Coding

Part:BBa_K5301015

Designed by: Jiaxin Cao   Group: iGEM24_BNU-China   (2024-09-23)
Revision as of 08:21, 30 September 2024 by Caojiaxin (Talk | contribs)


spNW50 is high molecular weight membrane scaffold protein used to produce large nanodiscs.

NW50 is high molecular weight membrane scaffold protein used to produce large nanodiscs.spNW50 could construct nanodiscs with large diameter, and is circularized by SpyTag-Spycatcher.

Characterization

The theoretical molecular weight of NW50 is 123.9kDa, and the actual molecular weight size verified by SDS-PAGE in literature and practical experiments is 150kDa. Due to the large molecular weight of the protein, NW50 is prone to dimerization during actual protein extraction. Protein dimerization can be minimized by adding the detergent Triton X-100 to the refined extraction buffer and reducing the nickel column elution time during the refined extraction process. It should be used as soon as possible after protein extraction to prevent the protein from self-dimerizing after a period of time. A more specific exploration of protein dimerization conditions can be found in the engineering section of 2024BNUChina iGEM. Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 1576
    Illegal SpeI site found at 448
    Illegal PstI site found at 664
    Illegal PstI site found at 1204
    Illegal PstI site found at 1237
    Illegal PstI site found at 1744
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 1576
    Illegal NheI site found at 64
    Illegal SpeI site found at 448
    Illegal PstI site found at 664
    Illegal PstI site found at 1204
    Illegal PstI site found at 1237
    Illegal PstI site found at 1744
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 1576
    Illegal BglII site found at 924
    Illegal BglII site found at 1464
    Illegal BglII site found at 1632
    Illegal BamHI site found at 97
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 1576
    Illegal SpeI site found at 448
    Illegal PstI site found at 664
    Illegal PstI site found at 1204
    Illegal PstI site found at 1237
    Illegal PstI site found at 1744
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 496
    Illegal EcoRI site found at 1576
    Illegal SpeI site found at 448
    Illegal PstI site found at 664
    Illegal PstI site found at 1204
    Illegal PstI site found at 1237
    Illegal PstI site found at 1744
  • 1000
    COMPATIBLE WITH RFC[1000]


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