Composite

Part:BBa_K2333402:Design

Designed by: Ethan M Jones   Group: iGEM17_William_and_Mary   (2017-06-16)
Revision as of 19:43, 16 June 2017 by CiCi Zheng (Talk | contribs) (→‎Source)


Cloning ready protein degradation tag B (medium-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 tag, which enables it to be appended to an arbitrary protein in a given circuit, without changing the underlying architecture.


Source

The tag pdt #3a is 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". We synthesized the tag by IDT and cloned using gibson assembly.

B0015 comes from the registry, and the UNS sequence comes 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.

UNS 2 Sequence is a unique oligonucleotide sequence introduced by William and Mary iGEM 2016 . See BBa_K2066018 and BBa_K2066019.

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] Part:BBa_K2066018. Part:BBa K2066018. [accessed 2017 Jun 16]. https://parts.igem.org/Part:BBa_K2066018

[4] Part:BBa_K2066018. Part:BBa K2066019. [accessed 2017 Jun 16]. https://parts.igem.org/Part:BBa_K2066019