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pET-32 Ek/LIC

Bacterial vector for ligation-independent cloning (LIC) to express thioredoxin-tagged proteins with an enterokinase site.

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pET-32 Ek_LIC.dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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DraIII (5675) PsiI (5547) ScaI (5012) PvuI (4902) PstI (4777) BsaI (4593) AhdI (4532) AlwNI (4055) PciI (3639) BspQI - SapI (3523) BstZ17I (3410) PflFI - Tth111I (3384) BlpI (80) PaeR7I - PspXI - XhoI (158) EagI - NotI (166) HindIII (173) SalI (179) Eco53kI (188) SacI (190) EcoRI (192) BamHI (198) EcoRV (206) NcoI (211) TspMI - XmaI (228) SmaI - SrfI (230) BseRI (234) Acc65I (251) KpnI (255) BglII (258) BstBI (285) MscI (368) RsrII (606) XbaI (746) SgrAI (857) SphI (1013) EcoNI (1073) BstAPI (1221) MluI (1538) BclI * (1552) BstEII (1719) PspOMI (1745) ApaI (1749) BssHII (1949) HpaI (2044) PshAI (2383) FspAI (2620) PpuMI (2645) Bpu10I (2745) pET-32 Ek/LIC 5917 bp
DraIII  (5675)
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  (5547)
1 site
T T A T A A A A T A T T
ScaI  (5012)
1 site
A G T A C T T C A T G A
PvuI  (4902)
1 site
C G A T C G G C T A G C
PstI  (4777)
1 site
C T G C A G G A C G T C
BsaI  (4593)
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.
AhdI  (4532)
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.
AlwNI  (4055)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
PciI  (3639)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (3523)
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  (3523)
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.
BstZ17I  (3410)
1 site
G T A T A C C A T A T G
PflFI  (3384)
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  (3384)
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.
BlpI  (80)
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.
PaeR7I  (158)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (158)
1 site
V C T C G A G B B G A G C T C V
XhoI  (158)
1 site
C T C G A G G A G C T C
EagI  (166)
1 site
C G G C C G G C C G G C
NotI  (166)
1 site
G C G G C C G C C G C C G G C G
HindIII  (173)
1 site
A A G C T T T T C G A A
SalI  (179)
1 site
G T C G A C C A G C T G
Eco53kI  (188)
1 site
G A G C T C C T C G A G
SacI  (190)
1 site
G A G C T C C T C G A G
EcoRI  (192)
1 site
G A A T T C C T T A A G
BamHI  (198)
1 site
G G A T C C C C T A G G

After cleavage, BamHI-HF™ (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
EcoRV  (206)
1 site
G A T A T C C T A T A G

EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
NcoI  (211)
1 site
C