1887

Abstract

The indigenous plasmid pIJ101 is the parent of many cloning vectors used in . One early pIJ101 derivative, pIJ702, has been particularly widely used. pIJ702 lacks : and accumulates single-stranded DNA (ssDNA). The 1.2 kb I-I :cop/ and 0.7 kb I-I regions were isolated from pIJ101 and cloned into pIJ702 at the I site in both orientations. No ssDNA was detected in constructs containing present in its correct orientation with respect to the basic replicon, with or without . Constructs which contained in the reverse orientation did accumulate ssDNA. Thus, is only active as the site for second-strand synthesis in its natural orientation. Furthermore, inserted in either orientation into the structurally unstable pIJ702-pUC8 shuttle vectors prevented them from rearranging in . The function was defined to a 0.53 kb I-II fragment and the probable site for second-strand initiation () was identified.

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1993-04-01
2021-05-07
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