The primary objective of this project was to generate the genomic sequence of the Naegleria fowleri strain Nf69, originally isolated from a human-infected brain in 1969 in Adelaide, Australia. This specific strain is deposited in the culture collection ATCC 30215 and is classified as belonging to the IV/5 genotype. This project involved sequencing and analyzing the genomic content of the Naegleria fowleri strain Nf69, leading to comparative genomic insights under different drug conditions and a detailed exploration of its non-coding repertoire. These findings contribute to a deeper comprehension of the strain's biology and pathogenicity, potentially offering new avenues for drug development and treatment strategies.