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We initiated an evolution campaign with seven PANCE passages on GD_M, followed by 12 passages of high-stringency dual selection penalizing proteases that cleave VAMP1, and concluded with 16 passages on a circuit using a weaker promoter in front of the GD_M substrate to increase positive selection stringency (Extended Data Fig

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[DOI] [PubMed] [Google Scholar] 22.Maxipime [package insert] Princeton, New Jersey: Bristol Meyers Squibb Company