Genomic Coordinates

When to use

Any time a coordinate crosses a boundary: between two file formats, between two tools, between two assemblies, or between the genome and a transcript.

The rule

A coordinate is three facts, not one: the number, the convention it is written in, and the assembly it was measured against. Carry all three or the number is not interpretable.

Coordinate errors are the quietest class of bug in genomics. An off-by-one BED file parses, sorts, and intersects without complaint. A GRCh37 VCF joined against a GRCh38 annotation returns rows. A right-shifted indel simply fails to match its entry in ClinVar, and the result is a variant reported as novel. Nothing raises an error; the answer is just wrong, and it is wrong in a direction that looks plausible.

So: convert with the table, not from memory, and verify against the reference whenever a reference is available.

The two conversions

1-based inclusive  ->  0-based half-open :  start - 1,  end
0-based half-open  ->  1-based inclusive :  start + 1,  end

These formulas apply to nonempty spans on the same reference and strand. They do not encode insertions, circular wraparound, liftover, or transcript mapping.

Which format is which

Both "MAF" formats exist, they mean different things, and they disagree. UCSC serves 0-based files through a 1-based browser box. references/format-conventions.md has the full table with per-format detail.

cd skills/genomic-coordinates/scripts

python3 convert_coords.py --list                          # the table
python3 convert_coords.py --from bed --to gff chr1 999 1000
python3 convert_coords.py --from ucsc --to bed "chr7:5,530,601-5,530,625"
python3 convert_coords.py --from granges --to pyranges --input regions.tsv

contig  input                 output           length  status  detail
chr7    chr7:5530601-5530625  5530600-5530625  25      ok

Zero-length BED features (chromStart == chromEnd, a legal insertion point) are reported as unrepresentable for an inclusive target because the converter lacks feature semantics. GFF3 can encode insertion sites with equal endpoints and a feature type; that is different from an ordinary single-base interval. Exit code is 1 for invalid or unrepresentable output; valid zero-length half-open output exits 0. Output is a diagnostic TSV/JSON table, not a rewritten GFF/VCF.

--input parses BED/bedGraph, GFF/GTF, literal-allele VCF REF spans, explicit region strings, and three-column GRanges/PyRanges/Python TSVs. Other table rows are conventions only: extract an interval with a native parser and pass a triple. All bundled text readers expect uncompressed files.

Variants are not intervals

For a simple VCF indel, POS normally identifies the unchanged padding base before the event. At contig position 1 the padding can follow the event. Complex substitutions need not have an unchanged anchor. And the same change can be written many ways: chr1:7:CAC:C, chr1:3:CAC:C and chr1:2:GCA:G are one deletion. Joining, deduplicating, or looking up variants before normalising loses real matches silently, and it loses them preferentially in repeats, where indels concentrate.

Normalise — trim to parsimony, then left-align against the reference — before any comparison:

python3 normalize_variant.py --fasta ref.fa chr1 7 CAC C
python3 normalize_variant.py --fasta ref.fa --split --input cohort.vcf
python3 normalize_variant.py --fasta ref.fa --compare chr1:7:CAC:C chr1:2:GCA:G

input         normalized    type      pos_shift  ref_check  changed
chr1:7:CAC:C  chr1:2:GCA:G  deletion  5          ok         yes

Literal alleles are checked against the FASTA using exact contig names. A MISMATCH can indicate an assembly, sequence, strand, or coordinate error; stop and investigate with check_contigs.py and sequence provenance. The helper rejects unsplit ALT lists: use --split for independently normalized allele keys. Its TSV discards genotypes and annotations; use bcftools norm for production VCF rewriting. Symbolic/breakend/missing/spanning-deletion alleles are passed through as skipped, without REF or structural validation.

The default left-shift window is 1,000 bp. If it prevents completion, the helper returns incomplete, exits 1, and refuses an equivalence verdict. Increase --window and rerun. Matching normalized keys tests individual literal alleles, not haplotype equivalence across multiple records.

HGVS applies the 3'-most rule to the reference sequence being described. For transcript c./n. notation, this means increasing genomic coordinates on a plus-strand gene and decreasing coordinates on a minus-strand gene. The minus-strand direction can therefore agree with VCF left-alignment; genomic g. notation shifts toward the contig end. Details and exceptions: references/variant-representation.md.

Check the assembly before trusting a join

python3 check_contigs.py --identify unknown.fa.fai
python3 check_contigs.py variants.vcf annotation.gtf --genome GRCh38.fa.fai

file          kind    contigs  naming        assembly  detail
ref.fa.fai    sizes   25       plain         GRCh37    24/24 primary chromosome lengths match;
                                                       chrM is 16569 bp, i.e. GRCh37/38 (rCRS MT)

The script reads .fai, .chrom.sizes, VCF headers, SAM headers, FASTA, BED, and GTF/GFF, identifies the assembly from primary-chromosome lengths, and reports detectable conflicts: naming mismatch, length conflict, coordinates past a contig end, contigs present in one file only. Exit code 1 on a detected conflict. unknown/ambiguous with exit 0 is not proof of compatibility; lengths cannot detect same-length sequence changes or masking. Reference contig supersets are expected. VCF header and record extents are both checked when comparing; SV/gVCF spans require a native validator.

GRCh37 and hg19 share primary nuclear coordinates, but differ in mitochondrial reference — 16,569 bp (rCRS) versus 16,571 bp. Nuclear coordinates are identical, so a mixed pipeline runs fine and only the mtDNA results are wrong. check_contigs.py reports which one it found. Builds, naming schemes, ALT contigs, and liftover pitfalls: references/reference-builds.md.

Audit a file against its own format

python3 audit_intervals.py peaks.bed
python3 audit_intervals.py gencode.gtf --genome hg38.chrom.sizes
python3 audit_intervals.py cohort.vcf --genome GRCh38.fa.fai

Looks for the evidence that a coordinate mistake leaves behind:

Exit code 1 on any fatal finding. This is a targeted coordinate audit, not a full format validator. Special VCF alleles produce structural_extent_unchecked; circular GFF3 spans need feature-aware validation. The narrowPeak/broadPeak readers do not interpret signal columns as BED thickStart/thickEnd.

Transcript, CDS, and protein positions

c.742 and chr17:7,674,220 are both "position", and neither converts to the other by arithmetic. Transcript coordinates count spliced bases in transcription order — decreasing genomic coordinate on the minus strand — and c.1 is the A of the initiator ATG, not the start of the transcript.

The rules that get mis-remembered: there is no c.0; 5' UTR positions are negative and 3' UTR positions take a *; GFF phase counts bases to skip when locating the next complete codon within a CDS segment (retain them when joining coding exons), not start % 3; and a c. description is meaningless without a versioned transcript accession, because the same variant numbers differently in each transcript. references/transcript-coordinates.md has the conversion procedure and the boundary cases.

Use VEP, Mutalyzer, or the hgvs package with the matching transcript model for HGVS conversion. bcftools csq annotates haplotype-aware coding effects; it is not a general genomic-to-HGVS converter.

Reporting results

State the assembly next to the coordinates, every time. chr7:5,530,601-5,530,625 is not a location; chr7:5,530,601-5,530,625 (GRCh38) is. Say which convention a coordinate column is in, in the column header or the file's documentation. When a conversion produced a result, say which direction it went.

Verified scope

Reviewed the current VCF 4.5, SAM/BAM, CRAM 3, GFF3, UCSC, HGVS, Ensembl REST, and bcftools manuals on 2026-10-01. Bundled standard-library helpers are tested on synthetic fixtures; normalization is cross-checked against bcftools 1.24. Transcript annotation and liftover tools are documented alternatives, not executed whole-genome workflows. Source links are in the references below.

References

• references/format-conventions.md — every format's convention, with per-format detail, BED12 block rules, region-string syntax, and tool behaviour.
• references/variant-representation.md — VCF allele conventions, the normalisation algorithm, equivalence checking, multi-allelic splitting, and how HGVS disagrees with VCF.
• references/reference-builds.md — build signatures, GRCh37 vs hg19, ALT contigs, naming schemes, and liftover failure modes.
• references/transcript-coordinates.md — genomic ↔ transcript ↔ CDS ↔ protein, HGVS numbering, phase, and transcript choice.