New vectors for the yeast genome editing using CRISPR/Cas9 were constructed. A system for cloning of new targets using the standard methods (PCR‒restriction‒ligation) was developed and successfully applied. The constructed vectors allowed us to obtain the sup35-25 mutants, deletion of the PSH1 gene and disruption of the NAM7 (UPF1). A convenient method for identifying plasmids with a new target was tested. A detailed description of the cloning technique used and selection of plasmids with the new targets is provided.