pBiEx-2

Vector for high-level expression in both bacterial and 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 VectorspBiEx-2
DrdI (5694) AhdI (4908) BsaI (4842) PfoI (3883) BstAPI (3775) BstEII (3269) ApaI (3248) PspOMI (3244) BspDI * - ClaI * (207) PsiI (409) NheI (606) BmtI (610) SphI (614) BsmI (775) BseRI (781) BsiWI (784) SacII (1085) T7 promoter MscI (1389) SwaI (1609) SpeI (1848) 6xHis enterokinase site BglII (2008) SrfI (2020) BamHI (2027) MfeI (2033) Eco53kI (2042) SacI (2044) BsrGI (2047) AscI (2055) PstI - SbfI (2065) SalI (2067) Acc65I (2073) AgeI (2076) KpnI (2077) HindIII (2085) NotI (2092) BstZ17I (2107) PmlI (2112) PaeR7I - XhoI (2154) DraIII (2165) BbvCI - Bpu10I (2175) BlpI (2212) Bsu36I (2289) PacI (2318) PflFI - Tth111I (2542) KasI (2812) NarI * (2813) SfoI (2814) PluTI (2816) BsmBI - Esp3I (2836) HpaI (2949) pBiEx™-2 6162 bp
DrdI  (5694)
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.
AhdI  (4908)
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  (4842)
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.
PfoI  (3883)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
BstAPI  (3775)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
BstEII  (3269)
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.
ApaI  (3248)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
PspOMI  (3244)
1 site		 G G G C C C C C C G G G
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.
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.
BseRI  (781)
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.
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.
SacII  (1085)
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.
MscI  (1389)
1 site		 T G G C C A A C C G G T
SwaI  (1609)
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  (1848)
1 site		 A C T A G T T G A T C A
BglII  (2008)
1 site		 A G A T C T T C T A G A
SrfI  (2020)
1 site		 G C C C G G G C C G G G C C C G
BamHI  (2027)
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  (2033)
1 site		 C A A T T G G T T A A C
Eco53kI  (2042)
1 site		 G A G C T C C T C G A G
SacI  (2044)
1 site		 G A G C T C C T C G A G
BsrGI  (2047)
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  (2055)
1 site		 G G C G C G C C C C G C G C G G
PstI  (2065)
1 site		 C T G C A G G A C G T C
SbfI  (2065)
1 site		 C C T G C A G G G G A C G T C C
SalI  (2067)
1 site		 G T C G A C C A G C T G
Acc65I  (2073)
1 site		 G G T A C C C C A T G G
AgeI  (2076)
1 site		 A C C G G T T G G C C A
KpnI  (2077)
1 site		 G G T A C C C C A T G G
HindIII  (2085)
1 site		 A A G C T T T T C G A A
NotI  (2092)
1 site		 G C G G C C G C C G C C G G C G
BstZ17I  (2107)
1 site		 G T A T A C C A T A T G
PmlI  (2112)
1 site		 C A C G T G G T G C A C
PaeR7I  (2154)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (2154)
1 site		 C T C G A G G A G C T C
DraIII  (2165)
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.
BbvCI  (2175)
1 site		 C C T C A G C G G A G T C G
Bpu10I  (2175)
1 site		 C C T N A G C G G A N T C G
Cleavage may be enhanced when more than one copy of the Bpu10I recognition sequence is present.
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.
BlpI  (2212)
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.
Bsu36I  (2289)
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  (2318)
1 site		 T T A A T T A A A A T T A A T T
PflFI  (2542)
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  (2542)
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.
KasI  (2812)
1 site		 G G C G C C C C G C G G
NarI  (2813)
1 site		 G G C G C C C C G C G G
* Blocked by Dcm methylation.
Efficient cleavage requires at least two copies of the NarI recognition sequence.
SfoI  (2814)
1 site		 G G C G C C C C G C G G
PluTI  (2816)
1 site		 G G C G C C C C G C G G
Efficient cleavage requires at least two copies of the PluTI recognition sequence.
BsmBI  (2836)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
Esp3I  (2836)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
HpaI  (2949)
1 site		 G T T A A C C A A T T G
lacI
2724 .. 3806  =  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
2724 .. 3806  =  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).
GST
1179 .. 1835  =  657 bp
219 amino acids  =  25.6 kDa
Product: glutathione S-transferase from Schistosoma japonicum
GST
1179 .. 1835  =  657 bp
219 amino acids  =  25.6 kDa
Product: glutathione S-transferase from Schistosoma japonicum
6xHis
1872 .. 1889  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
1872 .. 1889  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
S-Tag
1899 .. 1943  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
S-Tag
1899 .. 1943  =  45 bp
15 amino acids  =  1.7 kDa
Product: affinity and epitope tag derived from pancreatic ribonuclease A
thrombin site
1959 .. 1976  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
thrombin site
1959 .. 1976  =  18 bp
6 amino acids  =  627.7 Da
Product: thrombin recognition and cleavage site
enterokinase site
1995 .. 2009  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
enterokinase site
1995 .. 2009  =  15 bp
5 amino acids  =  606.5 Da
Product: enterokinase recognition and cleavage site
HSV tag
2121 .. 2153  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
HSV tag
2121 .. 2153  =  33 bp
11 amino acids  =  1.2 kDa
Product: HSV (herpes simplex virus) epitope tag
AmpR
4121 .. 4981  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4121 .. 4189  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4121 .. 4981  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   4190 .. 4981  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4121 .. 4981  =  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
5152 .. 5740  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5152 .. 5740  =  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
AmpR promoter
4016 .. 4120  =  105 bp
AmpR promoter
4016 .. 4120  =  105 bp
lacI promoter
3807 .. 3884  =  78 bp
lacI promoter
3807 .. 3884  =  78 bp
T7 terminator
2223 .. 2270  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
T7 terminator
2223 .. 2270  =  48 bp
transcription terminator for bacteriophage T7 RNA polymerase
lac operator
1111 .. 1135  =  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
1111 .. 1135  =  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
1092 .. 1110  =  19 bp
promoter for bacteriophage T7 RNA polymerase
T7 promoter
1092 .. 1110  =  19 bp
promoter for bacteriophage T7 RNA polymerase
RBS
1166 .. 1171  =  6 bp
ribosome binding site
RBS
1166 .. 1171  =  6 bp
ribosome binding site
MCS
2008 .. 2115  =  108 bp
multiple cloning site
MCS
2008 .. 2115  =  108 bp
multiple cloning site
ORF:  1840 .. 2169  =  330 bp
ORF:  109 amino acids  =  12.2 kDa
ORF:  3982 .. 4224  =  243 bp
ORF:  80 amino acids  =  9.3 kDa
ORF:  2516 .. 2941  =  426 bp
ORF:  141 amino acids  =  15.5 kDa
ORF:  4121 .. 4981  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1179 .. 2165  =  987 bp
ORF:  328 amino acids  =  37.0 kDa
ORF:  2704 .. 2967  =  264 bp
ORF:  87 amino acids  =  8.9 kDa
ORF:  3556 .. 3921  =  366 bp
ORF:  121 amino acids  =  13.1 kDa
ORF:  4585 .. 4851  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2724 .. 3683  =  960 bp
ORF:  319 amino acids  =  34.1 kDa
ORF:  452 .. 1018  =  567 bp
ORF:  188 amino acids  =  21.4 kDa
ORF:  1619 .. 1867  =  249 bp
ORF:  82 amino acids  =  9.1 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

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
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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