Difference between revisions of "Part:BBa K2333435:Design"
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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.
 
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.
  
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The sequence for the mf-Lon gene orthogonal to the endogenous machinery in E. coli was obtained from the paper by Collins et al. 2014 "Tunable Protein Degradation in Bacteria."
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The sequence for the mf-Lon gene codon-optimized for E. coli expression was obtained from the paper by Collins et al. 2014 "Tunable Protein Degradation in Bacteria."
  
  

Revision as of 22:34, 29 October 2017


pBad mf-Lon


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1245
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1184
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1019
    Illegal AgeI site found at 3102
    Illegal AgeI site found at 3186
    Illegal AgeI site found at 3392
    Illegal AgeI site found at 3417
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1001


Design Notes

This part was designed with mf-Lon under the control of the inducible pBad promoter. The mf-Lon gene was modified via codon-optimization for iGEM use and a double terminator was added.


Source

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.

The sequence for the mf-Lon gene codon-optimized for E. coli expression was obtained from the paper by Collins et al. 2014 "Tunable Protein Degradation in Bacteria."


References

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

[2] 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.