Composite

Part:BBa_I746666:Design

Designed by: Yue Miao   Group: iGEM07_Cambridge   (2007-10-23)

Pspac-hy with GFP


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 728


Design Notes

Note that there is a G->T point mutation at the -1 site, changing the promoter to a stronger one - hence the name Pspac-hy, hy meaning hyper. This promoter drives transcription at rates of about ten times that of the wild type (Pspac) promoter.

This change is documented in the reference article cited below.

Source

Comes from the pPL82 vector, which is intended as a B. subtilis integrational vector. It is placed in front of the MCS in the plasmid, as it would drive the expression of the gene cloned into the MCS both in B. subtilis and E. coli.

References

  • Yansura DG, Henner DJ. 1984. Use of the Escherichia coli lac repressor and operator to control gene expression in Bacillus subtilis. Proc Natl Acad Sci U S A. 1984 Jan;81(2):439-43. ([http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6420789&dopt=AbstractPlus])
  • Quisel JD, Burkholder WF, Grossman AD. In vivo effects of sporulation kinases on mutant Spo0A proteins in Bacillus subtilis. J Bacteriol. 2001 Nov;183(22):6573-8. ([http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=11673427])