User:Jeffrey Kim/Notebook/Halogenated natural products from uncultured bacteria/2008/12/09

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FAD halogenase

  • Started cloning the ZA41 ZB118 pathway together. The orientation looks appropriate so an AvrII digest of the fragments, dephosphorylation of ZA41, and double selection on Amp and Kan should select for the appropriate recombinant. Digests and ligation performed as per mfg. and plated controls of vector alone look fine (no background after CIP dephosphorylation). Restriction maps of the resulting clones does not look appropriate. Diagnostic digest of ZB118 shows that it may be missing a site.