SmaI, HincII and EcoRI sites appear in both the polylinker and the aph cassette. Restriction digests of the clone give the following sizes (kb): HindIII--4.65; HincII--4.65; PstI--3.0, 1.7. An Escherichia coli/eukaryotic cell line shuttle vector containing lacZ and multiple cloning sites of pUC18 and the aph (neoR) gene under the control of herpes simplex virus tk promoter. A PvuII (position 628) of pUC18 was changed to BglII. The aph (aminoglycoside phosphotransferase) gene under the control of herpes simplex virus tk promoter was inserted at the BglII site. Aph is transcribed in the same direction as lacZ. |
