FIELD: biotechnology; genetic engineering.
SUBSTANCE: disclosed is a plasmid for making modifications in the genome of bacteria of the genus Bacillus using a non-homologous end connection repair system containing the cpf1 gene under the control of an inducible promoter, a gene coding a repressor for a corresponding inducible promoter, an operator sequence cre for additional repression of the cpfl gene by the CcpA repressor, a region for embedding a sequence guiding a gRNA, comprising a spacer to a DNA target, located under the control of a strong constitutive promoter. Also disclosed is a method for modifying the genome of bacteria of the genus Bacillus along the path of non-homologous connection of ends using said plasmid.
EFFECT: group of inventions provides controlled expression of nuclease Cpf1 in bacteria of the genus Bacillus and enables to introduce targeted modifications into the genome of bacteria of the genus Bacillus without using antibiotic resistance markers.
14 cl, 12 dwg, 2 tbl, 9 ex
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