Composite

Part:BBa_K2332000:Design

Designed by: Paola Handal Marquez   Group: iGEM17_UCL   (2017-09-03)
Revision as of 23:34, 23 October 2017 by Paola handal (Talk | contribs) (→‎References)


GFP-SpyTag (constitutive)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 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 705


Design Notes

To obtain the fusion protein of GFP-SpyTag, we had to remove the stop codon of GFP. Parts were already codon optimized for E. Coli.


Source

SpyTag sequence was obtained from BBa_K1159201 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.