The authors declare that there are no conflicts of interest present.
Microbacteriophage Babydotz was isolated in Moorhead, Minnesota in 2020 by infecting
TEM of uranyl acetate-stained microbacteriophage Babydotz shows a siphovirus morphology.
Research on bacteriophages has contributed to our understanding of viruses more broadly and in their development as therapeutics for multi-drug resistant bacterial infections (Kim et al, 2025). Here, we report the genome sequence of Babydotz, a microbacteriophage that infects
Babydotz was isolated from a soil sample that was collected between two pine trees at the Minnesota State University Moorhead Campus Mall (global positioning system [GPS] 46.8673 N, 96.762 W) when the ambient temperature was -2°C. Briefly, Babydotz was extracted by washing the soil with PYCa liquid medium, filtering the wash, and plating the filtrate in top agar with
DNA was extracted from a Babydotz lysate using a Promega Wizard DNA extraction kit prior to sequencing. The Pittsburgh Bacteriophage Institute generated sequencing libraries using the NEB
Auto-annotation was performed using DNA Master v5.23.6 (Pope et al., 2018) which employs GeneMark v.3.02 (Delcher et al., 1990) and Glimmer v.2.5p (Borodovsky et al., 2003; Besemer et al., 2005). DNA Master, Phamerator, using Actino_draft database v578 (Cresawn et al., 2011) and PECAAN (Rinehart et al., 2016) were used to manually refine the annotations. Starterator (Pacey, 2014) was used to further evaluate start sites conservation with Glimmer and Genemark. When there was disagreement among these software programs, BLAST results, coding potential and gene length were also used to determine the most likely start for the gene. No tRNA coding genes were identified using tRNAscan-SE v2.0 (Lowe, 2016) or ARAGORN v1.2.41 (Laslett, 2004). Putative functions for 23 of 104 predicted coding ORFs were assigned using BLASTp with an e-value cutoff of 10
-4
, using the Actinobacteriophage and NCBI non-redundant database (Altschul et al., 1990) and HHPRED, using the PDB_mmCIF70, Pfam- v.36, NCBI Conserved Domains databases with a probability of 90% or greater (Zimmermann et al., 2018; Söding et al., 2005). Nine protein-coding ORFs with membrane-associated domains were determined using SOSUI (Hirokawa et al., 1998) and TMHMM v.1.0.24 (Hallgren et al., 2022). All tools were run with default parameters. Following student genome annotation, the annotation underwent peer review by an experienced SEAPhage-associated faculty and passed quality control inspection. AUG start codon usage was 100%. BabyDotz was assigned to cluster EG based on gene content similarity of at least 35% to phages in the Actinobacteriophage database, phagesdb (
Other predicted genes include those encoding a putative DprA-like DNA processing chain A, three helix-turn-helix DNA binding domains, a terminase, a portal protein, a major capsid protein, a major tail protein, a phosphoesterase, a head-to-tail adaptor, a tape measure protein, three minor tail proteins, an endolysin, an exonuclease, an HNH endonuclease, a RuvC-like resolvase, a MazG-like nucleotide pyrophosphohydrolase, an SSB protein, a DNA primase/helicase and two ribbon-helix-helix DNA binding proteins. No genes encoding immunity repressor or integrase functions could be identified, suggesting that Babydotz is unlikely to establish lysogeny.
Babydotz is available at GenBank with Accession No. 0R613478 and Sequence Read Archive (SRA) No. SRR18306109.
We thank The Howard Hughes Medical Institute and the Hatfull Lab at the University of Pittsburgh for supporting this research. We would also like to thank Ethan Dotzler and Jonathan Nyandu Kanyinda and their supporting faculty from Minnesota State University Moorhead for the discovery, purification, isolation of DNA from and transmission electron microscopy of phage Babydotz.