pAAVdual-EF1a-CcdB

The pAAVdual-EF1a-CcdB plasmid serves as a destination vector for efficient ORF cloning into the pAAVdual system using Gateway technology. This system is part of AAVdual platform, which integrates the AAV vector plasmid (pAAVtri) and the Ad helper plasmid (mini-pHelper) into a single vector. The EF1a promoter, also known as the cytomegalovirus promoter, is a widely used promoter sequence in molecular biology and biotechnology. It is derived from the immediate early promoter of the cytomegalovirus and is commonly used to drive the expression of genes in a variety of experimental and practical applications. The CcdB gene in the plasmid encodes a toxic protein that disrupts DNA gyrase, a bacterial enzyme essential for DNA replication, leading to cell death. This ensures that only bacterial cells with successful recombination events—those replacing the CcdB gene—survive, facilitating positive selection during cloning. By using the pAAVdual-EF1a-CcdB destination plasmid along with the pENTR-GOI vector from the ORFeome collections, researchers can efficiently generate pAAVdual-EF1a-GOI plasmids. These plasmids can then be used for AAV packaging within the AAVdual system, enabling the production of corresponding AAV vectors.

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Background

The ORFeome collections provide a comprehensive set of open reading frames (ORFs) from a genome, with each entry vector carrying an ORF flanked by attL1 and attL2 sites. In Gateway cloning, the entry clone undergoes an LR reaction with the destination vector, where the attL and attR sites recombine. This process transfers the gene of interest from the entry vector into the destination vector, replacing the CcdB gene with the ORF, enabling successful cloning for AAV production.NameSKUPriceBuySelect all