Difference between revisions of "Part:BBa K1744002:Design"
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| − | The part was constructed using araC and PBAD of pBAD30 (commercially available), a RBS selected through rational design, the toxin vcrx028 from the conjugative plasmid pVCR94, P1U8 that are 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 ( | + | The part was constructed using araC and PBAD of pBAD30 (commercially available), a RBS selected through rational design, the toxin vcrx028 from the conjugative plasmid pVCR94, P1U8 that are 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 (aminoglycoside3’-phosphotransferase)) from pOK12 (it is a truncated version). In practice, this part was built from two previously submitted parts of our team: BBa_K1744000 and BBa_K1744001. |
===References=== | ===References=== | ||
Revision as of 07:01, 15 September 2015
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.
Sources
The part was constructed using araC and PBAD of pBAD30 (commercially available), a RBS selected through rational design, the toxin vcrx028 from the conjugative plasmid pVCR94, P1U8 that are 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 (aminoglycoside3’-phosphotransferase)) from pOK12 (it is a truncated version). In practice, this part was built from two previously submitted parts of our team: BBa_K1744000 and BBa_K1744001.
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1227
Illegal NheI site found at 1630
Illegal NheI site found at 1653 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1166
Illegal BamHI site found at 1613 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1001
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 983