| Notes: | Deposited by: J. Justin McCormick The first site was designed for insertion of a marker selectable in mammalian cells, and the second for the sequence to be expressed. The first site permits detection of inserts by alpha complementation. [1] Expression vector containing two multiple cloning sites: the first interrupting the lacZ’ gene, and the second being flanked by a murine metallothionein I promoter and splicing and polyadenylation signals. [1] The plasmid contains the following restriction sites (bp from 0): NotI–0; EcoRI–9, 2894; SacI–1593; StyI–1627; ClaI–1814; XhoI–1836; BglII–1859; SphI–2730; NsiI–2732; PvuI–2774, 4564; HindIII–2945; DrdI–3406, 5277; BsaI–4264. (personal communication) The order of the major features in this plasmid is: NotI – 5′ metallothionein promoter 3′ – ClaI/MCSII’/BglII – poly(A) – SphI – 3’lacZ’ – EcoRI/MCSI/HindIII – 5′ lacZ’ – pMB1 ori – ampR. [1] Restriction digests of the clone give the following sizes (kb): PvuI–3.6, 1.8; EcoRI–2.9, 2.5; HindIII–5.4; BglII–5.4; ClaI–5.4. (ATCC staff) Medium is 1227 LB plus ampicillin. Hosts: E.coli, E.coli HB101, E.coli JM105, E.coli JM109, vertebrate cells. |
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