pOptiVEC-TOPO circularized vector哺乳动物细胞高效表达载体DHFR筛选，环状质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心

pOptiVEC-TOPO circularized vector哺乳动物细胞高效表达载体DHFR筛选，环状质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心
In the pOptiVEC-TOPO vector, the transcription of the gene of interest is separated from the dihydrofolate reductase (DHFR) auxotrophic selection marker by an internal ribosome entry site (IRES), allowing transcription of the gene of interest and the selection marker on the same mRNA.
The pOptiVEC-TOPO vector contains the following elements:
• Human cytomegalovirus (CMV) immediate-early promoter/enhancer for high-level gene expression in a wide range of mammalian cells
• TOPO Cloning site for rapid and efficient cloning of Taq-amplified PCR products
• Internal Ribosome Entry Site (IRES) from the encephalomyocarditis virus (EMCV) for cap-independent translation of DHFR
• Dihydrofolate reductase (DHFR) gene for auxotrophic selection of transfected DG44 cells and for genomic amplification of stable cell lines using methotrexate (MTX)
• The Herpes Simplex Virus thymidine kinase polyadenylation signal for proper termination and processing of the recombinant transcript
• pUC origin for high copy replication and maintenance of the plasmid in E. coli
• Ampicillin (bla) resistance gene for selection in E. coli

To obtain the pOptiVEC-TOPO expression construct containing your gene of interest, your PCR primer design must include:
• A Kozak consensus sequence
• A mammalian secretion signal upstream of your gene of interest (if you wish to produce secreted protein)
• A stop codon at the end of your gene of interest
Secretion signal
If you wish for your protein of interest to be secreted from mammalian DG44 cells, your gene of interest must include a mammalian secretion signal. To direct secretion of your protein of interest, you can include the endogenous secretion signal of your protein of interest, or add one such as the murine Ig κ-chain leader sequence (Coloma et al., 1992) using PCR.
Primer Sequence
CMV forward 5’-CGCAAATGGGCGGTAGGCGTG-3’
EMCV IRES reverse 5’- CCTTATTCCAAGCGGCTTCG-3’

【Supplier来源】BioVector NTCC Inc.
TEL：+86-010-53513060
【Website网址】 http://www.biovector.net