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WASHINGTON, D.C.—Jeffrey Gladney teaches the Old Testament at Northeast Mississippi Community College in Tupelo, Miss.

To celebrate the holiday season, he brought his wife and children to the new Museum of the Bible in the nation’s capital—a 430,000-square-foot interactive experience that tells the history and impact of the Old and New Testaments.  

“It’s like a refresher course to me,” he says. “It helps us have a visual image of what happened during the life of Jesus.”

The $500 million museum opened with much fanfare last month just two blocks from the National Mall and three blocks from the U.S. Capitol. The brainchild of Hobby Lobby craft chain owners Steve and Jackie Green, it is one of the largest museums about the Bible in the world.

It has more than 3,000 artifacts, including fragments of the Dead Sea Scrolls, a first edition of the King James Bible, and a page from the Gutenberg Bible, the first major book to use moveable metal type in Europe. About 2,000 of those items are on loan from more than 41 other collections and institutions, including the Israel Antiquities Authority, which donated a one-ton stone from the Western Wall.

Museum executives and employees say their mission is to educate, not evangelize—and to welcome visitors of all faiths and cultures.

“You don’t have to love the Bible to think it’s an incredible story,” says Danielle Smith, social manager at the museum. “There’s an understanding that the Bible has impacted the world. You see it in literature, fashion, architecture. This is a book that should be known.”

On a recent morning, visitors cued up to walk through the entrance hall of the six-floor building, which is lined by columns made from Jerusalem stone.

“What…