Background: Shoulder pain, a common reason for seeking physical therapy (PT), may arise from the cervical spine or shoulder. Symptom localization (SL), is a systematic process to determine the source of symptoms through provocation/ alleviation testing. Objectives: To illustrate the clinical application of the SL sequence in a patient with shoulder pain. Design: A retrospective case report. Methods: A 29-year-old, female presented to PT with a 6-month history of right anterolateral shoulder pain that began after leaning on her arm while driving. Symptoms were aggravated with overhead movement and alleviated by external support. A thorough history/medical screening was performed. The SL examination sequence included: regional localization, structural localization, and confirmation testing. Results: The shoulder was identified as the source of symptoms (supraspinatus muscle-tendon unit). Prognosis was good secondary to the absence of underlying comorbidities and the ability to manually and positionally alleviate symptoms. Intervention included a combination of manual therapy (functional massage and joint mobilization) and neuromuscular re-education (gravity reduced training progressed to resisted exercises). Conclusion: This case provides an in-context example of SL to determine the source of symptoms, allowing for the implementation of a phased treatment approach, and return to prior level of function.