Composite

Part:BBa_K5301024:Design

Designed by: Xiaowen Sun   Group: iGEM24_BNU-China   (2024-09-24)


Multi-polymer MSP, which refers to large circular MSPs through end-to-end connections of multiple MS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 3141
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 3141
    Illegal NotI site found at 1720
    Illegal NotI site found at 4038
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1239
    Illegal BglII site found at 2323
    Illegal BglII site found at 2695
    Illegal BglII site found at 3557
    Illegal BglII site found at 3929
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 3141
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 3141
    Illegal AgeI site found at 1035
    Illegal AgeI site found at 1828
    Illegal AgeI site found at 2220
    Illegal AgeI site found at 2980
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We carefully designed the joints of three MSP segments to ensure that a single MSP does not undergo self-cyclization or dimerization, but can form the correct multi-polymerized MSP. Finally, mCherry emits red fluorescence after covalent interaction occurs at the last pair of linkers-SpyCatcher/Tag, indicating the successful construction of MSP.


Source

SpyCatcher/Tag are derived from Streptococcus pyogenes fibronectin-binding protein, FbaB. SdyCatcher/Tag are derived from Streptococcus dysgalactiae protein, CnaB. SnoopCatcher/Tag are derived from RrgA, an adhesin from Streptococcus pneumoniae. mCherry [1-10] and [11] come from mCherry fluorescent protein. NW15 is the truncated form of NW30, which is originated from a set of α-helixes derived from apolipoprotein A-1 (ApoA1).

References