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Plasmid Files

pET161-DEST

Gateway® destination vector for high-level bacterial expression of proteins with a C-terminal tetracysteine (Lumio™) tag.

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pET161-DEST.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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SgrAI (7440) SphI (7292) EcoNI (7227) BstEII (6577) ApaI (6556) PspOMI (6552) HpaI (6257) PshAI (5918) NruI (5660) FspAI (5178) PflFI - Tth111I (4413) BsaAI (4407) BspQI - SapI (4272) PciI (4155) BglII (1) XbaI (67) NdeI (107) NheI (112) BmtI (116) NotI (280) EcoRI (600) PasI (832) NcoI (901) BbvCI (1325) TspMI - XmaI (1469) SmaI - SrfI (1471) BmgBI (1505) SalI (1728) BstBI (1866) AgeI (1914) BlpI (1997) BspDI - ClaI (2241) ZraI (2346) AatII (2348) PvuI (2898) AhdI (3267) pET161-DEST 7480 bp
SgrAI  (7440)
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  (7292)
1 site
G C A T G C C G T A C G
EcoNI  (7227)
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.
BstEII  (6577)
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  (6556)
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  (6552)
1 site
G G G C C C C C C G G G
HpaI  (6257)
1 site
G T T A A C C A A T T G
PshAI  (5918)
1 site
G A C N N N N G T C C T G N N N N C A G

PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
NruI  (5660)
1 site
T C G C G A A G C G C T
FspAI  (5178)
1 site
R T G C G C A Y Y A C G C G T R
PflFI  (4413)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (4413)
1 site
G A C N N N G T C C T G N N N C A G

The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsaAI  (4407)
1 site
Y A C G T R R T G C A Y
BspQI  (4272)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (4272)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PciI  (4155)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BglII  (1)
1 site
A G A T C T T C T A G A
XbaI  (67)
1 site