Pantoea Ananatis is a promising organism in biotechnology as well as for biological control because of its valuable enzymatic and antimicrobial bioactivities. In this study, we adopted a comparative approach to assess the genome sequence of Pantoea Ananatis 140- UN, which was derived from the digestive tract of Pintomyia evansi, a hematophagous insect. We further evaluated genomic and taxonomic variabilities and the relationship of gene regions with previously characterized in vitro bioactivities.The scaffold-level genome of P. ananatis 140-UN showed a total length of 4,929.135 bp, and 4,690 attributes were annotated. Furthermore, the genome was mainly composed of a chromosome and a plasmid (i.e., LPP-1); plasmid size was 308.225 bp, and G + C content was 53.2%. P. ananatis 140-UN was found to be closely related to SGAir0210, a P. ananatis strain isolated from outdoor air collected in Singapore. Pan-genomic analyses revealed regions that have not been identified in other previously described genomes. Besides, genomic analysis of antimicrobial resistance factors led to the identification of genes conferring resistance to aminoglycosides. Unique regions were also identified, corresponding to 888 genes (373 unique genes), which were mainly associated with genetic information processing and signaling and cellular processes.