Part:BBa_M50059:Design

Blackteria: Genetic sequence for the production of melanin

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 239
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 614
  

Design Notes

A T5 promoter is easily inducible and therefore an effective promoter to test the melanin sequence. The strong RBS makes sure that the ribosome binds. The double terminating sequence makes sure transcription terminates. The original DNA sequence comes from part Bba_K274001 which obtained sequence from Rhizobium etli CFN42. However, we optimized the sequence for E. coli so that it will work better. The optimization improves the compatibility of the plasmid by changing the base pairs without changing the codon produced.

Source

Rhizobium etli CFN42, unaltered gene sequence obtained from: BBa_K274001

References

http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2672.2006.03013.x/full , make-up of the media including necessary Cu and L-tyrosine precursor. http://2009.igem.org/Team:Tokyo_Tech/BlackenedEcoli#Materials_and_Methods http://partsregistry.org/Part:BBa_K274001 "Team:Tokyo Tech/Blackened E. coli." Team:Tokyo Tech/Blackened E. coli. IGEM.org, n.d. Web. 9 May 2017. Lagunas-Muñoz, V.h., et al. 2006. "Optimum Melanin Production Using Recombinant Escherichia Coli." Journal of Applied Microbiology 101(5): 1002-008. Web. “What’s that Stuff?” 1998. Chemical and Engineering News 76(46): 51-56. Web. "Part:BBa_R0010:Experience." Registry of Standard Biological Parts. IGEM, n.d. Web. 9 May 2017. “MOPS EZ Rich Defined Medium.” Teknova: Science Matters. n.d. Web. 10 June 2017. Retrieved from: http://www.teknova.com/EZ-RICH-DEFINED-MEDIUM-KIT-p/m2105.htm “Endy: E. coli Wetern Blot.” 2007. OpenWetWare. Retrieved from: http://www.openwetware.org/wiki/Endy:E._coli_Western_Blot