Composite

Part:BBa_K1744002:Design

Designed by: Frederic Grenier   Group: iGEM15_Sherbrooke   (2015-09-14)
Revision as of 06:56, 15 September 2015 by Fred us (Talk | contribs) (→‎Reference)

Design notes

The killswitch is induced by arabinose. Since arabinose induction is highly dependent on cell metabolism, make sure that the cells will keep their metabolism toward glucose by adding glucose to the medium. To induce the toxin, both in genomic context or in plasmidic context, use arabinose.


Source

The part was constructed using araC and PBAD of pBAD30 (commercially available), a synthetic RBS from Mutalik et al. (2013), the toxin vcrx028 from the conjugative plasmid pVCR94, P1U8, a strong constitutive promoter and a strong RBS from Mutalik et al. (Nature 2013), amilCP, a chromoprotein isolated from Acropora millepora that was BioBrick adapted in E.coli by a previous IGEM team (see BBa_K592009) and KanR (Aph(3’)-I (aminoglycoside


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.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1227
    Illegal NheI site found at 1630
    Illegal NheI site found at 1653
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1166
    Illegal BamHI site found at 1613
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1001
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 983