The beloved country music star, 75, broke into song while getting the jab and adapted one of her best-known ballads.
To the tune of Jolene, Parton sang: “Vaccine, vaccine, vaccine, vaccine, I’m begging of you, please don’t hesitate.
“Vaccine, vaccine, vaccine, vaccine, because once you’re dead, then that’s a bit too late.”
Parton was credited with helping fund the Moderna vaccine after donating one million dollars (about £716,000) to Vanderbilt University Medical Centre in Nashville, Tennessee.
“I’m so excited. I’ve been waiting a while,” the singer told fans in a video posted from the university. “I’m old enough to get it and I’m smart enough to get it.
“I’m trying to be funny now, but I’m dead serious about the vaccine. I think we all want to get back to normal, whatever that is. And that would be a great shot in the arm, wouldn’t it, if we could get back to that?
“But anyhow, I just wanted to encourage everybody because the sooner we get to feeling better, the sooner we are going to get back to being normal.
Parton added: “I just want to say to all of you cowards out there - don’t be such a chicken squat. Get out there and get your shot.”
Parton masked up before inviting a doctor into the room. The medic, an old friend of Parton, praised her for urging others to get the vaccine.
After he knocked an item off the table while preparing the jab, a giggling Parton quipped: “I didn’t know you was going to be so clumsy, I hope you’re going to do better with my shot!”
After getting her first jab, Parton said: “That didn’t hurt.”
Parton is the latest celebrity to publicly confirm they have had the vaccine.
Sir Elton John, Sir Michael Caine, Sir David Attenborough, Sir Tony Robinson and Sir Ian McKellen have all had the jab.