Reporter

Part:BBa_K2333441:Design

Designed by: Ethan M Jones   Group: iGEM17_William_and_Mary   (2017-10-27)
Revision as of 23:53, 29 October 2017 by Chli (Talk | contribs)


Copper sensor with Protein Degradation Tag E


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1229
    Illegal AgeI site found at 1341
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Parts of this had to be ordered as a G-block and was assembled via Gibson Assembly.


Source

The CueR and PcopA sequences were taken from E. coli. CueR was codon optimized.

Pdt E was originally generated by mutagenesis from the endogenous Lon degraded tags from the bacteria Mycoplasma florum by Collins et al. 2014 "Tunable Protein Degradation in Bacteria". Pdt E corresponds to Collins et al.'s tag pdt#3d. To create pdt E, the amino acid sequence was taken from Collins et al. and was codon optimized for E. coli, then synthesized by IDT.

UNS sequences are from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly.


References