pCAP03

Use

The pCAP03 plasmid has been successfully used to create capture vectors that can be used for TAR cloning of biosynthetic gene clusters. 'Capture arms' for a target BGC can be cloned within the restriction enzymes sites, and the plasmid contains elements that allow transfer between E. coli, actinobacteria and yeast cells.

Features

• Apramycin resistance cassette (acc(3)IV)
• Neomycin and kanamycin resistance
• ΦC31 integrative site & integrase
• pUC ori for maintenance in E. coli
• Yeast elements: autonomous replication sequence ARSH4, centromere CEN6
• Selection markers TRP1 and URA3

History

The plasmid was developed by Bradley Moore's lab as an upgraded version of pCAP01¹.

Map

Sequence links

See AddGene link (here) to access the sequence.

References

The paper reporting pCAP03 is here

1. Yamanaka. K., Reynolds. K.A., Kersten. R.D., Ryan. K.S., Gonzalez. D.J., Nizet. V., Dorrestein. P.C., Moore. B.S. (2014) Direct cloning and refactoring of a silent lipopeptide biosynthetic gene cluster yields the antibiotic taromycin A. Proceedings of the National Academy of Sciences of the United States of America 111(5):1957-62. doi: 10.1073/pnas.1319584111