Reporter

Part:BBa_K216015:Design

Designed by: Edinburgh iGEM 2009   Group: iGEM09_Edinburgh   (2009-10-16)


PyeaR with firefly luciferase


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 SapI.rc site found at 932


Design Notes

No special considerations. Note that sequencing shows an unexpected insertion of 6 bases, ACCACC, after the scar between the ribosome binding site and the ATG of the luciferase coding sequence; that is, the sequence reads ACTAGACCACCATG rather than ACTAGATG. If this is correct, these 6 bases must be present in luciferase BioBrick BBa_I712019, or else were introduced by a random event during construction. This makes the RBS 12 bases from the ATG, somewhat more than the optimum, but with a strong RBS (like J15001) should still give about 25% of the expression level expected for optimum spacing in E. coli (Vellanoweth & Rabinowitz, 1992, Molecular Microbiology 6, 1105-1114).


Source

Made by joining PyeaR BioBrick BBa_K216005 to a PCR fusion product of strong synthetic ribosome binding site J15001 and firefly luciferase BioBrick BBa_I712019.

References