Restriction digests of the clone give the following sizes (kb): PvuII--3.0, 1.5; HincII--3.0, 1.5; EcoRI--4.25, 0.3. A plasmid shuttle vector for investigating promoter and/or enhancer elements. Has a polylinker upstream of reporter CAT gene, SP6 and T7 promoters for in vitro RNA synthesis and low basal CAT expression in eukaryotic cells. The sequence at the junction of the multiple cloning site and CAT is GAATTCGAGC and at the junction of SV40 and SP6 is GGATCAGCTT. The polylinker is in opposite orientation in pGCAT-A (ATCC 37640) and pGCAT-C (ATCC 37641). BamHI/HindIII fragment of pSB1 (CAT) was cloned into the HindIII site of pGEM-4. 2 hrs after ligation began the remaining BamHI and HindIII sites were filled in. Contains SV40 polyadenylation site and donor and acceptor sites of SV40 small-t intron. |
