Composite

Part:BBa_K1156000:Design

Designed by: Haylee Bachman   Group: iGEM13_GeorgiaTech   (2013-09-17)


RBS_T7_Pet(mCherry)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1826
    Illegal AgeI site found at 1700
    Illegal AgeI site found at 1793
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To comply with BioBrick standard 10, Phe-76 was re-optimized to remove a EcoRI site within the part. A T7 promoter with a lacI operator was added with the RBS (AGGA) to the PET_mCherry. A standard assembly 10 prefix and suffix was then added.


Source

A sequence optimized for E. coli for a PET beta barrel auto-transporter expressing mCherry was obtained from Sevastynovich et al."A generalised module for the selective extracellular accumulation of recombinant proteins." Microbial Cell Factories. 2012. The sequence was assembled from synthesized 500bp blocks through IDTdna "Gblock" synthesis.

References

Sevastynovich et al."A generalised module for the selective extracellular accumulation of recombinant proteins." Microbial Cell Factories. 2012.