EZsplit Instructions


EZsplit extracts mitochondrial protein-coding genes from annotated genome files and generates separate FASTA files for each gene.

  • Reads genomic sequences from a FASTA file
  • Parses gene coordinates from a GFF3 annotation file
  • Selects features annotated as protein_coding
  • Matches gene names to standard mitochondrial gene synonyms
  • Extracts gene sequences using annotation coordinates
  • Reverse-complements genes on the negative strand
  • Creates one FASTA file per gene
  • Generates a missing_genes.txt report
  1. Provide a genomic FASTA file (.fasta, .fa, .fna, .fas)
  2. Provide the corresponding GFF3 annotation file (.gff, .gff3)
  3. Ensure record IDs match between FASTA and GFF
  4. Only gene_biotype=protein_coding features are extracted
  5. Gene names must match mitochondrial gene standards

Upload Your Files


Drag & drop FASTA file
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Drag & drop GFF file
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