Composite

Part:BBa_K1426001:Design

Designed by: Robert H. Newman   Group: iGEM14_Greensboro_Aggies   (2014-10-31)


pLac-RBS-RFP-term-XylR-pXyl-RBS-GFP-2term


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1924
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 781
    Illegal AgeI site found at 893
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3135


Design Notes

Because repression of pLac requires the presence of the lac repressor (LacI), bacterial strains that do not express a functional LacI (such as dh5alpha) exhibit high levels of RFP expression even in the absence of IPTG. Therefore, to achieve inducible expression of the RFP, the part must be expressed in E. coli strains that express a functional form of LacI (e.g., JM109 or BL21).


Source

iGEM parts registry: Parts BBa_J04450 and BBa_K733018

References