR7I - PspXI - XhoI (6711) BstBI (6692) 6xHis SalI (6520) PstI (6518) BfuAI - BspMI (6507) BmgBI (6297) SmaI - SrfI (6263) TspMI - XmaI (6261) BbvCI (6117) PasI (5624) EcoRI (5392) lac UV5 promoter EagI - NotI (5072) XbaI (4916) T7 promoter BspDI * - ClaI * (4849) SgrAI (4805) BtgI (4687) SphI (4657) EcoNI (4592) BstAPI (4448) BclI * (4110) BstEII (3942) ApaI (3921) PspOMI (3917) HpaI (3622) BlpI (6745) 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-62-DEST™ 6828 bp
AvrII  (6727)
1 site
C C T A G G G G A T C C
PacI  (6723)
1 site
T T A A T T A A A A T T A A T T
PaeR7I  (6711)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (6711)
1 site
V C T C G A G B B G A G C T C V
XhoI  (6711)
1 site
C T C G A G G A G C T C
BstBI  (6692)
1 site
T T C G A A A A G C T T
SalI  (6520)
1 site
G T C G A C C A G C T G
PstI  (6518)
1 site
C T G C A G G A C G T C
BfuAI  (6507)
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  (6507)
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  (6297)
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  (6263)
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  (6263)
1 site
G C C C G G G C C G G G C C C G
TspMI  (6261)
1 site
C C C G G G G G G C C C
XmaI  (6261)
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  (6117)
1 site
C C T C A G C G G A G T C G
PasI  (5624)
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  (5392)
1 site
G A A T T C C T T A A G
EagI  (5072)
1 site
C G G C C G G C C G G C
NotI  (5072)
1 site
G C G G C C G C C G C C G G C G
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.
BtgI  (4687)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
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  (6745)
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
T G C