Composite

Part:BBa_K2350021:Design

Designed by: YA-XUAN YANG   Group: iGEM17_NYMU-Taipei   (2017-10-22)


R0010-B0034-Holin-B0010-B0012-J23106-B0034-Endolysin-B0010-B0012-J23118-B0034-NrtA-B0015


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1035
    Illegal NheI site found at 1058
    Illegal NheI site found at 1757
    Illegal NheI site found at 1780
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2990
    Illegal BamHI site found at 2360
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1362
    Illegal AgeI site found at 1432
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

　　In our 2017 NYMU project, the endolysin and holin both are from enterobacteria phage T4, for the reason that we don’t know whether endolysin and holin from different species could cooperate well.

Source

　　The endolysin and holing we used are from iGEM released parts, BBa_K112806 and BBa_K112000, respectively. NrtA is from cyanobacteria Synechocystis sp. PCC 6803.

References

  • Koropatkin NM1, Pakrasi HB, Smith TJ. Atomic structure of a nitrate-binding protein crucial for photosynthetic productivity. 2006 Jun 27;103(26):9820-5. Epub 2006 Jun 15.
  • Ye Wang, Wenbin Li, Yaeesh Siddiqi, James R. Kinghorn, Shiela E. Unkles and Anthony D. M. Glass. Evidence for post-translational regulation of NrtA, the Aspergillus nidulans high-affinity nitrate transporter. DOI: 10.1111/j.1469-8137.2007.02135.x • Source: PubMed
  • Alexova R, Haynes PA, Ferrari BC, Neilan BA. Comparative protein expression in different strains of the bloom-forming cyanobacterium Microcystis aeruginosa. 2011 Sep;10(9):M110.003749. doi: 10.1074/mcp.M110.003749. Epub 2011 May 24.
  • Omata T, Ohmori M, Arai N, Ogawa T. Genetically engineered mutant of the cyanobacterium Synechococcus PCC 7942 defective in nitrate transport. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6612-6.
  • Naureen Akhtar, Eugenia Karabika, James R. Kinghorn, Anthony D.M. Glass, Shiela E. Unkles, and Duncan A. Rouch. High-affinity nitrate/nitrite transporters NrtA and NrtB of Aspergillus nidulans exhibit high specificity and different inhibitor sensitivity. Microbiology. 2015 Jul; 161(Pt 7): 1435–1446. doi: 10.1099/mic.0.000088
  • James R. Kinghorn, Joan Sloan, Ghassan J. M. Kana'n, Edisio R. DaSilva, Duncan A. Rouch, and Shiela E. Unkles. Missense Mutations That Inactivate the Aspergillus nidulans nrtA Gene Encoding a High-Affinity Nitrate Transporter. Genetics. 2005 Mar; 169(3): 1369–1377. doi: 10.1534/genetics.104.036590
  • Maeda S, Omata T. Substrate-binding lipoprotein of the cyanobacterium Synechococcus sp. strain PCC 7942 involved in the transport of nitrate and nitrite. J Biol Chem. 1997 Jan 31;272(5):3036-41.
  • 2008 UC Berkeley Team: http://2008.igem.org/Team:UC_Berkeley
  • 2014 Peking Team: http://2014.igem.org/Team:Peking/Suicide