Difference between revisions of "Part:BBa K1744001:Design"

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===References===
 
===References===
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Mutalik et al., Quantitative estimation of activity and quality for collections of functional genetic elements, Nature methods, 10(4), 2013.
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Vivek K Mutalik, Joao C Guimaraes, Guillaume Cambray, Quynh-Anh Mai1, Marc Juul Christoffersen, Lance Martin, Ayumi Yu, Colin Lam, Cesar Rodriguez, Gaymon Bennett, Jay D Keasling, Drew Endy & Adam P Arkin, Quantitative estimation of activity and quality for collections of functional genetic elements, Nature methods, 10(4), 2013.

Revision as of 17:26, 14 September 2015


P1U8-amilCP-kanR


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1514
    Illegal NheI site found at 1537
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The amilCP chromoprotein synthesis could be improved for the blue color to be visible in single copy faster. It has been of great concern through-out all the project and that is why we have put the strongest combination of promoter and RBS we have found to be well characterised. Still, expression in single copy is not so intense. In our case, amilCP can still be useful as a marker for plasmid background detection during recombineering experiment. Using only the kanamycin marker for recombineering, the blue colonies would represent plasmid transformants and not good recombinants. This use is good for us though, since background is a great concern in recombineering techniques, where screening can be long.



Source

The part was constructed using a kanamycin resistance gene from plasmid pOK12 (available commercially) and amilCP from BBa_K592009. The promoter and RBS P1U8 were synthesised and come from Mutalik et al. (Nature 2013).


References

Vivek K Mutalik, Joao C Guimaraes, Guillaume Cambray, Quynh-Anh Mai1, Marc Juul Christoffersen, Lance Martin, Ayumi Yu, Colin Lam, Cesar Rodriguez, Gaymon Bennett, Jay D Keasling, Drew Endy & Adam P Arkin, Quantitative estimation of activity and quality for collections of functional genetic elements, Nature methods, 10(4), 2013.