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. 2018 Oct 9;9:2307. doi: 10.3389/fmicb.2018.02307

FIGURE 1.

FIGURE 1

Schematic maps of the developed method. (A) A schematic of the helper plasmid p46Cpf1, in which λ Red recombinases (Gam, Bet, and Exo) are expressed under the control of Ptet, and Cas12a is expressed under the control of ParaB. (B) A schematic of the donor plasmid pTc-GLP, which encodes crRNAs constitutively and provides the templates for recombination at three loci (galK, lacZ, and pyrF). (C) General outline of the multiplex gene insertion method. It takes 8 days to construct a modified plasmid-free strain.