pIEx-2

Vector for high-level expression in insect cells of proteins with a cleavable N-terminal GST-6xHis-S-Tag cassette plus a C-terminal HSV tag.

Sequence Author: MilliporeSigma (Novagen)

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HomePlasmidsInsect Cell VectorspIEx-2
DrdI (4095) PspFI (3897) BseYI (3893) AhdI (3309) BsaI (3243) BpmI (3240) NmeAIII (3162) AatII (2390) ZraI (2388) XbaI (2382) PflFI - Tth111I (2327) BspDI * - ClaI * (207) EcoRV (239) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BsiWI (784) MluI (872) BtgI - NcoI (1082) EcoNI (1097) EcoO109I (1119) MscI (1294) BsgI (1373) SwaI (1514) SpeI (1753) 6xHis XcmI (1786) PflMI (1830) thrombin site PshAI (1904) BseRI (1908) BamHI (1926) MfeI (1932) Eco53kI (1941) BanII - SacI (1943) BsrGI (1946) AscI (1954) PstI - SbfI (1964) SalI (1966) Acc65I (1972) AgeI (1975) KpnI (1976) HindIII (1984) NotI (1991) BstZ17I (2006) PmlI (2011) BbsI (2045) PaeR7I - XhoI (2053) DraIII (2064) Bsu36I (2074) PacI (2103) IE1 terminator pIEx™-2 4563 bp
DrdI  (4095)
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  (3897)
1 site		 C C C A G C G G G T C G
BseYI  (3893)
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.
AhdI  (3309)
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  (3243)
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.
BpmI  (3240)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
NmeAIII  (3162)
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).
AatII  (2390)
1 site		 G A C G T C C T G C A G
ZraI  (2388)
1 site		 G A C G T C C T G C A G
XbaI  (2382)
1 site		 T C T A G A A G A T C T
PflFI  (2327)
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  (2327)
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.
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.
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
EcoNI  (1097)
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.
EcoO109I  (1119)
1 site		 R G G N C C Y Y C C N G G R
Sticky ends from different EcoO109I sites may not be compatible.
MscI  (1294)
1 site		 T G G C C A A C C G G T
BsgI  (1373)
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).
SwaI  (1514)
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.
SpeI  (1753)
1 site		 A C T A G T T G A T C A
XcmI  (1786)
1 site		 C C A N N N N N N N N N T G G G G T N N N N N N N N N A C C
The 1-base overhangs produced by XcmI may be hard to ligate.
Sticky ends from different XcmI sites may not be compatible.
PflMI  (1830)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
PshAI  (1904)
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.
BseRI  (1908)
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.
BamHI  (1926)
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.
MfeI  (1932)
1 site		 C A A T T G G T T A A C
Eco53kI  (1941)
1 site		 G A G C T C C T C G A G
BanII  (1943)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
SacI  (1943)
1 site		 G A G C T C C T C G A G
BsrGI  (1946)
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.
AscI  (1954)
1 site		 G G C G C G C C C C G C G C G G
PstI  (1964)
1 site		 C T G C A G G A C G T C
SbfI  (1964)
1 site		 C C T G C A G G G G A C G T C C
SalI  (1966)
1 site		 G T C G A C C A G C T G
Acc65I  (1972)
1 site		 G G T A C C C C A T G G
AgeI  (1975)
1 site		 A C C G G T T G G C C A
KpnI  (1976)
1 site		 G G T A C C C C A T G G
HindIII  (1984)
1 site		 A A G C T T T T C G A A
NotI  (1991)
1 site		 G C G G C C G C C G C C G G C G
BstZ17I  (2006)
1 site		 G T A T A C C A T A T G
PmlI  (2011)
1 site		 C A C G T G G T G C A C
BbsI  (2045)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PaeR7I  (2053)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2053)
1 site		 C T C G A G G A G C T C
DraIII  (2064)
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  (2074)
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  (2103)
1 site		 T T A A T T A A A A T T A A T T
GST
1084 .. 1740  =  657 bp
219 amino acids  =  25.6 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
1084 .. 1740  =  657 bp
219 amino acids  =  25.6 kDa
Product: glutathione S-transferase from Schistosoma japonicum
6xHis
1768 .. 1785  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1768 .. 1785  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
S-Tag
1795 .. 1839  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
1795 .. 1839  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
thrombin site
1855 .. 1872  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
1855 .. 1872  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
enterokinase site
1891 .. 1905  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
1891 .. 1905  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
HSV tag
2020 .. 2052  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
2020 .. 2052  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
AmpR
2522 .. 3382  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   2522 .. 2590  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2522 .. 3382  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   2591 .. 3382  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
2522 .. 3382  =  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
3553 .. 4141  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
3553 .. 4141  =  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
2080 .. 2387  =  308 bp
terminator of the ie1 gene from the baculovirus Autographa californica
IE1 terminator
2080 .. 2387  =  308 bp
terminator of the ie1 gene from the baculovirus Autographa californica
AmpR promoter
2417 .. 2521  =  105 bp
AmpR promoter
2417 .. 2521  =  105 bp
MCS
1900 .. 2014  =  115 bp
multiple cloning site
MCS
1900 .. 2014  =  115 bp
multiple cloning site
ORF:  1084 .. 2064  =  981 bp
ORF:  326 amino acids  =  36.9 kDa
ORF:  1745 .. 2068  =  324 bp
ORF:  107 amino acids  =  12.0 kDa
ORF:  2522 .. 3382  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2986 .. 3252  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  452 .. 1090  =  639 bp
ORF:  212 amino acids  =  24.2 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

Plasmid Sets

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pET & Duet Vectors (Novagen)
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Coronavirus Resources
Insect Cell Vectors
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TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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