Generator

Part:BBa_K524004:Design

Designed by: HO Yuan Heng Trevor   Group: iGEM11_HKUST-Hong_Kong   (2011-10-03)

pir gene


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 438
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Homologous sites in primers designed by researchers to clone the pir gene into the E. coli BW25141 genome were used to clone out and standardize the pir gene. This is to avoid truncation of pir gene, and therefore junk sequences of 154 base pairs can be found downstream of the pir CDS.

Source

Cloned out and standardized from the genome of strain BW25141 (courtesy of The Coli Genetic Stock Center).

References

F Wu, I Goldberg, and M Filutowicz.(1992).Roles of a 106-bp origin enhancer and Escherichia coli DnaA protein in replication of plasmid R6K, Nucleic Acids Res. 1992 February 25; 20(4): 811–817.

F Wu, I Goldberg, and M Filutowicz.(1994).Binding of DnaA protein to a replication enhancer counteracts the inhibition of plasmid R6K gamma origin replication mediated by elevated levels of R6K pi protein, J Bacteriol. 1994 November; 176(22): 6795–6801.

M Filutowicz, G Davis, A Greener, and D R Helinski.(1985).Autorepressor properties of the pi-initiation protein encoded by plasmid R6K, Nucleic Acids Res. 1985 January 11; 13(1): 103–114.

M Filutowicz, M J McEachern, and D R Helinski.(1986).Positive and negative roles of an initiator protein at an origin of replication, Proc Natl Acad Sci U S A. 1986 December; 83(24): 9645–9649.