Schizosaccharomyces japonicus is an organism that possesses very unique characteristics especially dimorphism which is the ability to change between yeast and hyphae forms. In this work, our aim was to create a deletion variant of the S. japonicus cgs1 gene that encodes the regulatory subunit of the protein kinase A (pka1) which is involved in the regulation of many hundred genes such as genes of morphological switching and hyphal growth. Deletion of the gene from the chromosome will allow us to investigate the effects of the absence of this gene in the S.japonicus cell as well as its functions. To create a deletion variant of the S. japonicus cgs1 gene, there was isolating the genomic DNA from S. japonicus cells followed by the amplification of the cgs1 gene from its chromosome. The coding region of cgs1 was removed by restriction digestion and the KanMX6 cassette was inserted between the cgs1 flanking sequences. The pJET1.2 vector +cgs1 5’flanking region+KanMX6 cassette+cgs1 3’flanking region was transformed into bacterial cells and five colonies were placed into deep-freezing preservation after colony PCR checking.