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Objectives: To estimate the costs to implement public health department (PHD)-run COVID-19 vaccination clinics., Design: Retrospectively reported data on COVID-19 vaccination clinic characteristics and resources used during a high-demand day in March 2021. These resources were combined with national average wages, supply costs, and facility costs to estimate the operational cost and start-up cost of clinics., Setting: Thirty-four PHD-run COVID-19 vaccination clinics across 8 states and 1 metropolitan statistical area., Participants: Clinic managers at 34 PHD-run COVID-19 vaccination clinics., Intervention: Large-scale COVID-19 vaccination clinics were implemented by public health agencies as part of the pandemic response., Main Outcomes Measured: Operational cost per day, operational cost per vaccination, start-up cost per clinic., Results: Median operational cost per day for a clinic was $10 314 (range, $637-$95 163) and median cost per vaccination was $38 (range, $9-$206). There was a large range of operational costs across clinics. Clinics used an average of 99 total staff hours per 100 patients vaccinated. Median start-up cost per clinic was $15 348 (range, $1 409-$165 190)., Conclusions: Results show that clinics require a large range of resources to meet the high throughput needs of the COVID-19 pandemic response. Estimating the costs of PHD-run vaccination clinics for the pandemic response is essential for ensuring that resources are available for clinic success. If clinics are not adequately supported, they may stop functioning, which would slow the pandemic response if no other setting or approach is possible., Competing Interests: Conflict of Interest: None., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)