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The whole nucleotide sequence of pHY300PLK (Ishiwa and Shibahara 1985a) has been determined, pHY300PLK consists of 4870 bp and contains an RNA primer for ori-177 and three open reading frames corresponding to TcR, ApR and Rep-α1. Twenty-three unique restriction endonuclease recognition sites were found. We have confirmed that ORF α1 of pHY300PLK is the plasmid replication gene. A rescue phenomenon has been found with respect to multiple infection by pHY300.2PLK (rep-) and pUB110. A series of experiments indicated that the oligomer form, pHY300.2PLK, was rescued by the monomeric form, pUB110. Additionally, some useful information for using pHY300PLK as a shuttle vector in molecular cloning studies is presented. (Key words: shuttle vector; E, coli; B, subtilis; tetracycline; ampicillin; kanamycin; rescue; MIC).

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New shuttle vectors for Escherichia coli and Bacillus subtilis. IV The nucleotide sequence of pHY300PLK and some properties in relation to transformation.