Composite

Part:BBa_K3831032:Design

Designed by: Michaela Dušková   Group: iGEM21_Brno_Czech_Republic   (2021-10-07)


Construct C for testing of Ppho promotor - detecting of phosphate and production of cI repressor


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2301
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 636
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Two genes have been placed under the control of the PPho promoter - reporter protein mScarlet-I to visualize the activity of this promoter and cI repressor (BBa_K3831024) which is a component of our switch system. Repressor cI is able to repress Pgrac-OcI promoter by binding to the sequence of its operator OcI, which was artificially modified. The translation of these coding sequences is then controlled by these RBSs. Their translation rates were predicted using RBS calculator: RBS_R0 + cI repressor (Translation rate = 38 689) RBS_R3 + mScarlet-I (Translation rate = 19 000) Bidirectional terminator BBa_B0014 is located at the 5' and 3' ends of composite part C to prevent the activation of our genes from a native promoter in the chromosome of B. subtilis as well as the activation of B. subtilis genes located downstream from the Pgrac-OcI promoter.


Source

Original sequence

References