Restriction digests of the clone give the following sizes (kb): BamHI--5.45; HindIII--5.45; EcoRI--5.45; PvuII--4.0, 1.5; PstI--3.1, 2.35. This is an expression vector capable of producing a beta-galactosidase: insert: APH(3')-I fusion protein. The start codon of the modified APH(3')-I is followed by the eleventh codon (serine) of APH(3')-I. Codons 2-10 have been deleted. The upstream translation of LacZ is out of frame with APH(3')-I. Sequences from the TAA stop codon of APH(3')-I to the immediate downstream PvuII site encode a portion of Tn601. Sequences from the PvuII site to the PstI site are derived from Tn5. The vector contains pBR322 (EcoRI-SphI), a TAC promoter, the first 8 amino-terminal codons of beta-galactosidase, and a gene encoding a modified aminoglycoside phosphotransferase (3')-I (APH(3')-I). |
