Composite

Part:BBa_K2332041:Design

Designed by: Paola Handal   Group: iGEM17_UCL   (2017-10-20)


Blue light inducible expression of GFP-SpyCatcher (Pblind GFP-SpyCatcher)


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 741


Design Notes

We removed the stop codon in GFP and placed the SpyCatcher sequence after a linker sequence and included a HisTag for protein purification. Pblind promoter was designed by Jayaraman P. et al. (2016)

Source

SpyCatcher sequence was obtained from BBa_K1159200 and GFP from: BBa_E0040. Pblind promoter was designed by Jayaraman P. et al. (2016). The DNA sequence was synthesised by Integrated DNA Technologies (IDT)

References

1. Jayaraman P, Devarajan K, Chua T, Zhang H, Gunawan E, Poh C. Blue light-mediated transcriptional activation and repression of gene expression in bacteria. Nucleic Acids Research. 2016;44(14):6994-7005.

2. Zakeri B, Fierer J, Celik E, Chittock E, Schwarz-Linek U, Moy V et al. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proceedings of the National Academy of Sciences. 2012;109(12):E690-E697.