Plants have long been at the main focus of the medical industry's attention due to their extensive list of biological and therapeutic properties and ethnobotanical applications. Catharanthus roseus, sometimes referred to as Nithyakalyani in Tamil, is an Apocynaceae family member used in traditional Indian medicine. It also examines the plant's potential antimicrobial and antioxidant activities as well as its preliminary phytochemical makeup. Leaf material from C. roseus was analyzed and found to include a variety of phytochemicals including alkaloids, terpenoids, flavonoids, tannins, phenols, saponins, glycosides, quinones, and steroids. Four of the seven secondary metabolic products discovered in C. roseus leaves showed bioactive principles: 3-methylmannoside, squalene, pentatriacontane, and 2,4,4-trimethyl-3-hydroxymethyl-5a-(3-methyl-but-2-enyl)-cyclohexene. Catharanthus roseus is rich in the anticancer compounds vinblastine and vincristine. Whole DNA was isolated from fresh leaves, then amplified, sequenced, and aligned to find prospective DNA barcode candidates. One DNA marker revealed the restricted genetic relationship among C. roseus based on genetic distance and phylogenetic analysis. The antioxidant activity of the plant extract was evaluated using the DPPH, ABTS, phosphomolybdenum, FRAP, and superoxide radical scavenging activity assays, while the antibacterial potential was evaluated using the agar well diffusion assay. The ethanol extract of C. roseus was found to have the highest reducing power. In addition, a 4- to 21-mm-wide zone of inhibition was seen when the C. roseus extract was tested against bacterial and fungal stains. In conclusion, C. roseus has the most promise as an antibacterial and antioxidant agent.