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Next-generation sequencing of pIJ101-based Streptomyces-E. coli shuttle vectors

Katelyn V. BrowniD, S. Eric NyboiD

Abstract

The sequences of two commonly used pIJ101-based Streptomyces-E. coli shuttle vectors, pEM4 and pUWL201, were determined using next-generation sequencing and mapped. pEM4 was found to be 8.3 kbp long, containing genes encoding {beta}-lactamase, thiostrepton resistance, the lacZ{square} fragment, an E. coli pUC19 origin of replication, and the Streptomyces pIJ101 origin of replication. pUWL201 was found to be 6.78 kbp long, containing genes encoding {beta}-lactamase, thiostrepton resistance, the lacZ{square} fragment, an E. coli pUC19 origin of replication, and the Streptomyces pIJ101 origin of replication. Interestingly, the sequences for both pEM4 and pUWL201 exceed their previously reported size by 1.1 kbp and 0.4 kbp, respectively. This report updates the literature with the corrected sequences for these two popular vectors, ensuring their compatibility with modern synthetic biology cloning methodologies.

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