Reporter

Part:BBa_K274002:Design

Designed by: Shuna Gould   Group: iGEM09_Cambridge   (2009-10-08)
Revision as of 15:29, 19 October 2009 by Shuna (Talk | contribs) (→‎Source)

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Vio operon


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1029
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 5602
    Illegal BamHI site found at 4289
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3814
    Illegal AgeI site found at 4010
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 6552
    Illegal SapI.rc site found at 6627


Design Notes

We added complementary restriction sites on either side of vioC and vioD (BamHI, BglII, HindIII). This allows them to be seperately removed to produce more colours from the operon.


Source

The VioA-E genes were originally from from Chromobacterium voilaceum ATCC 12472 in the pPSX vio+ plasmid. This was kindly provided by John Pemberton; Department of Microbiology and parasitology, University of Queensland, Brisbane, Australia. (Sarovich & Pemberton (2007) Plasmid 57:306-313). The completed gene was designed using GeneDesigner program from DNA2.0 and the codons optimised for E. coli with the help of DNA2.0 (Welch, Villalobos, Gustafsson and Minshull (2009) J. R. Soc. Interface)

Safety Notes

L-Tyrosine

Eye: Causes eye irritation. Skin: Causes skin irritation. May cause dermatitis. May be harmful if absorbed through the skin. Ingestion: May cause irritation of the digestive tract. May be harmful if swallowed. Inhalation: Causes respiratory tract irritation. May be harmful if inhaled. Chronic: May cause liver and kidney damage. Adverse reproductive effects have been reported in animals. Laboratory experiments have resulted in mutagenic effects. Chronic exposure may cause blood effects.

References

Daniel S. Philip, Derek S. Sarovich, John M. Pemberton, Complete sequence and analysis of the stability functions of pPSX, a vector that allows stable cloning and expression of Streptomycete genes in Escherichia coli K12, Plasmid, Volume 62, Issue 1, July 2009, Pages 39-43, ISSN 0147-619X, DOI: 10.1016/j.plasmid.2009.03.002. (http://www.sciencedirect.com/science/article/B6WPF-4VWB194-1/2/b6d8781d5fbc3eb1715f9080eb3594d2) Keywords: Streptomyces; E. coli; Stable vector; Complete sequence; pPSX

RT Journal A1 Welch, Mark A1 Villalobos, Alan A1 Gustafsson, Claes A1 Minshull, Jeremy T1 You're one in a googol: optimizing genes for protein expression YR 2009 FD August 6 JF Journal of The Royal Society Interface JO Journal of The Royal Society Interface SP S467 OP S476 DO 10.1098/rsif.2008.0520.focus VO 6 IS Suppl 4 UL http://rsif.royalsocietypublishing.org/content/6/Suppl_4/S467