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FASTA format guide

Everything you need to create readable, valid FASTA files for DNA, RNA, and protein sequences.

Updated July 2026 · 4 min read

What is FASTA?

FASTA is a simple text format used to store biological sequences. Each record contains a description line, called a header, followed by one or more lines of sequence data. FASTA files commonly use the extensions .fasta, .fa, or .faa.

Quick rule: Every record must begin with a greater-than symbol (>) and a non-empty identifier. Keep sequence lines free of spaces and numbers.

Basic file structure

A minimal FASTA record has two parts. The first line starts with >; everything after it is the sequence label and optional description. The sequence begins on the next line.

>human_insulin_chain_b
FVNQHLCGSHLVEALYLVCGERGFFYTPKTRREAEDLQVGQVELGGGPGAGSLQPLALEGSLQ

A file can contain multiple records. Start each new record with its own header line.

>sample_alpha
ACGTACGTACGTACGT
>sample_beta
GGCCTTAAGGCC

Sequence alphabets

Use the alphabet that matches your sequence type. The generator uses these standard symbols when creating a random sequence.

TypeCommon symbolsTypical extension
DNAA, C, G, T.fasta or .fa
RNAA, C, G, U.fasta or .fna
Protein20 standard amino-acid letters.faa

Some tools also accept ambiguity codes such as N for an unknown nucleotide or X for an unknown amino acid. Check the requirements of the downstream tool before including them.

Best practices

  • Use short, unique labels without spaces. Underscores and hyphens are broadly compatible.
  • Wrap sequence lines at 60 characters for readability and compatibility with many command-line tools.
  • Keep one sequence record per logical sample, and avoid adding comments inside sequence data.
  • Save the file as plain text using UTF-8 encoding, not as a rich text document.
  • Verify that DNA uses T and RNA uses U before analysis.