Purpose The purpose of this study was to assess the prevalence and associated factors of myopia among medical students at the University of Gondar College of Medicine and Health Sciences, Northwest Ethiopia. Patients and Methods Institution-based cross-sectional study was conducted at the University of Gondar College of Medicine and Health Sciences from June 20 to August 15, 2021. A simple random sampling technique with proportional allocation was used to select 492 students. Interviewer-administered questionnaire, Snellen acuity chart, pinhole, retinoscope and direct ophthalmoscope were used to collect the data. The data were entered to EpiData version 4.6 and exported to SPSS version 25 for analysis. Descriptive statistics were summarized by measures of central tendency. Variables with p-value ≤0.2 in bivariable regression were entered into a multivariable logistic regression model. Adjusted odds ratio with 95% confidence interval was used and variables with p-values <0.05 were considered as statistically significant. Results A total of 492 respondents participated giving a response rate of 98.3%. The prevalence of myopia was found to be 16.7% (95% CI = 12.8–19.4%). Urban residents (AOR = 1.56; 95% CI: 1.28–6.21), family history of myopia (AOR = 2.31; 95% CI: 1.33–4.54), near-work activity of 5–7 hours (AOR = 2.41; 95% CI: 1.31–5.76) and ≥8 hours (AOR = 4.35; 95% CI: 1.96–9.66), outdoor activity for <3 hours (AOR = 1.65 95% CI: (1.14–4.53), were significantly associated with myopia. Conclusion The prevalence of myopia among medical students at the University of Gondar College of Medicine and Health Sciences was high. Urban residency, positive family history, longer time spent on near-work activities and less outdoor activity were positively associated with myopia.