Whole-Genome Sequencing and Annotation of KFS-EC3 Phage

The genomic DNA of KFS-EC3 was extracted and purified using a Phage DNA Isolation Kit (Norgen Biotek Corp. Thorold, ON, Canada). Whole-genome sequencing of the purified DNA was performed (Macrogen Inc., Seoul, Korea) using the paired-end Miseq sequencing platform (Illumina Inc., San Diego, CA, USA). The raw reads were trimmed using Trimmomatic [28 (link)] to eliminate the low-quality reads and adapter sequences. The de novo assembly of the qualified sequences was performed by various k-mer using the SPAdes genome assembler (Illumina). The open reading frames (ORFs) of the assembled sequence were predicted and annotated using the Rapid Annotations using Subsystems Technology (RAST) server [29 (link)] and BLASTP. The potential tRNA genes in the genome sequence were predicted using ARAGORN [30 (link)] and RNAmmer (version 1.2) [31 (link)] in the rapid prokaryotic genome annotation pipeline (Prokka) [32 (link)]. The complete genome sequence of KFS-EC3 was deposited in the GenBank database, under the nucleotide sequence accession number MZ065353. The novelty of phage was assessed based on its similarity to other most closely related phages from the NCBI BLASTN database (https://blast.ncbi.nlm.nih.gov/Blast.cgi?PAGE_TYPE=BlastSearch, 5 May 2021).