Coding

Part:BBa_K1323003

Designed by: Julia Manalil   Group: iGEM14_Waterloo   (2014-09-04)

oriT-Nes: expression sequence for Staphylococcus conjugation

oriT-Nes is a 2.2 kb region that includes an origin of transfer (oriT) and a gene that encodes the oriT-nicking protein (Nes). This sequence originates from the staphylococcal plasmid, pGO1 (Caryl & O’Neill, 2008). When a plasmid in a gram-positive organism undergoes conjugation a multiprotein complex binds at the oriT to form a relaxosome. The relaxosome is recognized by DNA relaxases that then cut the DNA. This cut is necessary for the transfer of the DNA from the cell (Grohmann et al, 2003). Our team obtained the pGO1 plasmid from Dr. Alex O’Neill’s lab at the University of Leeds. We amplified the oriT-Nes region out of pGO1 by PCR.


Usage and Biology

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

Caryl, J. A., & O’Neill, A. J. (2009). Complete nucleotide sequence of pGO1, the prototype conjugative plasmid from the Staphylococci. Plasmid, 62(1), 35–8.

Grohmann, E., Muth, G., Espinosa, M., Grohmann, E., & Espinosa, M. (2003). Conjugative plasmid transfer in gram-positive bacteria. Microbiology and Molecular Biology Reviews, 67(2), 277–301.


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