Francisella tularensis, the causative agent for tularaemia, is a Tier 1 select agent, and a pan-species pathogen of global significance due to its zoonotic potential. Consistent genome characterization of the pathogen is essential to identify novel genes, virulence factors, antimicrobial resistance genes, for studying phylogenetics and other features of interest. This study was conducted to understand the genetic variations among genomes ofF. tularensisisolated from two felines and one human source. Pan-genome analysis revealed that 97.7 % of genes were part of the core genome. All threeF. tularensisisolates were assigned to sequence type A based on single nucleotide polymorphisms (SNPs) insdhA. Most of the virulence genes were part of the core genome. An antibiotic resistance gene coding for class A beta-lactamase was detected in all three isolates. Phylogenetic analysis showed that these isolates clustered with other isolates reported from Central and South-Central USA. Assessment of large sets of theF. tularensisgenome sequences is essential in understanding pathogen dynamics, geographical distribution and potential zoonotic implications.