Coding

Part:BBa_K929202:Design

Designed by: Potsdam Bioware 2012 iGEM   Group: iGEM12_Potsdam_Bioware   (2012-09-24)

EGF-Receptor_3rd domain with PelB leadersequence, His-tag and C-terminal Sortase motif


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 894
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 664
    Illegal AgeI site found at 1342
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

UP12 BBa K929202 design3.png

Source

The Biobrickpart was designed with restriction sites according to the RFC25 suffix and prefix. We cloned the desired part into the Biobrick containing CMV promoter (BBa_I712004). The resulting Biobrick (BBa_K929202) corresponds to the original RFC10 prefix and suffix. But the desired fusionparts PelB, EGF-Receptor_3rd domain, His-tag and C-terminal sortase motif have 2 additional enclosing restriction sites according to the RFC25. That means, that the Part 'PelB, EGF-Receptor_3rd domain, His-tag and C-terminal sortase motif' could be cut out with the enzyms NgoMIV and AgeI

References

  • Leung K. Cy5.5-Ac-Cys-ZEGFR:1907. 2012 Jun 21 [updated 2012 Sep 07]. Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda(MD): National Center for Biotechnology Information (US); 2004-2012. [http://www.ncbi.nlm.nih.gov/pubmed/22973578 PubMed PMID: 22973578]
  • Zhixiang Wang (2012). Mutual Regulation of Receptor-Mediated Cell Signalling and Endocytosis: EGF Receptor System as an Example, Molecular Regulation of Endocytosis, Brian Ceresa (Ed.), ISBN: 978-953-51-0662-3, InTech, [http://www.intechopen.com/books/molecular-regulation-of-endocytosis/mutual-regulation-of-receptor-mediated-cell-signalling-and-endocytosis-egf-receptor-system-as-an-exa InTech]
  • Abdullah SE, Haigentz M Jr, Piperdi B. Dermatologic Toxicities from Monoclonal Antibodies and Tyrosine Kinase Inhibitors against EGFR: Pathophysiology and Management. Chemother Res Pract. 2012;2012:351210. Epub 2012 Sep 11. PubMed PMID: 22997576; [http://www.ncbi.nlm.nih.gov/pubmed/22997576 PMCID: PMC3446637]
  • Ayako Kamei, Paul S. Hauser, Jennifer A. Beckstead, Paul M.M. Weers, Robert O. Ryan Expressed protein ligation using an N-terminal cysteine containing fragment generated in vivo from a pelB fusion protein. Published in final edited form as: Protein Expr Purif. 2012 June; 83(2): 113–116. Published online 2012 April 1. doi: 10.1016/j.pep.2012.03.014 [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1963442/ PMCID: PMC1963442]