Name: pAAVone-AAVs-GFAP-CcdB - AAVnerGene
SKU: SE004013
Availability: InStock

pAAVone-AAVs-GFAP-CcdB

The pAAVone-AAVs-GFAP-CcdB plasmids serves as destination vectors for efficient ORF cloning into the AAVpme system using Gateway technology. This system is part of AAVnerGene’s AAVone platform, which integrates the AAV vector plasmid (pAAVtri) and the Ad helper plasmid (mini-pHelper) and AAV heper plasmid (pRCap) into a single plasmid. 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 pAAVone-AAVs-GFAP-CcdB destination plasmids along with the pENTR-GOI vectors from the ORFeome collections, researchers can efficiently generate pAAVone-AAVs-GFAP-GOI plasmids. These plasmids can then be used for AAV packaging with the AAVone system, enabling the production of corresponding AAV vectors.

AAV Cassette:AAV-GFAP-CcdB
Plasmid Backbone: pAAVone
AAV Genome Type: ssAAV
ITRs: L-ITR (119 bp) + R-ITR (130 bp)
AAV Serotypes: selectable
Promoter: GFAP
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.

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The pAAVone-AAVs-Pro-CcdB plasmids serve as destination vectors for the efficient cloning of ORFs into the pAAVone backbone, which can produce regular single strand AAV (ssAAV) viruses using AAVnerGene’s novel AAVone system. These plasmids contain AAV heper genes (Rep and Cap), Ad helper genes (E2A, E4orf6, VA RNA) and AAV genomes carried CcdB gene. The CcdB gene is flanked by attR1 and attR2 recombination sites. The CcdB gene functions as a toxin, ensuring only bacterial cells that successfully undergo recombination and replace the CcdB gene with the desired ORF survive.

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.