pcDNA™3.4 TOPO™ TA Cloning Kit
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Invitrogen™

pcDNA™3.4 TOPO™ TA Cloning Kit

The pcDNA™ 3.4 TOPO™ vector is a constitutive mammalian expression vector designed to deliver exceptionally high levels of transgene expression.Read more
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Catalog number A14697
Price (USD)
1,628.00
Each
Add to cart
Price (USD)
1,628.00
Each
Add to cart
The pcDNA™ 3.4 TOPO™ vector is a constitutive mammalian expression vector designed to deliver exceptionally high levels of transgene expression. It can be used in suspension-adapted cells, such as Expi293F™, FreeStyle™ 293-F, and FreeStyle™ CHO-S cells, for transient protein expression. The vector can also be used as a Geneticin™-selectable expression plasmid for engineering stable cell lines. The enhanced, full-length CMV promoter and other expression elements typically enable higher expression levels than pcDNA™ 3.1-, 3.2-, and 3.3-based expression constructs.

The pcDNA 3.4 TOPO™ vector contains the following elements:

• Full length human cytomegalovirus (CMV) immediate-early promoter/enhancer for high level gene expression in a wide range of mammalian cells
• WPRE (woodchuck posttranscriptional regulatory element) downstream of the cloning site to enhance transcript expression
• TOPO™ cloning site for rapid and efficient (>85%) cloning of Taq-amplified PCR products
• Neomycin resistance gene for selection of stable cell lines with Geneticin™
• pUC origin for high copy replication and maintenance of the plasmid in E. coli

The pcDNA3.4™ TOPO™ TA Cloning Kit includes TOPO™-adapted plasmid vector, reagents for PCR cloning of your gene of choice into the vector, a sequencing primer for confirmation of insert orientation, and One Shot™ TOP10 competent cells.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Antibiotic Resistance BacterialAmpicillin (AmpR)
Cloning MethodTOPO™-TA
Constitutive or Inducible SystemConstitutive
Delivery TypeTransfection
For Use With (Application)Protein Production, Constitutive Expression
PlasmidHigh Copy Plasmid
Product LineTOPO™
Product TypeTA Cloning Kit
PromoterCMV
Protein TagUntagged
Quantity20 μg
Replication OriginpUC Origin
Selection Agent (Eukaryotic)Geneticin™ (G-418)
Selection Marker EukaryoticNeoR
Selection Marker PromoterSV40 Promoter
VectorpcDNA
Unit SizeEach
Contents & Storage
Store pcDNA™3.4-TOPO™ vector and all cloning reagents at -20°C.
Store One Shot™ TOP10 Chemically Competent E. coli cells at -80°C.

Frequently asked questions (FAQs)

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

Can I use a different 293 cell line with the Expi293 Expression System?

Other 293 cell lines may be used with the Expi293 Expression System. However, before these cell lines may be used for transfection studies, they must be adapted to serum-free, suspension culture in Expi293 Expression Medium and evaluated for transfection and expression.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

What is the difference between pcDNA 3.4-TOPO TA and pcDNA 3.3-TOPO TA vectors?

pcDNA 3.4-TOPO TA vector is an improvement over pcDNA 3.3-TOPO TA vector. It contains the WPRE (Woodchuck Posttranscriptional Regulatory Element) that allows for 2- to 3-fold higher levels of expression than pcDNA 3.3-TOPO TA vector.

Which vector do you recommend using with the Expi293 Expression System?

We recommend using the pcDNA 3.4-TOPO TA vector (Cat. No. A14697). This vector contains the native, full-length CMV promoter and a WPRE (Woodchuck Posttranscriptional Regulatory Element) downstream of the cloning site, both of which contribute to high level gene expression (about 2-3 fold higher expression than with pcDNA3.3 TOPO vector, which in turn provides 2-5 fold higher expression than with a standard pcDNA vector). Of course, the expression level is also protein-dependent.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

View all pcDNA™3.4 TOPO™ TA Cloning Kit FAQs