pcDNA3.2/GW/D-TOPO Expression Kit
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Invitrogen
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Product
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pcDNA3.2/GW/D-TOPO Expression Kit
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Company
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Invitrogen
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Price
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Pricing Info
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More Information
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View Company Product Page
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Applications
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Mammalian Expression and Cloning
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Conjugate/Tag/Label
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V5-tagged
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Domain/Region/Terminus
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C terminus
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Host
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Mammalian
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Promoter
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CMV
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Quantity
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20 reactions
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Resistance
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ampicillin/ neomycin
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Type
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Plasmid
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Vector Name
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pcDNA3.2/V5/GW-CAT, pcDNA3.2/V5/GW/D-TOPO
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Catalog Number
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K244020
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Selectable Marker
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Geneticin® (G-418)
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Description
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 , 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.