Shop All Bacterial Expression Vectors

pTrcHis A, B, & C Bacterial Expression Vectors (Invitrogen™)

Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli. These vectors feature:

• High-level regulated transcription from the trc promoter
• Enhanced translation efficiency of eukaryotic genes in E. coli
• The lacO operator and lacIq repressor gene for transcriptional regulation in any E. coli strain

This particular vector offers:

• N-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an anti-HisG antibody
• N-terminal Xpress™ epitope for easy detection with an anti-Xpress™ antibody
• Enterokinase cleavage site for removal of fusion tag

For C-terminal polyhistidine tag and c-myc epitope, please see our pTrcHis2 A, B, & C Vector.

aLICator LIC Cloning and Expression Kit 4 (N-terminal His-tag/WQ) (Thermo Scientific™)

Thermo Scientific aLICator™ LIC Cloning and Expression System is designed for fast and efficient ligation independent cloning and tight regulation of gene expression in E. coli. The pLATE bacterial expression vectors are designed for high levels of target protein expression in concert with minimal background (uninduced) expression, which permits expression of proteins that are toxic to E. coli cells. To streamline and facilitate the process of insert cloning into the expression vector, the aLICator system uses directional LIC cloning technology, a rapid procedure that provides high-cloning efficiencies.

The tightly regulated expression and fast, efficient directional cloning makes the aLICator LIC Cloning and Expression System the best choice for routine and toxic gene cloning and expression in E. coli.

Highlights

• High efficiency LIC cloning
• Tight control of gene expression
• High yield expression
• Versatile—tagged or untaged protein expression with tag removal option

Applications

• Directional PCR product cloning
• Tightly regulated protein expression
• Expression of toxic genes

Principle

The aLICator LIC cloning system uses directional LIC cloning technology to streamline and facilitate cloning into an expression vector. LIC ensures high-cloning efficiencies of more than 95% and eliminates the need for ligation and restriction enzyme digestion steps.

The LIC method uses T4 DNA polymerase to create specific 14 to 21 nucleotide single-stranded overhangs on the pLATE vectors and DNA inserts. T4 DNA polymerase has two enzymatic activities: 5'→3' polymerase activity and 3'→5' exonuclease activity. The exonuclease activity removes nucleotides from the 3' ends of the DNA while the polymerase activity restores the chain using dNTPs and the complementary DNA strand as a template. In the LIC protocol, only dGTP is included in the reaction, causing the 3'→5'-exonuclease and 5'→3'-polymerase activities to equilibrate at the first occurrence of cytosine in the complementary strand. After annealing, the LIC vector and insert are transformed into competent E. coli cells without the use of T4 DNA ligase. Covalent bond formation at the vector-insert junctions occurs within the cell to yield circular plasmid.

Features

The system consists of four kits based on the pLATE series of bacterial expression vectors:

aLICator™ LIC Cloning and Expression Kit 1 - pLATE11 vector, untagged protein expression.
aLICator™ LIC Cloning and Expression Kit 2 (N-terminal His-tag/EK)—pLATE51 vector, N-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Kit 3 (C-terminal His-tag)—pLATE31 vector, C-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Kit 4 (N-terminal His-tag/WQ)—pLATE 52 vector, N-terminal His-tag protein expression, WELQut cleavage.
aLICator™ LIC Cloning and Expression Set 1 (All-in-One/EK)—pLATE11, pLATE51 and pLATE31 vectors, choice of untagged, N- or C-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Set 2 (All-in-One/WQ)—pLATE11, pLATE52, and pLATE31 vectors, choice of untagged, N- or C-terminal His tag protein expression, WELQut cleavage.

For proteins with a known preference for either the N- or C-terminal 6xHis-tag position, using the appropriate N- or C-terminal kit is recommended. When the protein structure and features are not well known, it is recommended to clone into all three vectors and determine the most compatible vector for further research.

Related Products
aLICator LIC Cloning and Expression Set 2 (All-in-One/WQ)
CloneJET PCR Cloning Kit
Fast DNA End Repair Kit
aLICator LIC Cloning and Expression Kit 1 (untagged)

pTrcHis2 A, B, & C Bacterial Expression Vectors (Invitrogen™)

Our pTrcHis A, B, & C vectors are designed to offer enhanced translation initiation and high-level expression in E. coli. These vectors feature:

• High-level regulated transcription from the trc promoter
• Enhanced translation efficiency of eukaryotic genes in E. coli
• The lacO operator and lacIq repressor gene for transcriptional regulation in any E. coli strain

This particular vector offers:

• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an anti-His(C-term) antibody
• C-terminal c-myc epitope for easy detection of fusion proteins with an anti-myc antibody

For N-terminal polyhistidine tag and Xpress™ epitope, please see our pTrcHis A, B, & C Vector.

Gateway™ pDEST™17 Vector (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately:
Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

Gateway™ pET-DEST42 Vector (Invitrogen™)

The Champion™ pET-DEST42 Gateway® destination vector is designed for rapid cloning with a Gateway® entry clone using lambda phage site-specific recombination and subsequent high-level prokaryotic expression controlled by the strong bacteriophage T7 promoter. Expression is induced by the production of T7 RNA polymerase in BL21(DE3) E. coli. The Champion™ pET-DEST42 destination vector offers:

• Bacteriophage T7lac promoter for high-level expression
O operator sequences for lac repressor binding to ensure tighter regulation of transcription
• pBR322 ori for minimal basal expression
• C-terminal V5 and 6xHis tags for efficient detection and purification
R sites for efficient recombination with any attL-flanked Gateway® entry vector

aLICator LIC Cloning and Expression Set 2 (All-in-One/WQ) (Thermo Scientific™)

Thermo Scientific aLICator™ LIC Cloning and Expression System is designed for fast and efficient ligation independent cloning and tight regulation of gene expression in E. coli. The pLATE bacterial expression vectors are designed for high levels of target protein expression in concert with minimal background (uninduced) expression, which permits expression of proteins that are toxic to E. coli cells. To streamline and facilitate the process of insert cloning into the expression vector, the aLICator system uses directional LIC cloning technology, a rapid procedure that provides high-cloning efficiencies.

The tightly regulated expression and fast, efficient directional cloning makes the aLICator LIC Cloning and Expression System the best choice for routine and toxic gene cloning and expression in E. coli.

Highlights

• High efficiency LIC cloning
• Tight control of gene expression
• High yield expression
• Versatile—tagged or untaged protein expression with tag removal option

Applications

• Directional PCR product cloning
• Tightly regulated protein expression
• Expression of toxic genes

Principle

The aLICator LIC cloning system uses directional LIC cloning technology to streamline and facilitate cloning into an expression vector. LIC ensures high-cloning efficiencies of more than 95% and eliminates the need for ligation and restriction enzyme digestion steps.

The LIC method uses T4 DNA polymerase to create specific 14 to 21 nucleotide single-stranded overhangs on the pLATE vectors and DNA inserts. T4 DNA polymerase has two enzymatic activities: 5'→3' polymerase activity and 3'→5' exonuclease activity. The exonuclease activity removes nucleotides from the 3' ends of the DNA while the polymerase activity restores the chain using dNTPs and the complementary DNA strand as a template. In the LIC protocol, only dGTP is included in the reaction, causing the 3'→5'-exonuclease and 5'→3'-polymerase activities to equilibrate at the first occurrence of cytosine in the complementary strand. After annealing, the LIC vector and insert are transformed into competent E. coli cells without the use of T4 DNA ligase. Covalent bond formation at the vector-insert junctions occurs within the cell to yield circular plasmid.

Features

The system consists of four kits based on the pLATE series of bacterial expression vectors:

aLICator™ LIC Cloning and Expression Kit 1 - pLATE11 vector, untagged protein expression.
aLICator™ LIC Cloning and Expression Kit 2 (N-terminal His-tag/EK)—pLATE51 vector, N-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Kit 3 (C-terminal His-tag)—pLATE31 vector, C-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Kit 4 (N-terminal His-tag/WQ)—pLATE 52 vector, N-terminal His-tag protein expression, WELQut cleavage.
aLICator™ LIC Cloning and Expression Set 1 (All-in-One/EK)—pLATE11, pLATE51 and pLATE31 vectors, choice of untagged, N- or C-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Set 2 (All-in-One/WQ)—pLATE11, pLATE52, and pLATE31 vectors, choice of untagged, N- or C-terminal His tag protein expression, WELQut cleavage.

For proteins with a known preference for either the N- or C-terminal 6xHis-tag position, using the appropriate N- or C-terminal kit is recommended. When the protein structure and features are not well known, it is recommended to clone into all three vectors and determine the most compatible vector for further research.

Related Products
aLICator LIC Cloning and Expression Set 1 (All-in-One/EK)
aLICator LIC Cloning and Expression Kit 4 (N-terminal His-tag/WQ)
aLICator LIC Cloning and Expression Kit 3 (C-terminal His-tag)
aLICator LIC Cloning and Expression Kit 2 (N-terminal His-tag/EK)
aLICator LIC Cloning and Expression Kit 1 (untagged)

pBAD/His Kit (Invitrogen™)

The pBAD/His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The vector pBAD/His allows you to express your protein with an N-terminal tag. The vector provides:

• The araBAD promoter for tightly regulated expression
• Translation initiation signals optimized for E. coliexpression
• N-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin and detection with an Anti-HisG Antibody
• N-terminal Xpress™ epitope for detection and analysis with an Anti-Xpress™ Antibody
• Enterokinase cleavage site for removing the N-terminal tag following purification

Three vectors are provided (A, B, and C). Each has the N-terminal tag in a different reading
frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

pBAD/gIII Kit (Invitrogen™)

The pBAD/gIII Kit provides all of the necessary reagents to express recombinant proteins under the tight regulation of the araBAD promoter. To simplify purification, the vector pBAD/gIII encodes a leader peptide which directs the recombinant protein into the
periplasmic space. The vector provides:

• The araBAD promoter for tightly regulated expression
• Translation initiation signals for optimized E. coliexpression
• Leader peptide from the bacteriophage fd gene III protein (gIII) for secreted expression
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin
• C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody

A set of three vectors is provided (A, B, and C). Each has the C-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

Gateway™ pDEST™24 Vector (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

pRSET A, B, & C Bacterial Expression Vectors (Invitrogen™)

The pRSET vector is designed for high-level prokaryotic expression controlled by the strong bacteriophage T7 promoter. Expression is induced by the production of T7 RNA polymerase in BL21(DE3) E. coli. These cells also produce T7 lysozyme to reduce basal expression of target genes. The pRSET vector offers:

• Bacteriophage T7 promoter for high-level expression
• T7 gene 10 sequence to provide protein stability
• N-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-HisG Antibody
• N-terminal Xpress™ epitope for detection with the Anti-Xpress™ Antibody
• Enterokinase cleavage site for removal of fusion tag

A set of three vectors is provided (A, B, and C). Each has the N-terminal tag coding sequence in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

aLICator LIC Cloning and Expression Set 1 (All-in-One/EK) (Thermo Scientific™)

Thermo Scientific aLICator™ LIC Cloning and Expression System is designed for fast and efficient ligation independent cloning and tight regulation of gene expression in E. coli. The pLATE bacterial expression vectors are designed for high levels of target protein expression in concert with minimal background (uninduced) expression, which permits expression of proteins that are toxic to E. coli cells. To streamline and facilitate the process of insert cloning into the expression vector, the aLICator system uses directional LIC cloning technology, a rapid procedure that provides high-cloning efficiencies.

The tightly regulated expression and fast, efficient directional cloning makes the aLICator LIC Cloning and Expression System the best choice for routine and toxic gene cloning and expression in E. coli.

Highlights

• High efficiency LIC cloning
• Tight control of gene expression
• High yield expression
• Versatile—tagged or untaged protein expression with tag removal option

Applications

• Directional PCR product cloning
• Tightly regulated protein expression
• Expression of toxic genes

Principle

The aLICator LIC cloning system uses directional LIC cloning technology to streamline and facilitate cloning into an expression vector. LIC ensures high-cloning efficiencies of more than 95% and eliminates the need for ligation and restriction enzyme digestion steps.

The LIC method uses T4 DNA polymerase to create specific 14 to 21 nucleotide single-stranded overhangs on the pLATE vectors and DNA inserts. T4 DNA polymerase has two enzymatic activities: 5'→3' polymerase activity and 3'→5' exonuclease activity. The exonuclease activity removes nucleotides from the 3' ends of the DNA while the polymerase activity restores the chain using dNTPs and the complementary DNA strand as a template. In the LIC protocol, only dGTP is included in the reaction, causing the 3'→5'-exonuclease and 5'→3'-polymerase activities to equilibrate at the first occurrence of cytosine in the complementary strand. After annealing, the LIC vector and insert are transformed into competent E. coli cells without the use of T4 DNA ligase. Covalent bond formation at the vector-insert junctions occurs within the cell to yield circular plasmid.

Features

The system consists of four kits based on the pLATE series of bacterial expression vectors:

aLICator™ LIC Cloning and Expression Kit 1 - pLATE11 vector, untagged protein expression.
aLICator™ LIC Cloning and Expression Kit 2 (N-terminal His-tag/EK)—pLATE51 vector, N-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Kit 3 (C-terminal His-tag)—pLATE31 vector, C-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Kit 4 (N-terminal His-tag/WQ)—pLATE 52 vector, N-terminal His-tag protein expression, WELQut cleavage.
aLICator™ LIC Cloning and Expression Set 1 (All-in-One/EK)—pLATE11, pLATE51 and pLATE31 vectors, choice of untagged, N- or C-terminal His-tag protein expression.
aLICator™ LIC Cloning and Expression Set 2 (All-in-One/WQ)—pLATE11, pLATE52, and pLATE31 vectors, choice of untagged, N- or C-terminal His tag protein expression, WELQut cleavage.

For proteins with a known preference for either the N- or C-terminal 6xHis-tag position, using the appropriate N- or C-terminal kit is recommended. When the protein structure and features are not well known, it is recommended to clone into all three vectors and determine the most compatible vector for further research.

Related Products
aLICator LIC Cloning and Expression Kit 4 (N-terminal His-tag/WQ)
aLICator LIC Cloning and Expression Kit 3 (C-terminal His-tag)
aLICator LIC Cloning and Expression Set 2 (All-in-One/WQ)
aLICator LIC Cloning and Expression Kit 1 (untagged)
aLICator LIC Cloning and Expression Kit 2 (N-terminal His-tag/EK)

pBAD/Myc-His Kit (Invitrogen™)

The pBAD/Myc-His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The pBAD/Myc-His vector expresses native proteins or fusion proteins with a C-terminal tag. The vector provides:

• The araBAD promoter for tightly regulated expression
• Translation initiation signals optimized for E. coliexpression
• C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin or detection with an Anti-His(C-term) Antibody
• C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody

Three vectors are provided (A, B, and C). Each has the C-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.

Gateway™ pDEST™14 Vector (Invitrogen™)

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

pBAD202 Directional TOPO™ Expression Kit (Invitrogen™)

Using restriction enzymes to clone your gene into an expression vector often forces you to compromise the final sequence of your insert (Figure 1A), especially when there are no useful restriction sites close to your genes coding sequence. This may result in suboptimal spacing of expression elements or incorporation of non-native amino acid residues, which can reduce your expression levels and/or cause the production of non-functional protein.

In addition to being a more effective way to clone, TOPO® Cloning eliminates these potential expression problems. TOPO® Expression Vectors enable you to insert the exact DNA sequence you require simply by performing PCR with appropriately designed primers. Your PCR product is cloned at a high efficiency in only five minutes into a topoisomerase I-activated expression vector. The resulting recombinant expression vector contains your exact DNA sequence without any non-coding regions (Figure 1B).

Many of Invitrogen's powerful expression vectors are available adapted for one-step TOPO® cloning and expression of PCR products. In addition, several expression vectors are now adapted for Directional TOPO® Cloning, enabling you to use proofreading polymerases and to clone your PCR products in a specific orientation.

Champion™ pET302/NT-His and pET303/CT-His Vector Kit (Invitrogen™)

The Champion™ pET302/NT-His and pET303/CT-His Vector Kit is designed for cloning a gene of interest via restriction enzyme(s) and ligase (REaL) and subsequent high-level expression from the strong bacteriophage T7 promoter. In addition to the T7 promoter, each vector contains only the necessary functional elements and an N- or C-terminal 6xHis tag (pET302/NT-His and pET303/CT-His, respectively) for convenient purification and detection (Figure 1). Expression levels obtained from these vectors may be higher than those obtained from another suppliers pET vector (Figure 2). To maximize expression, use with MagicMedia™ E. coli Expression Medium.


Contents and Storage:

The Champion™ pET302/NT-His and pET303/CT-His Vector Kit includes 6 µg each of pET302/NT-His and pET303/CT-His and 10 µg of a control vector. Store at -20“C. All components are guaranteed stable for 6 months when properly stored.