I purchased a plasmid from the myTXTL Toolbox 2.0 Plasmid Collection. Is it possible to use the plasmid from the original tube directly for in vitro protein production?

No. All our Toolbox 2.0 plasmids (except the positive control plasmid P70a-deGFP that comes with the myTXTL kit are meant for plasmid amplification in E. coli only. The degree of purity is NOT sufficient for efficient in vitro production. Please refer to the current myTXTL handbook for recommendations on preparation of plasmid templates for myTXTL reactions.