Difference between revisions of "Part:BBa K2333434:Design"

 
 
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===Design Notes===
 
===Design Notes===
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This composite part was designed with the LacI repressor under the constitutive promoter J23105 and mf-Lon under the control of the PLlac 0-1 promoter. The mf-Lon gene was modified via codon-optimization for iGEM use and a double terminator was added.
  
  
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===Source===
 
===Source===
  
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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.
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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".
  
 
===References===
 
===References===
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[1] Cameron DE, Collins JJ. Tunable protein degradation in bacteria. Nature Biotechnology. 2014;32(12):1276–1281.
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[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.

Latest revision as of 22:35, 29 October 2017


pLac0-1 mf-Lon


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 47
    Illegal NheI site found at 70
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 3242
    Illegal AgeI site found at 3326
    Illegal AgeI site found at 3532
    Illegal AgeI site found at 3557
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This composite part was designed with the LacI repressor under the constitutive promoter J23105 and mf-Lon under the control of the PLlac 0-1 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.