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Reference open reading frames and experimental translation evidence on PyEnsembl transcripts

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orfdata

orfdata builds the protein reference for a species: the proteins annotated in a PyEnsembl genome, plus open reading frames (ORFs) that ribosome-profiling studies found translated outside those annotations. These include upstream and downstream ORFs, ORFs in other reading frames of a coding sequence, and ORFs on RNAs annotated as noncoding. Each added ORF stays bound to its PyEnsembl transcript, with its source, category and evidence.

Installation

Use Python 3.9 or later. Install orfdata and one PyEnsembl annotation:

python -m pip install 'orfdata[pyensembl]'
pyensembl install --release 115 --species human

Example

from pyensembl import EnsemblRelease
from orfdata import Proteome

genome = EnsemblRelease(115, species="human")
proteome = Proteome.from_genome(genome).with_extra_orfs(evidence="ms")
proteome.write_fasta("human-e115.fasta")
print(proteome.summary()["kinds"], len(proteome.excluded))
{'annotated': 245535, 'extra_orf': 2181} 5

This takes every protein in the Ensembl 115 annotation and adds the 2,181 bundled human ORFs that have supporting mass-spectrometry peptides and are encoded by an Ensembl 115 transcript. Each entry lists every transcript encoding it; the other 5 selected ORFs are listed in proteome.excluded with the reason. Use evidence="ribosome" for all 7,239 encoded ribosome-profiling ORFs, or a rule such as RiboSeq(min_studies=2) & MS() to set Ribo-seq and MS stringency separately (1,293 encoded ORFs). proteome.save(path) writes a checksummed snapshot that Proteome.load reads offline. From the command line:

orfdata proteome --species human --release 115 --output human-e115.fasta

A translated ORF or a detected peptide does not by itself establish a stable, functional protein. Most MS support in the bundled human catalogue comes from immunopeptidomics: peptides eluted from HLA molecules.

Species

Species Noncanonical ORFs Sources
Human 7,264 bundled; 57,607 with catalogs="all" GENCODE; optionally Chang et al. 2025 and Yang et al. 2024 Ribo-seq
Mouse 48,805 Chang et al. and Yang et al. Ribo-seq, OpenProt 2.2 MS
Zebrafish 9,759 Yang et al. Ribo-seq
C. elegans 2,849 Yang et al. Ribo-seq
Yeast 1,128 Yang et al. Ribo-seq
Dog none: no public dog Ribo-seq or ncORF MS exists

Human's bundled catalogue needs no setup. For the others, pip install "orfdata[data,build-data]", then orfdata setup --species mouse (or zebrafish, worm, yeast, or human --catalogs all) builds the catalogues from their sources' own files (details). After orfdata chains download (add --accept-terms for UCSC's non-commercial mouse chains), older assemblies (GRCh37, NCBI36, GRCm38, NCBIM37) and T2T-CHM13 use these catalogues lifted to them; orfdata genomes install t2t-chm13 installs a CHM13 annotation. Dog proteomes contain their PyEnsembl-annotated proteins. load_catalog(species=...) and with_extra_orfs() pick the catalogue for a species and raise when none is available or built. Human evidence is never transferred to another species. Viral proteomes can be added with with_viral(...), but only to a proteome for one of the virus's recorded hosts. orfdata also provides RNA, ribosome and protein measurements for human, mouse, dog and yeast.

viral_references(host="human", group="HPV", virus_type="HPV16") finds source-qualified ViralReference objects. Choose a stable ID with viral_reference("NC_001526.4"), then use .dna and .orfs to load its packaged bases and coding paths. The functional loaders remain available. When annotating an existing alignment, viral_orfs(reference_dna, contig="HPV16") checks its actual reference sequence before applying the same models. The first coding reference contains the nine RefSeq NC_001526.4 HPV16 CDSs, with native and GDC placements; the host association is distinct from the virus's own taxon. Broader imports remain in the viral roadmap.

Measurement datasets

RNA, ribosome and protein measurements are separate, downloadable datasets with the same species-scoped API. Install the data extra, then download and query one:

pip install 'orfdata[data]'
orfdata datasets list
orfdata datasets download ingolia-yeast-rna-ribo
Guide Species Measurements
Protein abundance Human, mouse, dog TMT protein groups (Oziolor et al.)
RNA abundance Dog, mouse Native gene counts (Oziolor et al.)
Matched human tissues Human RNA and protein from adjacent cryosections (Wang et al.)
Yeast protein and RNA Budding yeast Calibrated protein and RNA abundance (CoPY)
Yeast RNA and ribosome occupancy Budding yeast Source regions, including UTR and ncRNA labels (Ingolia et al.)
Dog peptides Dog DLA class I ligands from tumours (Kaabinejadian et al.); Canine PeptideAtlas peptides and protein calls, built locally

Gene RNA and canonical protein measurements do not establish noncanonical ORF abundance. See released data and gaps.

Documentation

The documentation covers:

Task Guide
Build a proteome: evidence levels, exclusions, contigs, viral proteins Build a reference proteome
ORF classes, evidence definitions, Varcode and Isovar integration ORFs and evidence
Species and reference identity, mitochondrial genetic codes Species, references and genetic codes
Versioned ORF catalogues managed through DataCache Optional ORF catalogues
Quantitative datasets and what has been released Measurements, released data and gaps

The data roadmap tracks further species and sources. Every PR includes a version bump, lint and tests; maintainers publish with ./deploy.sh from clean main.

License

Apache 2.0. Downloadable datasets keep their sources' own terms and attribution, recorded in each release manifest.

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Reference open reading frames and experimental translation evidence on PyEnsembl transcripts

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