Composite

Part:BBa_K4779001:Design

Designed by: Ziqi Mi   Group: iGEM23_Nanjing-BioXstem   (2023-09-26)


CMPG Pro:A yeast copper-induced reporter pathway with prm1 pro promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 279
    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 55
    Illegal BamHI site found at 510
    Illegal XhoI site found at 8
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 279
    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 279
    Illegal XbaI site found at 517
    Illegal SpeI site found at 343
    Illegal SpeI site found at 523
    Illegal PstI site found at 49
    Illegal NgoMIV site found at 36
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2181
    Illegal SapI site found at 529


Design Notes

When designing the sequence in detail, one of the key considerations is how to convert copper ion signals into information molecule signals more efficiently and sensitively. In this round of DBTL, we selected a more sensitive pprm1 Pro promoter to replace pprm1, thereby enhancing the expression intensity of GFP under the information molecule promoter. pprm1 Pro (BBa_K3384313) is a more sensitive information molecule-responsive promoter, containing three ×PREs in the same direction as the promoter, providing better sensitivity to information molecules.


Source

CMPG pro is a new composite part (BBa_K4779001). We obtained the gene sequences of the promoter of Cup1 (BBa_K945002), MFα2 (BBa_K110005), the promoter of prm1 pro (BBa_K3384313), green fluorescent protein (GFP) (BBa_K3112009), and the terminator CYC1 (BBa_K4278703) from IGEM, and sent them to GENEWIZ (Suzhou genewiz Biotechnology Co., Ltd.) for synthesis.

References