Traditional bacterial fermentation techniques used to manufacture plasmid are time-consuming, expensive, and inherently unstable. The production of sufficient GMP grade material thus imposes a major bottleneck on industrial scale manufacturing of lentiviral vectors (LVV). Touchlight’s linear doggybone DNA (dbDNA™) is an enzymatically amplified DNA vector produced with exceptional speed through an in vitro dual enzyme process, enabling industrial scale manufacturing of GMP material in a fraction of the time required for plasmid. Here, we demonstrate that dbDNA can be optimised for the manufacture of high titre LVV.