Composite

Part:BBa_K4676006:Design

Designed by: Nicholas H. Perez   Group: iGEM23_Hopkins   (2023-10-11)


mCherry Transcriptional Unit


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 444
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 444
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 444
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 444
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

When choosing the promoter, it was important to choose a promoter that would work in T7 strain E. coli; given that T7 Shuffle strain was specifically selected as a chassis, in order to boost disulfide bond formation between the H-Fibroin and the L-Fibroin silk proteins.

The mCherry was chosen because T7 Shuffle E. coli are resistant to the same antibiotic, Spectinomycin, used to plate the level 2 Golden Gate plasmid backbone (pJUMP49-2A(sfGFP)) used in this project. The red fluorescence thus allows researchers to pick only the bacterial colonies that have taken up the level 2 assembly, ensuring more efficient protein purification.


Source

See pages for each of the individual parts.

References