GTF/GFF3 Sequence Extractor

A precise tool for extracting sequences from genome assemblies using GTF/GFF3 annotations, targeting exons, CDS, or entire gene regions.

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About This Tool

This tool retrieves nucleotide sequences from a genome assembly by applying a GTF/GFF3 annotation. It supports extracting exons, CDS, genes, or transcript regions, with optional strand-aware orientation and translation. Researchers supply a genome FASTA and an annotation file; the tool validates coordinates, resolves feature boundaries, and outputs a FASTA file with one sequence per requested feature. It is designed for reproducible workflows and can integrate into pipelines that require feature-specific sequences for primer design, alignment, or comparative analyses.
Conceptually, the tool parses the annotation, maps features to genomic coordinates, orders components (e.g., exons within a transcript), and concatenates sequences according to the feature type. On the negative strand, sequences are reverse-complemented when requested. Optional extensions allow upstream/downstream sequence context and translation of CDS to protein using the standard genetic code. Outputs can include metadata reports in CSV or BED formats to support downstream processing.
The primary beneficiaries are genomics researchers, bioinformaticians, and core facilities who need reliable, repeatable sequence retrieval for discovery, validation, or pipeline automation. By separating feature resolution, sequence extraction, and optional translation, the tool provides a clear, auditable trace of how each sequence was derived and how downstream analyses were generated.

How to Use

  1. Provide inputs: upload genome_fasta and gtf_gff3 files.
  2. Choose feature_type (exon, CDS, gene) and, if desired, feature_ids or region_filter.
  3. Configure optional parameters: strand, upstream/downstream extension, translation, output_format.
  4. Run extraction.
  5. Download and review extracted_sequences FASTA and any reports.
How to use gtf/gff3 sequence extractor

Frequently Asked Questions

Find Quick Answers

What formats are supported for inputs and outputs?
Inputs require FASTA for the genome and GTF or GFF3 for annotations. Outputs are provided as FASTA sequences; optional CSV or BED metadata may be generated for integration with other tools.
Can CDS be translated to protein sequences?
Yes. If translation is enabled, CDS-derived sequences are translated using the standard genetic code to produce protein sequences in a separate FASTA output.
How are exons concatenated or CDS assembled?
Exons are ordered by transcript or gene structure from 5' to 3' on the strand; CDS takes all CDS features in transcription order. Regions on the negative strand are reverse-complemented when extracted.
What if annotation coordinates do not match the genome FASTA?
The tool detects mismatches and reports warnings. Extraction proceeds only for features with valid, in-range coordinates; invalid features are skipped with a summary.

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