ANNOUNCEMENT {#s1}
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Agrobacterium tumefaciens C58 is a Gram-negative rod-shaped bacterium ubiquitously found in soil ([@B1]). As a plant pathogen, A. tumefaciens C58 contains plasmid TiC58 that transfers transfer DNA (T-DNA) to 90 families of dicotyledonous plants, inevitably resulting in crown gall tumors. Bacteriophages may be useful in manipulating this characteristic for engineering *Agrobacterium* strains. Here, we present the genome sequence of myophage Milano.

Milano was isolated from filtered rice stem extracts in Beaumont, TX, on A. tumefaciens C58 grown aerobically at 28°C in nutrient broth yeast (NBY) medium without glucose ([@B2]) by the soft agar overlay method ([@B3]). The high-titer lysate generated via the soft agar overlay method was used for extracting genomic DNA with the phenol-chloroform method, as in reference [@B4], and then the phage genomic DNA libraries were prepared using a NEBNext Ultra II DNA library prep kit and sequenced on an Illumina MiSeq instrument at the Genome Sequencing and Analysis Facility at the University of Texas at Austin ([@B5]). The 820,950 250-bp paired-end sequence reads were quality controlled with FastQC (<http://www.bioinformatics.babraham.ac.uk/projects/fastqc/>) and trimmed with the FASTX-Toolkit 0.0.14 ([hannonlab.cshl.edu/fastx_toolkit/](http://hannonlab.cshl.edu/fastx_toolkit/)). Complete assembly into a single contig using SPAdes v.3.5.0, with default parameters, was confirmed via PCR of the genome ends (forward primer, 5′-GTCCTGAATCCATTTCGTATGC-3′; reverse primer, 5′-CTCCGTTCTTCAGCTACTATG-3′), coupled with Sanger sequencing ([@B5], [@B6]). Genes were called and annotated using GLIMMER 3.0 and MetaGeneAnnotator 1.0 in the Web Apollo instance hosted by the Center for Phage Technology (<https://cpt.tamu.edu/galaxy-pub>), and all analyses were performed in their Galaxy instance ([@B7][@B8][@B10]). Potential tRNA genes were inspected using ARAGORN 2.36 ([@B11]). Gene functions were predicted using domains from InterProScan v.5.22, LipoP, and TMHMM, as well as BLASTp comparisons to the NCBI nonredundant (nr) and UniProtKB Swiss-Prot/TrEMBL databases ([@B12][@B13][@B16]). HHpred results were used as confirmatory evidence, in addition to the presence of domains or alignments ([@B17]). Rho-independent termination sites were detected using TransTerm (<http://transterm.cbcb.umd.edu/>). Milano's morphology was determined by negative-stain transmission electron microscopy at the Texas A&M Microscopy and Imaging Center with 2% (wt/vol) uranyl acetate ([@B18]).

Milano is a myophage with a 68,451-bp genome, 93.1% coding density, and a G+C content of 52.5%, which is lower than the G+C content of 58% of the host, A. tumefaciens C58 ([@B1]). Our analysis revealed 127 coding sequences, of which 45 have a function predicted by using BLASTp or InterProScan, but no tRNAs. Based on PhageTerm prediction, Milano uses headful DNA packaging ([@B19]). While Milano contains mostly hypothetical proteins, there is one closely related phage in the NCBI nr database, namely, *Agrobacterium* myophage 7-7-1 (GenBank accession number [JQ312117](https://www.ncbi.nlm.nih.gov/nuccore/JQ312117)) ([@B20]). By comparison with progressiveMauve, Milano has 61.68% nucleotide identity and shares 94 proteins with *Agrobacterium* phage 7-7-1, a flagellotropic phage ([@B21]).

The Milano genome is organized in a modular fashion, with predicted structural, replication, and lysis proteins grouped together. The predicted tape measure protein (GenBank accession number [QBQ72047](https://www.ncbi.nlm.nih.gov/protein/QBQ72047)) is preceded by the likely tail assembly chaperones ([QBQ72045](https://www.ncbi.nlm.nih.gov/protein/QBQ72045)) with a putative frameshifted protein product ([QBQ72046](https://www.ncbi.nlm.nih.gov/protein/QBQ72046)), analogous to the well-studied lambda G/GT chaperone system ([@B22]). The predicted endolysin ([QBQ72055](https://www.ncbi.nlm.nih.gov/protein/QBQ72055)), i-spanin ([QBQ72056](https://www.ncbi.nlm.nih.gov/protein/QBQ72056)), and embedded o-spanin ([QCQ78506](https://www.ncbi.nlm.nih.gov/protein/QCQ78506)) are encoded consecutively; however, the holin was not identified. Additionally, a putative nucleoid occlusion-like protein ([QBQ72073](https://www.ncbi.nlm.nih.gov/protein/QBQ72073)) with a ParB domain (InterProScan IPR003115) and many BLASTp hits to Noc proteins and two potential ribosome modulation factor domain superfamily proteins ([QBQ72082](https://www.ncbi.nlm.nih.gov/protein/QBQ72082) and [QBQ72098](https://www.ncbi.nlm.nih.gov/protein/QBQ72098)) were found.

Data availability. {#s1.1}
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The genome sequence and associated data for phage Milano were deposited under GenBank accession number [MK637516](https://www.ncbi.nlm.nih.gov/nuccore/MK637516), BioProject accession number [PRJNA222858](https://www.ncbi.nlm.nih.gov/bioproject/PRJNA222858), SRA accession number [SRR8869236](https://www.ncbi.nlm.nih.gov/sra/SRR8869236), and BioSample accession number [SAMN11360273](https://www.ncbi.nlm.nih.gov/biosample/SAMN11360273).

This work was supported primarily with funding from the National Science Foundation (awards EF-0949351 and DBI-1565146) and the Texas A&M AgriLife Research Zebra Chip Disease Program (grant number ZC 06-407697) to C.F.G. The Center for Phage Technology received an Initial University Multidisciplinary Research Initiative from Texas A&M University and Texas AgriLife.

We are grateful to the Texas A&M Department of Biochemistry and Biophysics and the CPT staff.

This announcement was prepared in partial fulfillment of the requirements for BICH464 Bacteriophage Genomics, an undergraduate course at Texas A&M University.

[^1]: **Citation** Nittolo T, Ravindran A, Gonzalez CF, Ramsey J. 2019. Complete genome sequence of *Agrobacterium tumefaciens* myophage Milano. Microbiol Resour Announc 8:e00587-19. <https://doi.org/10.1128/MRA.00587-19>.
