[22] In 1774, Paul Revere added Franklin's iconic cartoon to the nameplate of Isaiah Thomas's paper, the Massachusetts Spy, depicted there as fighting a British griffin.

She never begins an attack, nor, when once engaged, ever surrenders [...] The Rattle-Snake is solitary, and associates with her kind only when it is necessary for their preservation [...] 'Tis curious and amazing to observe how distinct and independent of each other the rattles of this animal are, and yet how firmly they are united together, so as never to be separated but by breaking them to pieces.

[...] The power of fascination attributed to her, by a generous construction, may be understood to mean, that those who consider the liberty and blessings which America affords, and once come over to her, never afterwards leave her, but spend their lives with her.The rattlesnake symbol was first officially adopted by the Continental Congress in 1778 when it approved the design for the seal of the War Office.

Continental Colonel Christopher Gadsden represented South Carolina in the Congress, and he was one of seven members of the Marine Committee outfitting the first naval mission.

"[25] The first Marines carried drums painted yellow and depicting a coiled rattlesnake with thirteen rattles along with the motto "Don't Tread on Me."

[26][6] He gave Commodore Esek Hopkins a yellow rattlesnake flag to serve as his personal standard on USS Alfred, the flagship of America's first navy squadron.

[28][29] He unfurled the Gadsden flag on the main mast of USS Alfred on December 20, 1775, while the ship was at anchor in Chesapeake Bay.

[5] Gadsden's presentation of the rattlesnake flag was recorded in the South Carolina congressional journals on February 9, 1776: Col. Gadsden presented to the Congress an elegant standard, such as is to be used by the commander in chief of the American Navy; being a yellow field, with a lively representation of a rattlesnake in the middle in the attitude of going to strike and these words underneath, "Don't tread on me.

"[31] In 1861, a ship from Georgia entered Boston Harbor flying a version of the Gadsden Flag with 15 stars on it signifying the 15 slave states.

As of 2022[update], the following states offer the option of obtaining a Gadsden flag specialty license plate: Alabama, Arizona,[33] Florida,[34][35] Kansas,[36] Maryland,[37] Missouri, Montana,[38] Oklahoma, South Carolina, Tennessee,[39] Texas, and Virginia.

[45][46] Following Dobbs v. Jackson Women's Health Organization, which struck down Roe v. Wade, abortion rights activists were seen at a Texas rally carrying a version of the flag with the snake in the shape of a human uterus.

[59] In 2014, the flag was used by Jerad and Amanda Miller, the perpetrators of the 2014 Las Vegas shootings who killed two police officers and a citizen.

[61] In March 2013, the Gadsden flag was raised at a vacant armory building in New Rochelle, New York, without permission from city officials.

A federal judge dismissed the case, rejecting the United Veterans' First Amendment argument and ruling that the flagpole in question was city property and thus did not represent private speech.

[63] In 2014, a US Postal Service employee filed a complaint about a coworker repeatedly wearing a hat with a Gadsden Flag motif at work.

Postal service administration dismissed the complaint, but the United States Equal Employment Opportunity Commission reversed the decision and called for a careful investigation.

The EEOC issued a statement clarifying that it did not make any decision that the Gadsden flag was a "racist symbol," or that wearing a depiction of it constituted racial discrimination.

[74] Parodies and pastiches of the Gadsden flag exist; one common design replaces the "Don't tread on me" motto with "No Step on Snek", sometimes paired with a crudely drawn snake.