Composite

Part:BBa_K3725120

Designed by: Monica Cho, Sahana Narayanan, Manasvi Gupta, Janet Standeven   Group: iGEM21_Lambert_GA   (2021-09-30)
Revision as of 19:22, 17 December 2021 by Manasvigupta (Talk | contribs) (EXPERIENCE)


PhoB Mutated Consensus Promoter with GFP Reporter

DESIGN

The promoter BBa_K3725130 was designed by replacing parts of BBa_K116401 with the PhoB consensus sequence. A consensus sequence is a calculated order of the most repeated nucleotides found at each position in an alignment of multiple other sequences that have a similar function. By replacing the downstream Pho Regulon with GFP, the part was expected to express fluorescence in the presence of low levels of extracellular phosphate.

EXPERIENCE

Utilizing the modified phosphate range and characterization protocol (See: Modified Protocol), we characterized the BBa_K3725120 phosphate sensor. However, after overnight incubation, we noticed that the cells produced no fluorescence. Despite resuspending the cells in MOPS media, which contains relatively less phosphate than LB, the part did not exhibit greater fluorescence. Since we confirmed the cloning of the part with successful sequencing results, we concluded that the part was not efficient as a phosphate sensor and discontinued characterization.



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 BsaI.rc site found at 1148


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