gORF_25784 microprotein

Same-strand overlap (alternative frame) existence proven (MS) function unknown 58 aa

A non-canonical small protein (smORF) excluded from the H37Rv annotation, detected by proteogenomics with a high-confidence peptide-spectrum match. It is served here as part of a separate microproteome track: it is not one of the 3906 canonical genes and does not count in the dark stock. Its existence is proven; its molecular function is unknown. It lies on the same strand within pflA, i.e. an alternative reading frame of a known gene (dual coding) rather than a new locus.

Evidence & coordinates

Detection method	MS peptide-spectrum (compendium)
Existence evidence	high-confidence peptide-spectrum match (Spectrum Forest hc) (tier T1)
Cross-validation	no Ribo-RET footprint at this start (Smith et al. 2022). Not informative on its own: measured genome-wide, this Ribo-RET dataset recovers only 29.1 % of annotated H37Rv starts (1136/3906), so its absence here does not argue against this microprotein — existence is already established by the MS evidence above. (P20.5)
H37Rv coordinates	3504165–3504227 (+ strand)
Length	58 amino acids
Shine-Dalgarno	Present. Low to moderate binding.
Folding free energy	-3.49838
Predicted localisation	Membrane (2.191)
Conservation	homologues detected in Actinobacteria, Mycobacteria
Overlaps locus	pflA (Rv3138) — by genomic geometry

Sequence

MIVKSAADETQATLSLVKFACAWTYIALDPAVVQSVPVFGCDGERVAHGALERQVTRR

Source: de Souza EV et al., Sci Rep 2024;14:31186 (doi:10.1038/s41598-024-82465-w); proteogenomics microprotein compendium. Overlap class is assigned by geometry against the H37Rv CDS set, not by the source's own label. ← all microproteins