Composite

Part:BBa_M50470:Design

Designed by: Oluremi Akindele   Group: Stanford BIOE44 - S11   (2018-10-18)


Cold Shock Sensor w/ aeBlue+3X-FLAG tag Pigment Reporter


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
    COMPATIBLE WITH RFC[1000]


Design Notes

We added an aeBlue chromoprotein sequence with an RBS taken from BBa_K1033929 downstream of our cspA promoter (Bba_K328000) such that the cspA promoter controls transcription of the aeBlue chromoprotein. Behind the chromoprotein sequence is a 3X-FLAG epitope tag (Bba_K823034) that we can use for Western blotting and other protein separation experiments.

Source

aeBlue chromoprotein: BBa_K1033929 cspA promoter: Bba_K328000 3X-FLAG tag: Bba_K823034

References

https://parts.igem.org/Part:BBa_K328000 https://parts.igem.org/Part:BBa_K1033929 https://parts.igem.org/Part:BBa_K823034 Role of Escherichia coli cspA promoter sequences and adaptation of translational apparatus in the cold shock response: https://www.ncbi.nlm.nih.gov/pubmed/9393453 Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology: https://www.ncbi.nlm.nih.gov/pubmed/29760772 Agashe, V. R., & Hartl, F. (2000). Roles of molecular chaperones in cytoplasmic protein folding. Seminars in Cell & Developmental Biology, 11(1), 15-25.