pIEx-8 Ek_LIC

Insect cell vector for ligation-independent cloning (LIC) to express proteins with an N-terminal Strep-Tag® II-enterokinase cassette and a C-terminal 10xHis tag.

Sequence Author: MilliporeSigma (Novagen)

|Download SnapGene Viewer
Explore Over 2.7k Plasmids: Insect Cell Vectors | More Plasmid Sets
No matches
AvaI - BsoBI - TspMI - XmaI (3733) BspQI - SapI (3489) PciI (3372) DrdI (3270) PspFI (3072) BseYI (3068) AlwNI (2963) AhdI (2484) BsaI (2418) BsrFI (2399) NmeAIII (2337) ScaI (2003) SmaI (3735) BspDI * - ClaI * (207) EcoRV (239) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BclI * (778) BsiWI (784) MluI (872) BtgI - NcoI (1082) ATG BstBI (1109) Acc65I (1137) BsgI (1139) KpnI (1141) BamHI (1143) PshAI (1153) SalI (1156) AccI (1157) HindIII (1162) NotI (1169) Eco53kI (1180) BanII - BseRI - SacI (1182) DraIII (1239) Bsu36I (1249) PacI (1278) PflFI - Tth111I (1502) BtgZI (1525) XbaI (1557) XmnI (1884) pIEx™-8 Ek/LIC 3738 bp
AvaI  (3733)
1 site
C Y C G R G G R G C Y C

Sticky ends from different AvaI sites may not be compatible.
BsoBI  (3733)
1 site
C Y C G R G G R G C Y C

Sticky ends from different BsoBI sites may not be compatible.
BsoBI is typically used at 37°C, but can be used at temperatures up to 65°C.
TspMI  (3733)
1 site
C C C G G G G G G C C C
XmaI  (3733)
1 site
C C C G G G G G G C C C

Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
BspQI  (3489)
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  (3489)
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  (3372)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
DrdI  (3270)
1 site
G A C N N N N N N G T C C T G N N N N N N C A G

Sticky ends from different DrdI sites may not be compatible.
PspFI  (3072)
1 site
C C C A G C G G G T C G
BseYI  (3068)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (2963)
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.
AhdI  (2484)
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.
BsaI  (2418)
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.
BsrFI  (2399)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
NmeAIII  (2337)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
ScaI  (2003)
1 site
A G T A C T T C A T G A
SmaI  (3735)
1 site
C C C G G G G G G C C C

SmaI can be used at 37°C for brief incubations.
BspDI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
ClaI  (207)
1 site
A T C G A T T A G C T A
* Blocked by Dam methylation.
EcoRV  (239)
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.
PsiI  (409)
1 site
T T A T A A A A T A T T
NheI  (606)
1 site
G C T A G C C G A T C G
BmtI  (610)
1 site
G C T A G C C G A T C G
SphI  (614)
1 site
G C A T G C C G T A C G
BsmI  (775)
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.
BclI  (778)
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.
BsiWI  (784)
1 site
C G T A C G G C A T G C

BsiWI is typically used at 55°C, but is 50% active at 37°C.
MluI  (872)
1 site
A C G C G T T G C G C A
BtgI  (1082)
1 site
C C R Y G G G G Y R C C

Sticky ends from different BtgI sites may not be compatible.
NcoI  (1082)
1 site
C C A T G G G G T A C C
BstBI  (1109)
1 site
T T C G A A A A G C T T
Acc65I  (1137)
1 site
G G T A C C C C A T G G
BsgI  (1139)
1 site
G T G C A G ( N ) 14 N N C A C G T C ( N ) 14

Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
KpnI  (1141)
1 site
G G T A C C C C A T G G
BamHI  (1143)
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.
PshAI  (1153)
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.
SalI  (1156)
1 site
G T C G A C C A G C T G
AccI  (1157)
1 site
G T M K A C C A K M T G

Efficient cleavage with AccI requires ≥13 bp on each side of the recognition sequence.
Sticky ends from different AccI sites may not be compatible.
HindIII  (1162)
1 site
A A G C T T T T C G A A
NotI  (1169)
1 site
G C G G C C G C C G C C G G C G
Eco53kI  (1180)
1 site
G A G C T C C T C G A G
BanII  (1182)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
BseRI  (1182)
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.
SacI  (1182)
1 site
G A G C T C C T C G A G
DraIII  (1239)
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.
Bsu36I  (1249)
1 site
C C T N A G G G G A N T C C

Sticky ends from different Bsu36I sites may not be compatible.
PacI  (1278)
1 site
T T A A T T A A A A T T A A T T
PflFI  (1502)
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  (1502)
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.
BtgZI  (1525)
1 site
G C G A T G ( N ) 10 C G C T A C ( N ) 10 ( N ) 4

Sticky ends from different BtgZI sites may not be compatible.
After cleavage, BtgZI can remain bound to DNA and alter its electrophoretic mobility.
BtgZI is typically used at 60°C, but is 75% active at 37°C.
XbaI  (1557)
1 site
T C T A G A A G A T C T
XmnI  (1884)
1 site
G A A N N N N T T C C T T N N N N A A G
AmpR
1697 .. 2557  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1697 .. 1765  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1697 .. 2557  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1766 .. 2557  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1697 .. 2557  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
IE1 promoter
487 .. 1078  =  592 bp
promoter of the ie1 gene from the baculovirus Autographa californica
IE1 promoter
487 .. 1078  =  592 bp
promoter of the ie1 gene from the baculovirus Autographa californica
ori
2728 .. 3316  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2728 .. 3316  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
hr5 enhancer
1 .. 483  =  483 bp
baculovirus early transcription enhancer
hr5 enhancer
1 .. 483  =  483 bp
baculovirus early transcription enhancer
IE1 terminator
1255 .. 1562  =  308 bp
terminator of the ie1 gene from the baculovirus Autographa californica
IE1 terminator
1255 .. 1562  =  308 bp
terminator of the ie1 gene from the baculovirus Autographa californica
ATG
1084 .. 1086  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
1084 .. 1086  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
Strep-Tag II
1093 .. 1116  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
Strep-Tag II
1093 .. 1116  =  24 bp
8 amino acids  =  1.1 kDa
Product: peptide that binds Strep-Tactin®, an engineered form of streptavidin
enterokinase site
1123 .. 1137  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
1123 .. 1137  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
10xHis
1207 .. 1236  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
10xHis
1207 .. 1236  =  30 bp
10 amino acids  =  1.4 kDa
Product: 10xHis affinity tag
AmpR promoter
1592 .. 1696  =  105 bp
AmpR promoter
1592 .. 1696  =  105 bp
ORF:  1697 .. 2557  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2161 .. 2427  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  452 .. 1018  =  567 bp
ORF:  188 amino acids  =  21.5 kDa
Click here to try SnapGene

Download pIEx-8 Ek_LIC.dna file

SnapGene

SnapGene is the easiest way to plan, visualize and document your everyday molecular biology procedures

  • Fast accurate construct design for all major molecular cloning techniques
  • Validate sequenced constructs using powerful alignment tools
  • Customize plasmid maps with flexible annotation and visualization controls
  • Automatically generate a rich graphical history of every edit and procedure

SnapGene Viewer

SnapGene Viewer is free software that allows molecular biologists to create, browse, and share richly annotated sequence files.

  • Gain unparalleled visibility of your plasmids, DNA and protein sequences
  • Annotate features on your plasmids using the curated feature database
  • Store, search, and share your sequences, files and maps

Individual Sequences & Maps

The maps, notes, and annotations in the zip file on this page 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.

Discover the most user-friendly molecular biology experience.