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. 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. 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. 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. 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. |
