Generator

Part:BBa_K415205:Design

Designed by: Grant Robinson   Group: iGEM10_MIT   (2010-08-20)


J23117 : B0031 : cymR Repressor Protein [Weak cymR Generator]


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 420
    Illegal XhoI site found at 201
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 103


Design Notes

Note that a weak promoter was chosen in order to follow the construction design of Choi, et al. in the 2010 paper, "A novel, versatile, and tightly regulated expression system for E. Coli strains." Previous work by Choi and others in M. Extorquens indicated that overexpression of cymR was potentially toxic to bacterial cells; although data concerning this toxicity is unavailable, the researchers managed to develop a tightly expressed cymR switch by placing cymR under the control of a weak, constitutive promoter for kanamycin resistance. As our team was unable to clarify the origin of this sequence, we instead selected a weak, constitutive promoter from the Anderson library.


Source

This part consists of BioBricks BBa_J23117 and BBa_K415203.

References