Supporting dataset: In silico primer design and validation for five biotechnological marker genes — primer sequences, thermodynamic parameters, design configurations, and Primer-BLAST specificity results

This dataset contains all primary data supporting the manuscript "A reproducible pipeline for rational primer design and in silico validation of biotechnological marker genes" (submitted to Acta Scientiarum. Biological Sciences). Contents:— primer_dataset_zenodo.xlsx: Six-sheet Excel workbook with primer sequences, thermodynamic parameters (Tm, %GC, self-complementarity ΔG, 3′-end ΔG, heterodimer ΔG), design configurations per gene, reference sequence metadata, Primer-BLAST specificity results, and confirmed amplicon sizes.— primers_all_genes.fasta: All 10 primer sequences in annotated FASTA format, independently verifiable with any oligonucleotide analysis tool.— screenshots_oligo_explorer/: Configuration and results screenshots for luxA, lacZ, spo0A, and ACT1 in Oligo Explorer v1.2.— screenshots_primer3plus/: Configuration and results screenshots for lasR in Primer3Plus v3.2.4.— snapgene_files/: Primer binding position screenshots (SnapGene v6.2.2) for all five genes and virtual gel electrophoresis output (2.0% agarose). Target genes: lacZ (E. coli K-12), spo0A (B. subtilis str. 168), ACT1 (S. cerevisiae S288C), lasR (P. aeruginosa PAO1), luxA (A. fischeri ES114).Tools: Oligo Explorer v1.2, Primer3Plus v3.2.4, OligoAnalyzer, NCBI Primer-BLAST (BLAST+ v2.15.0), SnapGene v6.2.2.Primer-BLAST searches conducted July 2025 against RefSeq Representative Genomes and NCBI nt collection.