Restriction digests of the clone give the following sizes (kb): BglII--2.9, 2.6; EcoRI--5.6; HindIII--2.9, 1.3, 1.1, 0.41; BamHI--5.6. The ADE2 segment contains HindIII sites approximately 1.17 kb and 2.36 kb from the 5' BglII site. A YR-type shuttle expression vector containing promoters for in vitro RNA synthesis and priming sites useful for sequencing. Contains readily purifiable elements for construction experiments. In E. coli, permits visual detection of recombinants and production of ssDNA. Encodes the lacZalpha (lacZ') peptide. The naturally occurring BglII site within ADE2 was destroyed by replacing the A at nt 1243 of the reported sequence with a G. A BglII linker was added to the 5' end, and the naturally occurring BglII site at the 3' end (nt 2512) was used to create a BglII fragment containing the complete coding sequence. pTZ19B was derived from pTZ19R by adding a BglII linker at coordinate 416. Constructed by inserting a 2.5 kb fragment (Ars+ Ade2+) into the BglII site of pTZ19B. The order of the major features in this plasmid is: lacZ'/MCS - T7 promoter - ADE2 - pMB1 ori - ampR - f1 ori. |
