pE5LVP0

Contains DNA corresponding to nt 260 through 2004 of encephalomyocarditis virus RNA, including the 5′ non-coding region & coding regions for L, 1A, and IB. Permits increased expression for in vitro transcription translation systems by providing a translational enhancer downstream of the T7 promoter. Digestion of the plasmid with XbaI plus BalI generates a fragment (approximately 4 kb) that has no EMC coding regions, but retains the translational enhancer. This can be used for cloning. If a potential insert contains transcription terminators, then the BalI site can be used for cloning. For in vitro transcription, the plasmid should be linearized with XbaI. The order of the major features in this plasmid is:T7 promoter – 5′ non-coding region of EMC – BalI – partial coding regions of EMC – XbaI – MCS – pSPT18. Restriction digests of the clone give the following sizes (kb): EcoRI–4.8; HindIII–3.28, 1.54; XbaI–4.8. (personal communication) Medium is 1227 LB plus ampicillin. Hosts: E.coli C600, E.coli, E.coli HB101.