Composite

Part:BBa_K2333401:Design

Designed by: Ethan M Jones   Group: iGEM17_William_and_Mary   (2017-06-14)
Revision as of 17:21, 28 October 2017 by Ethan801 (Talk | contribs) (→‎References)


Cloning ready protein degradation tag A (strong) with double terminator


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 site found at 41
    Illegal BsaI.rc site found at 263


Design Notes

This part was designed to include both a double stop codon and double terminator after the pdt, which enables it to be appended to an arbitrary protein in a given circuit, without changing the underlying architecture.

Source

Pdt A 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 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

[1] Torella JP, Boehm CR, Lienert F, Chen J-H, Way JC, Silver PA. Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic Acids Research. 2013;42(1):681–689.

[2] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.

[3] BBa_K2066018.Bba_ K2066018.

[4] BBa_K2066018. BBa K2066019.