Resources
Plasmid Files

pET-43.1 Ek/LIC

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

To see this sequence with restriction sites, features, and translations, please download
 SnapGene or the free  SnapGene Viewer.

pET-43.1 Ek_LIC Sequence and MappET-43.1 Ek_LIC.dna
Map and Sequence File   
Sequence Author:  Novagen (EMD Millipore)
Download Free Trial Get SnapGene Viewer

 DraIII (7053) PsiI (6925) AhdI (6641) BglI (6523) FspI (6418) ScaI (6160) AlwNI (5447) PciI (5031) BspQI - SapI (4915) PflFI - Tth111I (4776) Bpu10I (4137) PpuMI (4037) HpaI (3673) T7 terminator AvrII (483) PacI (503) 6xHis PaeR7I - XhoI (530) PmlI (576) EagI - NotI (592) HindIII (599) Acc65I (620) KpnI (624) SalI - SgrDI (626) PstI - SbfI (639) AscI (641) BsrGI (648) EcoRI (654) BamHI (660) Eco53kI (670) SacI (672) BseRI (699) PshAI (701) enterokinase site TspMI - XmaI (739) SmaI (741) S-Tag SacII (817) SpeI (848) BglII (963) BsmI (1007) MscI (1095) StuI (1148) BfuAI - BspMI (1297) NcoI (1423) NdeI (2337) RBS XbaI (2375) T7 promoter SphI (2642) EcoNI (2702) BstEII (3348) PspOMI (3374) ApaI (3378) pET-43.1 Ek/LIC 7295 bp
DraIII  (7053)
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  (6925)
1 site
T T A T A A A A T A T T
AhdI  (6641)
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.
BglI  (6523)
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  (6418)
1 site
T G C G C A A C G C G T
ScaI  (6160)
1 site
A G T A C T T C A T G A
AlwNI  (5447)
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  (5031)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BspQI  (4915)
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  (4915)
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.
PflFI  (4776)
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  (4776)
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.
Bpu10I  (4137)
1 site
C C T N A G C G G A N T C G

Efficient cleavage requires at least two copies of the Bpu10I
recognition sequence.
This recognition sequence is asymmetric, so ligating sticky ends
generated by Bpu10I will not always regenerate a Bpu10I site.
Sticky ends from different Bpu10I sites may not be compatible.
PpuMI  (4037)
1 site
R G G W C C Y Y C C W G G R

Sticky ends from different PpuMI sites may not be compatible.
HpaI  (3673)
1 site
G T T A A C C A A T T G
AvrII  (483)
1 site
C C T A G G G G A T C C
PacI  (503)
1 site
T T A A T T A A A A T T A A T T
PaeR7I  (530)
1 site
C T C G A G G A G C T C

PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (530)
1 site
C T C G A G G A G C T C
PmlI  (576)
1 site
C A C G T G G T G C A C

PmlI gradually loses activity when stored at -20°C.
EagI  (592)
1 site
C G G C C G G C C G G C
NotI  (592)
1 site
G C G G C C G C C G C C G G C G
HindIII  (599)
1 site
A A G C T T T T C G A A
Acc65I  (620)
1 site
G G T A C C C C A T G G
KpnI  (624)
1 site
G G T A C C C C A T G G
SalI  (626)
1 site
G T C G A C C A G C T G
SgrDI  (626)
1 site
C G T C G A C G G C A G C T G C
PstI  (639)
1 site
C T G C A G G A C G T C
SbfI  (639)
1 site
C C T G C A G G G G A C G T C C
AscI  (641)
1 site
G G C G C G C C C C G C G C G G
BsrGI  (648)
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  (654)
1 site
G A A T T C C T T A A G
BamHI  (660)
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.
Eco53kI  (670)
1 site
G A G C T C C T C G A G
SacI  (672)
1 site
G A G C T C C T C G A G
BseRI  (699)
1 site
G A G G A G ( N ) 8 N N C T C C T C ( N ) 8

Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the
DNA.
PshAI  (701)
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.
TspMI  (739)
1 site
C C C G G G G G G C C C
XmaI  (739)
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  (741)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
SacII  (817)
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  (848)
1 site
A C T A G T T G A T C A
BglII  (963)
1 site
A G A T C T T C T A G A
BsmI  (1007)
1 site
G A A T G C N C T T A C G N

Sticky ends from different BsmI sites may not be compatible.
MscI  (1095)
1 site
T G G C C A A C C G G T
StuI  (1148)
1 site
A G G C C T T C C G G A
BfuAI  (1297)
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  (1297)
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.
NcoI  (1423)
1 site
C C A T G G G G T A C C
NdeI  (2337)
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  (2375)
1 site
T C T A G A A G A T C T
SphI  (2642)
1 site
G C A T G C C G T A C G
EcoNI  (2702)
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  (3348)
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.
PspOMI  (3374)
1 site
G G G C C C C C C G G G
ApaI  (3378)
1 site
G G G C C C C C C G G G

ApaI can be used between 25°C and 37°C.
NusA
854 .. 2338  =  1485 bp
495 amino acids  =  54.9 kDa
Product: transcription elongation protein (N
utilization substance protein A)
highly soluble in E. coli
NusA
854 .. 2338  =  1485 bp
495 amino acids  =  54.9 kDa
Product: transcription elongation protein (N
utilization substance protein A)
highly soluble in E. coli
lacI
2817 .. 3899  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lacI
2817 .. 3899  =  1083 bp
360 amino acids  =  38.6 kDa
Product: lac repressor
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
AmpR
5854 .. 6714  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 1:  signal sequence  
   5854 .. 5922  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5854 .. 6714  =  861 bp
286 amino acids  =  31.6 kDa
   Segment 2:  
   5923 .. 6714  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
AmpR
5854 .. 6714  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and
related antibiotics
ori
5092 .. 5680  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
ori
5092 .. 5680  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin
of replication
f1 ori
6829 .. 7284  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
f1 ori
6829 .. 7284  =  456 bp
f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis
rop
4471 .. 4662  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
rop
4471 .. 4662  =  192 bp
63 amino acids  =  7.2 kDa
Product: Rop protein
MCS
530 .. 673  =  144 bp
multiple cloning site
MCS
530 .. 673  =  144 bp
multiple cloning site
rrnB T1 terminator
260 .. 346  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
rrnB T1 terminator
260 .. 346  =  87 bp
transcription terminator T1 from the E. coli rrnB
gene
lacI promoter
2739 .. 2816  =  78 bp
lacI promoter
2739 .. 2816  =  78 bp
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
T7 terminator
26 .. 73  =  48 bp
transcription terminator for bacteriophage T7 RNA
polymerase
S-Tag
767 .. 811  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
S-Tag
767 .. 811  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from
pancreatic ribonuclease A
lac operator
2383 .. 2407  =  25 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
2383 .. 2407  =  25 bp
The lac repressor binds to the lac operator to inhibit
transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-β-D-thiogalactopyranoside (IPTG).
T7 promoter
2408 .. 2426  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
2408 .. 2426  =  19 bp
promoter for bacteriophage T7 RNA polymerase
6xHis
512 .. 529  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
512 .. 529  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
thrombin site
734 .. 751  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
thrombin site
734 .. 751  =  18 bp
6 amino acids  =  627.8 Da
Product: thrombin recognition and cleavage site
6xHis
821 .. 838  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
821 .. 838  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
enterokinase site
701 .. 715  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
701 .. 715  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
RBS
2347 .. 2352  =  6 bp
ribosome binding site
RBS
2347 .. 2352  =  6 bp
ribosome binding site
HSV tag
536 .. 568  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
536 .. 568  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
Try SnapGene and create your own beautiful maps

Individual Sequences & Maps

SnapGene offers the fastest and easiest way to plan, visualize, and document your molecular biology procedures.

Priced accessibly so that everyone in your lab can have a license.

Learn More...

SnapGene Viewer is a versatile tool for creating and sharing richly annotated sequence files. It opens many common file formats.

Free! Because there should be no barriers to seeing your data.

Learn More...

The map, notes, and annotations on this page and in the sequence/map file are copyrighted material. This material may be used without restriction by academic, nonprofit, and governmental entities, except that the source must be cited as "www.snapgene.com/resources". Commercial entities must contact GSL Biotech LLC for permission and terms of use.

Copyright © 2016 GSL Biotech LLC | Site Map | Privacy | Legal Disclaimers   Subscribe to Our Newsletter