Restriction digests of the clone give the following sizes (kb): PvuI--3.4, 1.8; EcoRI--2.6, 2.6; BglII--5.2; ClaI--4.1, 1.1. The plasmid contains the following restriction sites (bp from 0): NotI--0; EcoRI--9, 2640; ClaI--425, 1560; DrdI--1398, 3152, 5021; BglII--1605; SphI--2476; NsiI--2478; PvuI--2522, 4310; HindIII--2691; BsaI--4010. Expression vector containing two multiple cloning sites: the first being within the lacZ' gene, and the second being flanked by an RSV enhancer-MMTV LTR promoter at one end and splicing and polyadenylation signals at the other. 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/EcoRI - 5' RSV enhancer - MMTV LTR 3' - ClaI/MCSII'/BglII - splicing - polyadenylation - NsiI - 3' lacZ' - EcoRI/MCSI/HindIII - 5' lacZ' - pMB1 ori - ampR. |
