【Abstract】 Objective To explore the genetic information of phage 33D and its antituberculosis potentials. Methods The genome of phage 33D were sequenced. The gene annotations of those genomes were completed by DNAStar, TRF, Promoter predictions, tRNAscanSE, Glimmer and BLAST. Results The genome of phage 33D was a dsDNA sequence of 50036 bp. The genome of phage 33D encodes 84 gene, 17 gene functions had been identified by BLAST comparison. We propose that gene47 and gene 48 could encode lyases. We proposed that gene6, 7, 8, 9 and 14 were related to DNA Replication . Conclusion 33D is lytic phage which have potentials in control of tuberculosis and have not virulence gene. Phage 33D could be used in phage cocktail therapy.