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pET160-DEST

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

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pET160-DEST.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Thermo Fisher (Invitrogen)
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SphI (7249) EcoNI (7184) BclI * (6702) BstEII (6534) ApaI (6513) PspOMI (6509) HpaI (6214) PshAI (5875) NruI (5617) FspAI (5135) PflFI - Tth111I (4370) BsaAI (4364) BspQI - SapI (4229) PciI (4112) BglII (1) XbaI (67) NdeI (107) NsiI (113) AleI (128) NotI (322) EcoRI (642) PasI (874) AgeI (1142) BbvCI (1346) TspMI - XmaI (1490) SmaI - SrfI (1492) BmgBI (1526) SalI (1749) PacI (1889) BlpI (1950) BspDI - ClaI (2198) ZraI (2303) AatII (2305) PvuI (2855) AhdI (3224) pET160-DEST 7437 bp
SphI  (7249)
1 site
G C A T G C C G T A C G
EcoNI  (7184)
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.
BclI  (6702)
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  (6534)
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  (6513)
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  (6509)
1 site
G G G C C C C C C G G G
HpaI  (6214)
1 site
G T T A A C C A A T T G
PshAI  (5875)
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  (5617)
1 site
T C G C G A A G C G C T
FspAI  (5135)
1 site
R T G C G C A Y Y A C G C G T R
PflFI  (4370)
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  (4370)
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  (4364)
1 site
Y A C G T R R T G C A Y
BspQI  (4229)
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  (4229)
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  (4112)
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
T C T A G A A G A T C T
NdeI  (107)
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.
NsiI  (113)
1 site
A T G C A T T A C G T A
AleI  (128)
1 site
C A C N N N N G T G G T G N N N N C A C
NotI  (322)
1 site
G C G G C C G C C G C C G G C G
EcoRI  (642)
1 site
G A A T T C C T T A A G
PasI  (874)
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.
AgeI  (1142)
1 site
A C C G G T T G G C C A
BbvCI  (1346)
1 site
C C T C A G C G G A G T C G
TspMI  (1490)
1 site
C C C G G G G G G C C C
XmaI  (1490)
1 site
C C C G G G G G G C C C

Efficient cleavage requires at least two copies of the XmaI recognition sequence.
Full cleavage with XmaI may require a long incubation.
SmaI  (1492)
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  (1492)
1 site
G C C C G G G C C G G G C C C G
BmgBI  (1526)
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.
SalI  (1749)
1 site
G T C G A C C A G C T G
PacI  (1889)
1 site
T T A A T T A A A A T T A A T T
BlpI  (1950)
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.
BspDI  (2198)
1 site
A T C G A T T A G C T A
ClaI  (2198)
1 site