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Part:BBa_K1149002

Designed by: Margarita Kopniczky   Group: iGEM13_Imperial_College   (2013-09-13)
Revision as of 17:27, 15 September 2013 by Margarita K (Talk | contribs)

pelB-EstCS2-his expression (GFP reporter)

This part expresses a plastic degradation enzyme and sfGFP under the control of arabinose inducible promoter.

The enzyme is the estCS2 from BBa_K892012 previous biobrick by Washington 2012 team, that encodes carboxylesterase that breaks down ester bonds in polyurethane. We have added a pelB seretion tag to the enzyme so that it can be secreted extacellularly. We also added a 6xHistidine tag for Western blotting and protein purification purposes.

Theoretical pathway of PUR degradation by esterase: PUR_esterase_theoretical_pathway.jpg


references:

[http://www.selu.edu/acad_research/depts/biol/faculty/publications/pdf/2012/howard2012.pdf Howard, Gary 2012 pg:371 -3, Polyurethane Biodegradation, Environmental Science]

[http://www.microbialcellfactories.com/content/10/1/41 A novel family VII esterase with industrial potential from compost metagenomic library. Microbial Cell Factories 2011]

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
    Illegal BamHI site found at 692
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 210
    Illegal NgoMIV site found at 958
    Illegal NgoMIV site found at 1192
    Illegal AgeI site found at 2131
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1006


[edit]
Categories
//cds/biosynthesis
Parameters
None