Difference between revisions of "Part:BBa K2066111:Design"
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===Design Notes===
 
===Design Notes===
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UNS 6.1 was used as a spacer. This part was created to reduce metabolic strain from maintaining two separate plasmids for the reporter and repressor.
 
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pLacO1 controlling sfGFP expression and constitutively expressed LacI were placed onto the same Biobrick backbone flanked by UNS sequences to allow for easy cloning.  This reduces the metabolic stress on the cells being transformed.
  
 
===Source===
 
===Source===
  
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sfGFP and RBS from Lou et al. 2012, “Ribozyme-based insulator parts buffer synthetic circuits from genetic context” UNS sequences from Torella et al. 2013 (“Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly”
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The sfGFP flourescent reporter and pLacO1 design is inspired by C. Lou, B. Stanton, Y.-J. Chen, B. Munsky, C. A. Voigt, Ribozyme-based insulator parts buffer synthetic circuits from genetic context. Nat. Biotechnol. 30, 1137 (2012). doi:10.1038/nbt.2401 pmid:23034349. The UNS sequences at the ends of the insert are derived from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013).
  
 
===References===
 
===References===

Latest revision as of 06:23, 29 October 2016


pLacO1 sfGFP + LacI (weak)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1087
    Illegal NheI site found at 1110
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 224


Design Notes

pLacO1 controlling sfGFP expression and constitutively expressed LacI were placed onto the same Biobrick backbone flanked by UNS sequences to allow for easy cloning. This reduces the metabolic stress on the cells being transformed.

Source

The sfGFP flourescent reporter and pLacO1 design is inspired by C. Lou, B. Stanton, Y.-J. Chen, B. Munsky, C. A. Voigt, Ribozyme-based insulator parts buffer synthetic circuits from genetic context. Nat. Biotechnol. 30, 1137 (2012). doi:10.1038/nbt.2401 pmid:23034349. The UNS sequences at the ends of the insert are derived from Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013).

References