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pET-41 Ek/LIC (linearized)

Linearized bacterial vector for ligation-independent cloning (LIC) to express GST-tagged proteins with an enterokinase site.

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pET-41 Ek_LIC (linearized).dna
Map and Sequence File:    Download    Open   
Sequence Author:  Novagen (EMD Millipore)
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DraIII (5428) PsiI (5300) AsiSI - PvuI (4727) SmaI (4601) TspMI - XmaI (4599) NruI (4384) AcuI (4073) AlwNI (3941) BssS α I (3698) BstZ17I (3296) PflFI - Tth111I (3270) BlpI (5750) AvrII (5801) PaeR7I - PspXI - XhoI (5844) EagI - NotI (5852) HindIII (5859) SalI (5865) Eco53kI (5874) SacI (5876) PstI - SbfI (5884) AscI (5886) StuI (5896) BsrGI (5899) EcoRI (5905) BamHI (5911) EcoRV (5921) NcoI (5924) End (5931) Start (0) AgeI (29) Acc65I (32) KpnI (36) BglII (39) MfeI (97) SacII (130) SpeI (161) SwaI (404) MscI (624) NdeI (831) XbaI (869) SgrAI (980) SphI (1136) BstAPI (1344) MluI (1661) BstEII (1842) NmeAIII (1867) PspOMI (1868) ApaI (1872) HpaI (2167) PpuMI (2531) pET-41 Ek/LIC 5931 bp
DraIII  (5428)
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  (5300)
1 site
T T A T A A A A T A T T
AsiSI  (4727)
1 site
G C G A T C G C C G C T A G C G
PvuI  (4727)
1 site
C G A T C G G C T A G C
SmaI  (4601)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
TspMI  (4599)
1 site
C C C G G G G G G C C C
XmaI  (4599)
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.
NruI  (4384)
1 site
T C G C G A A G C G C T
AcuI  (4073)
1 site
C T G A A G ( N ) 14 N N G A C T T C ( N ) 14

Efficient cleavage requires at least two copies of the AcuI recognition sequence.
Sticky ends from different AcuI sites may not be compatible.
After cleavage, AcuI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
AlwNI  (3941)
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.
BssSαI  (3698)
1 site
C A C G A G G T G C T C
BstZ17I  (3296)
1 site
G T A T A C C A T A T G
PflFI  (3270)
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  (3270)
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  (5750)
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.
AvrII  (5801)
1 site
C C T A G G G G A T C C
PaeR7I  (5844)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (5844)
1 site
V C T C G A G B B G A G C T C V
XhoI  (5844)
1 site
C T C G A G G A G C T C
EagI  (5852)
1 site
C G G C C G G C C G G C
NotI  (5852)
1 site
G C G G C C G C C G C C G G C G
HindIII  (5859)
1 site
A A G C T T T T C G A A
SalI  (5865)
1 site
G T C G A C C A G C T G
Eco53kI  (5874)
1 site
G A G C T C C T C G A G
SacI  (5876)
1 site
G A G C T C C T C G A G
PstI  (5884)
1 site
C T G C A G G A C G T C
SbfI  (5884)
1 site
C C T G C A G G G G A C G T C C
AscI  (5886)
1 site
G G C G C G C C C C G C G C G G
StuI  (5896)
1 site
A G G C C T T C C G G A
BsrGI  (5899)
1 site
T G T A C A A C A T G T

BsrGI is typically used at 37°C, but is even more active at 60°C.
EcoRI  (5905)
1 site
G A A T T C C T T A A G
BamHI  (5911)
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  (5921)
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  (5924)
1 site
C C A T G G G G T A C C
End  (5931)
0 sites
Start  (0)
0 sites
AgeI  (29)
1 site
A C C G G T T G G C C A
Acc65I  (32)
1 site
G G T A C C C C A T G G
KpnI  (36)
1 site
G G T A C C C C A T G G
BglII  (39)
1 site
A G A T C T T C T A G A
MfeI  (97)
1 site
C A A T T G G T T A A C
SacII  (130)
1 site
C C G C G G G G C G C C

Efficient cleavage requires at least two copies of the SacII recognition sequence.
SpeI  (161)
1 site
A C T A G T T G A T C A
SwaI  (404)
1 site
A T T T A A A T T A A A T T T A

SwaI is typically used at 25°C, but is 50% active at 37°C.
MscI  (624)
1 site
T G G C C A A C C G G T
NdeI  (831)
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  (869)
1 site
T C T A G A A G A T C T
SgrAI  (980)
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  (1136)
1 site