Shop All Mammalian Expression Vectors

Jump-In™ Fast Gateway™ System (Invitrogen™)

The Jump-In™ Fast Gateway® System is intended for rapid engineering of mammalian cell lines through the stable and irreversible integration of your gene of interest (GOI) at specific genomic loci (pseudo attP sites). Following stable integration, only 5-10 clones need to be analyzed to identify clones with high expression levels.
The Jump-In™ Fast Gateway® System consists of:

• The Jump-In™ Fast Gateway® Core Kit containing the promoter-less pJTI™ Fast DEST expression vector and a vector for transient expression of the PhiC31 integrase.
• The MultiSite Gateway® Pro Plus kit for cloning of up to 4 gene fragments into the promoter-less pJTI™ Fast DEST expression vector.

pcDNA™3.2/GW/D-TOPO™ Expression Kit (Invitrogen™)

The pcDNA™ vectors are designed for high-level, constitutive expression in a variety of mammalian cell lines. The pcDNA3.2/GW/D-TOPO vector offers the following key features:

•Cytomegalovirus (CMV) promoter for high-level expression
•Adapted for Directional TOPO® Cloning, enabling you to use proofreading polymerases and to clone your PCR products in a specific orientation
•Neomycin resistance gene for stable selection
•C-terminal V5 tag for easy detection
•Ampicillin resistance gene and pUC origin for selection and maintenance in E. coli

TOPO® Cloning
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 our 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.

Jump-In™ Fast Gateway™ Core Kit (Invitrogen™)

The Jump-In™ Fast Gateway® Core Kit is intended for rapid engineering of mammalian cell lines through the stable and irreversible integration of your gene of interest (GOI) at specific genomic loci (pseudo attP sites). Following stable integration, only 5-10 clones need to be analyzed to identify clones with high expression levels.
The Jump-In™ Fast Gateway® Core Kit contains the promoter-less pJTI™ Fast DEST expression vector and a vector for transient expression of the PhiC31 integrase.
Please note: the pJTI™ Fast DEST expression vector requires the cloning of a promoter upstream of the gene of interest, which is facilitated by any of the MultiSite Gateway® Pro Kits.
This kit is also a component of the Jump-In™ Fast Gateway® System.

pEF6/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

T-REx™ Core Kit, with pcDNA™4/TO/myc-His© Vector (Invitrogen™)

A Tetracycline-Regulated Expression System without Viral Transactivators
The T-REx™ System yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).

Specific Activation
The T-REx™ System uses a repressor mechanism that blocks transcription from the powerful CMV promoter in the absence of tetracycline. Because the T-REx™ System elements do not use viral transactivators, you can achieve high-level expression from the complete CMV promoter without secondary, non-specific activation of host genes.

The T-REx™ Mechanism
The T-REx™ transcriptional control elements are illustrated in Figure 1. Two tetracycline operator sequences (TetO2) have been inserted between the TATA box of the CMV promoter and the transcriptional start site. The TetO2 sequence itself has no effect on expression. When the tetracycline repressor protein (TR) is present, it effectively binds the TetO2 sites and blocks transcription initiation. Tetracycline added to the culture medium binds to, and changes the conformation of, the TR protein. This change causes the TR protein to release the TetO2 sites, derepressing transcription from the CMV promoter. The result is high-level expression of the gene of interest (Figure 2). Expression levels can be modulated based on the tetracycline concentration and can be induced to levels that are achieved with constitutive CMV expression vectors.
T-REx™ is a powerful inducible mammalian expression system that allows you to regulate expression from the complete human cytomegalovirus (CMV) enhancer-promoter. T-REx™inducible expression vectors offer the following features:

• Complete CMV enhancer-promoter sequence containing two copies of the tetracycline operator TetO2 sequence for high-level regulated expression
• Zeocin™ or hygromycin resistance gene for effective selection of stable mammalian cell lines
• Large multiple cloning site to simplify cloning

In addition, pcDNA™4/TO/myc-His offers a c-myc epitope for rapid detection of the recombinant protein with an Anti-myc Antibody and a polyhistidine (6xHis) sequence for simple purification of the recombinant protein with nickel-chelating resin and detection with Anti- His(C-term) Antibody.

The regulatory vector, pcDNA™6/TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.

pDisplay™ Mammalian Expression Vector (Invitrogen™)

pDisplay™ is a mammalian expression vector designed to target recombinant proteins to the surface of mammalian cells. Displayed proteins can be analyzed for their ability to interact with known or putative ligands. Proteins of interest are targeted and anchored to the cell surface by cloning the gene of interest in frame with the vectors unique N-terminal secretion signal and the C-terminal transmembrane anchoring domain of platelet-derived growth factor receptor (PDGFR). In contrast to phage display vectors which operate exclusively in prokaryotic cells, the pDisplay™ vector offers the advantage of having the displayed protein of interest processed in mammalian cells. Therefore, recombinant proteins of eukaryotic origin that are expressed from pDisplay™ more closely resemble their native form. This may favor more accurate ligand binding interactions. In addition to the N-terminal cell surface targeting signal and C-terminal transmembrane anchoring domain, the pDisplay™ vector contains the following key features:

• T7 promoter/priming site for in vitro transcription of sense RNA and for sequencing of inserts
• Neomycin resistance marker for stable selection in mammalian cells using Geneticin®
• SV40 origin for replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1 and COS-7)
• Ampicillin resistance gene for selection in E. coli

pJTI™ R4 Exp CMV EmGFP pA Vector

The pJTI™ R4 Exp CMV EmGFP pA vector is a positive control vector for assessing the success of a retargeting reaction with a Jump-In™ parental cell line. When co-transfected with the integrase vector (pJTI™ R4 Int vector) included in the Jump-In™ parental kits, the EmGFP will be expressed and positive cells will fluoresce green.

Ensure the Success of Your Jump-In™ Retargeting Reactions
Successful retargeting of Jump-In™ parental cell lines like the the Jump-In™ GripTite™ HEK293 kit (A14150) is dependent on a variety of factors such as:

Transfection efficiency

• Cell confluency
• Antibiotic selection conditions
• Quality and concentration of DNA
• Retargeting vector to integrase vector ratio

We strongly recommend including a retargeting reaction with the pJTI™ R4 Exp CMV EmGFP pA vector in your Jump-In™ experiment along with negative controls (no plasmid DNA, no integrase vector) so you can easily visualize the results and optimize the retargeting conditions.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

pUB6/V5-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pcDNA™6.2/GW/D-TOPO™ Expression Kit (Invitrogen™)

The pcDNA™ vectors are designed for high-level, constitutive expression in a variety of mammalian cell lines. The pcDNA6.2/GW/D-TOPO vector offers the following key features:

•Cytomegalovirus (CMV) promoter for high-level expression
•Adapted for Directional TOPO® Cloning, enabling you to use proofreading polymerases and to clone your PCR products in a specific orientation
•Blasticidin resistance gene for efficient stable selection
•C-terminal V5 tag for easy detection
•Ampicillin resistance gene and pUC origin for selection and maintenance in E. coli

TOPO® Cloning
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 our 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.

ViraPower™ HiPerform™ T-REx™ Gateway™ Expression System (Invitrogen™)

The ViraPower™ HiPerform™ T-REx™ Gateway® Expression System includes all the components needed to generate lentivirus, including vector kit, 293FT cell line, and the support kit. This kit combines Invitrogen’s ViraPower™ HiPerform™ Lentiviral, T-REx™ and Gateway® technologies to facilitate easy recombination-based cloning and lentiviral-based, regulated (Tetracycline-inducible), high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti6.3⁄ TO⁄ V5-DEST vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements.

Advantages
• Generates replication-incompetent lentivirus for transducing dividing and non-dividing mammalian cells
• Easy recombination-based cloning using Gateway® technology
• Stable, long-term, tetracycline-regulated expression
• Enhanced protein expression, up to 4-fold or greater, compared to traditional lentiviral expression systems

Key Features
• WPRE (Woodchuck Posttranscriptional Regulatory Element) from the woodchuck hepatitis virus, increases transgene expression and cPPT (central Polypurine Tract) from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome, thus increasing viral titer. WPRE and cPPT together produce at least a four-fold increase in protein expression in most cell types, compared to other vectors that do not contain these elements.
• Hybrid promoter consisting of the human cytomegalovirus (CMV) promoter and two tetracycline operator 2 (TetO2) sites for high-level, regulated expression of the target gene
• Blasticidin selection marker for stable selection under control of SV40 promoter

Kit Includes
• ViraPower™ HiPerform™ T-Rex™ Gateway® Vector Kit (Cat # A11144)
• ViraPower™ Bsd Lentiviral Support Kit (Cat # K497000)
• 293FT Cell Line (Cat # R70007)
• Gateway® LR Clonase® II Plus Enzyme Mix (Cat # 12538120)
• One Shot® Stbl3™ Chemically Competent E. coli (Cat # C737303)
• Geneticin® (Cat # 10131035)
• pENTR™ Gus positive control plasmid

For research use only. Not intended for any therapeutic or diagnostic use.

pCMV-Red Firefly Luc Vector for Luciferase Assays (Thermo Scientific™)

Thermo Scientific pCMV-Red Firefly Luc is a constitutive expression vector having the luciferase gene under the CMV (Cytomegalovirus) promoter for co-transfection and normalization control to account for experimental variation in combination with other reporters in a gene regulation study using the intracellular red firefly luciferase reporter with excellent light intensity..

Features of Red Firefly Luc:
• Red Firefly Luc Vectors contains a mutant form of the firefly luciferase gene that has a red-shifted emission spectrum.
• pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential.
• pCMV and pTK vectors have the luciferase gene under the CMV promoter and Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter, respectively.
• These pCMV and pTK constitutive expression vectors can be used as normalization controls to account for experimental variation in combination with other reporters.

These vectors are subject to a limited use label license.

More Product Data
Highly sensitive multiplex luciferase reporter assays
Monitoring neuronal differentiation using multiplexed luciferase reporters
Activation of the antioxidant response pathway by pesticide chemicals

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pMCS-Red Firefly Luc Vector for Luciferase Assays
pTK-Red Firefly Luc Vector for Luciferase Assays

ViraPower™ Zeo Lentiviral Support Kit (Invitrogen™)

The ViraPower™ Zeo Lentiviral Support Kit contains the necessary components to transfect 293FT cells to generate viral particles containing the target gene. The kit contains the ViraPower™ Lentiviral Packaging Mix, Lipofectamine® 2000 Reagent and Zeocin™ and is designed to be used with a ViraPower™ Lentiviral expression vector that has the Zeocin™ marker for stable selection. The kit is compatible with all of our ViraPower™ Lentiviral vectors, including the ViraPower™ HiPerform™ Lentiviral vectors.

Kit includes
• ViraPower™ Lentiviral Packaging Mix
• Lipofectamine® 2000 (Cat # 11668027)
• Zeocin™ solution

For research use only. Not intended for any therapeutic or diagnostic use.

pOptiVEC™-TOPO™ TA Cloning™ Kit (Gibco™)

The OptiCHO™ Express Kit is designed for efficient growth and transfection of dihydrofolate reductase- deficient (DHFR-) Chinese hamster ovary (CHO) DG44 cells in suspension culture. The kit provides a complete workflow protocol as well as the necessary components for efficient transfection of DHFR- DG44 cells in a serum-free environment and subsequent stable cell line development, including:
• pOptiVEC™-TOPO® vector - a bicistronic cloning vector containing a DHFR gene for stable cell line development in CHO-DG44 cells (Figure 1); the vector is TOPO® adapted and enables cloning of a PCR product containing your gene of interest with >85% efficiency
• FreeStyle™ MAX Transfection Reagent - an animal origin-free transfection reagent offering high transfection efficiencies and consistent results
• CD OptiCHO™ Medium - a chemically defined, protein-free medium (without hypoxanthine or thymidine) specifically designed for high-density suspension cell culture of recombinant CHO-DG44 cells and secreted protein yield
• CD DG44 Medium - a chemically defined, protein-free medium supplemented with hypoxanthine and thymidine to support optimal growth of DG44 cells in suspension culture
• DG44 cells - preadapted to CD DG44 medium and selected for superior cell growth and transfection efficiencies

In addition, the OptiCHO™ Express Kit provides GIBCO® OptiPRO™ SFM for improved DNA-lipid complex formation, L-glutamine (provided separately for increased media stability), and Pluronic® F-68 to control shear forces in suspension cultures.

Storage:
Store Pluronic® F-68 at 15°C to 30°C. Store CD DG44 Medium, OptiPRO™ SFM, CD OptiCHO™ Medium, and FreeStyle™ MAX Transfection Reagent at 2°C to 8°C in the dark. Store L-glutamine, pOptiVEC™-TOPO® vector, CMV Forward Sequencing Primer, and EMCV IRES Reverse Sequencing Primer at -5°C to -20°C. Store One Shot® TOP10 Chemically Competent E. coli cells at -80°C. Store DG44 cells in liquid nitrogen.

pEF1/myc-His A, B, & C Mammalian Expression Vectors (Invitrogen™)

All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.

pShooter™ Mammalian Expression Vector (pCMV/myc/cyto) (Invitrogen™)

The pShooter™ vectors are designed to localize recombinant proteins to the nucleus, mitochondria, endoplasmic reticulum, or cytoplasm. In addition to the localization signal, each pShooter™ vector contains:

• Strong mammalian promoter (CMV or EF-1É) for constitutive expression of your protein of interest
• C-terminal c-myc epitope for detection with an Anti-myc Antibody
• Bovine Growth Hormone (BGH) polyadenylation site for mRNA stability
• f1 origin for single-stranded rescue of sense DNA
• Neomycin resistance gene for selection of mammalian cells with Geneticin¤ selection agent
• Ampicillin resistance and pUC origin for selection and maintenance in E. coli

Each vector is provided with a positive control plasmid. The positive control expresses GFP and targets it to the same subcellular location as the pShooter™ vector.