The main difference between the settlement of Jamestown and of the Plymouth was "economic motives and religious freedom".
The settlers of Jamestown colony had economic reasons for their settlement in Jamestown, whereas immigrants moved in the colony of the Plymouth to have religious freedom. The colony of Jamestown was founded to expand the trade and to develop a market for English manufactured goods. On the other hand, settlers of the Plymouth colony were seeking freedom of worship. Settlers here even signed a document called Mayflower Compact to begin a colonial self-government.