Composite

Part:BBa_M50499:Design

Designed by: Teaghan Cowles, Rishabh Kapoor, Alex Bradfield, Yash Pershad   Group: Stanford BIOE44 - S11   (2018-10-21)
Revision as of 23:53, 11 December 2018 by TRAY (Talk | contribs)


Catabolite Activator Protein (CAP) Repressed Promoter

We placed the constitutive promoter (BBa_S05450, iGEM) immediately upstream from a catabolite activator protein (CAP) binding site (BBa_M36547, iGEM). When CAP binds to the binding site, expression will be repressed.


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1059
    Illegal PstI site found at 688
    Illegal PstI site found at 849
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 144
    Illegal NheI site found at 167
    Illegal PstI site found at 688
    Illegal PstI site found at 849
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1059
    Illegal PstI site found at 688
    Illegal PstI site found at 849
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1059
    Illegal PstI site found at 688
    Illegal PstI site found at 849
  • 1000
    COMPATIBLE WITH RFC[1000]

This construct is an adaption of the natural lac operon, which is positively regulated by CAP, since the CAP binding site is naturally upstream of the RNA polymerase binding site. In this construct, we put the CAP binding site downstream in order to repress transcription due to steric hindrance when CAP binds.

Design Notes

This construct is an adaption of the natural lac operon, which is positively regulated by CAP, since the CAP binding site is naturally upstream of the RNA polymerase binding site. In this construct, we put the CAP binding site downstream in order to repress transcription due to steric hindrance when CAP binds.

Since CAP binding is inversely correlated to glucose concentrations (and positively correlated with cyclic AMP (cAMP) levels), this promoter exhibits glucose-inducibility and cAMP-repressibility. We employed this construct as a potential DNA-based glucose biosensor in E coli.

Source

IGEM part: BBa_S05450 = constitutive promoter IGEM part: BBa_M36547 = CRP binding site

Paper: www.ncbi.nlm.nih.gov/pmc/articles/PMC338411/

References