Abstract: 【Objective】 To analyze the wild plasmid sequence characteristics of Staphylococcus aureus methicillin-resistant strains and vancomycin-resistant strains in the hope of helping the study of the formation of super-drug-resistant bacteria. 【Methods】 The multidrug-resistant strain D353 was identified by 16s rRNA, and the sequence features, genes, function and replication of the plasmid pD353 were analyzed by BLAST, MEGA 11, Mauve, and trf. 【Results】 The results showed that the plasmid size from pD353 was 34,063 bp, the content of G-C mol was 30.42%.Plasmid sequence analysis found that pD353 plasmid and Staphylococcus aureus plasmid sequence pIT1-S and pIT4-R had a high homologous, which was rich in heavy metal transport protein and insertion sequence translocation enzyme, its rep protein and pSK639 family plasmids had a high homologous. 【Conclusion】 Series repeat sequence analysis initially concluds that the plasmid is a θ replication and antibiotic resistance gene is unfound.