Restriction digests of the clone give the following sizes (kb): PvuII--4.0, 1.5; PstI--3.1, 2.4; BamHI--5.5; SmaI--5.5; XbaI--5.5. This is an expression vector capable of producing a B-gal: insert fusion protein. It can also be used as a probe. 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 terminated before APH(3')-I. 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). 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. |
