pAAVdual-CAG-CcdB

The pAAVdual-CAG-CcdB plasmid serves as a destination vector for efficient ORF cloning into the pAAVdual system using Gateway technology. This system is part of AAVnerGene’s AAVdual platform, which integrates the AAV vector plasmid (pAAVtri) and the Ad helper plasmid (mini-pHelper) into a single vector. The CAG promoter is a synthetic hybrid promoter widely used in molecular biology to drive high levels of gene expression in mammalian cells. It is composed of three key elements: CMV Immediate-Early Enhancer: Derived from the cytomegalovirus (CMV), this enhancer sequence provides strong and ubiquitous transcriptional activation in a variety of cell types. Chick Beta-Actin Promoter: The core promoter region is derived from the chicken beta-actin gene, which is known for its high expression levels in many cell types, particularly in the context of gene expression in vertebrates. Rabbit Beta-Globin Intron: An intronic sequence from the rabbit beta-globin gene is included to enhance transcriptional efficiency and mRNA stability, further boosting expression levels. It is particularly valued for its strength and broad activity across various cell types, making it a popular choice for applications such as gene therapy, transgenic animal models, and research studies that require robust and consistent gene expression. 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-CAG-CcdB destination plasmid along with the pENTR-GOI vector from the ORFeome collections, researchers can efficiently generate pAAVdual-CAG-GOI plasmids. These plasmids can then be used for AAV packaging within the AAVdual system, enabling the production of corresponding AAV vectors.

• AAV Cassette: AAV-CAG-CcdB
• AAV Cassette Size: 3,461 bp
• Plasmid Size: 11,935 bp
• Plasmid Backbone: pAAVdual
• AAV Genome Type: ssAAV
• ITRs:  L-ITR (119 bp) + R-ITR (130 bp)
• Promoter: CAG
• Transgene: CcdB
• Bacterial Resistance(s) : Kana
• Storage:  -20°C for long term (> 1 year)
• Shipping: Dry ice.
• Notice to Purchaser:  MTA is required. All mini-pHelper based products, including mini-pHelper itself, pAAVdual and pAAVone plasmids,  are sold under license of US 20240229067. Rights to use these products are limited to non-commercial research only. No other rights are conveyed. Inquiry into the availability of a license to broader rights or the use of these products for commercial purposes should be directed to AAVnerGene Inc, 9620 Medical Center Dr, Suite 100, Rockville, MD, 20850. Purchase of these products does not grant rights to : (1) offer the vectors or any derivatives thereof for resale; or (2) to distribute or transfer the vector or any derivative thereof to third parties.
• Cloning Services: AAVnerGene provides cloning services based on this plasmid backbone with $300/clone.
• AAV Package Services: AAVnerGene provides affordable AAV production services at various scales(from 1e12 to 1e16 vg), in both suspension and adherent cultured cells (HEK 293T or HEK 293), with flexible purification methods (CsCl, Iodixanol, or AAV-X resin).
• AAV Analytical Services: We also offer advanced AAV characterization services,  including AAV purity, vector genome identity, ITR integrity, DNA impurities, empty/full ratios, and a variety of comprehensive analyses based on custom requests.