Coding

Part:BBa_K4960039

Designed by: Ziying Wang   Group: iGEM23_NUDT-CHINA   (2023-10-11)
Revision as of 13:31, 11 October 2023 by WZY24 (Talk | contribs)


pPVC promoter->pvc1-pvc12->pvc13_NTD-3*EAAAK-CKGGRAKDC-3*EAAAK-pVc13_CT->pvc14-pvc16

A flexible linker which can influence the structure of tail fiber connected to the CKGGRAKDC.

Special Design

Use the CKGGRAKDC to target fat cells. The different linker(1*EAAAK or 3*EAAAK) on both sides of the CKGGRAKDC can affect the structure and function of the tail protein. By altering the linker on both sides of the CKGGRAKDC, we explored the most suitable structure of the tail protein for recognition and bond. At the same time, because of the existence of linker, we can use Golden Gate to replace the sequences in the middle of linker more efficiently than enzyme cutting and enzyme linking.

Sequence and Feature

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7406
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2407
    Illegal BglII site found at 11264
    Illegal BglII site found at 14099
    Illegal BglII site found at 17020
    Illegal XhoI site found at 2691
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 11792
    Illegal NgoMIV site found at 17551
    Illegal NgoMIV site found at 17690
    Illegal AgeI site found at 6704
    Illegal AgeI site found at 9641
    Illegal AgeI site found at 10327
    Illegal AgeI site found at 10468
    Illegal AgeI site found at 11576
    Illegal AgeI site found at 17989
    Illegal AgeI site found at 19677
  • 1000
    COMPATIBLE WITH RFC[1000]

Tube:pvc1, pvc5, pvc7
Sheath:pvc2, pvc3, pvc4
Sheath terminator:pvc15
Tail fbres:pvc13_NTD-3*EAAAK-CKGGRAKDC-3*EAAAK -pVc13_CTD
Baseplate:pvc9, pvc11, pvc12
Spike:pvc8, pvc10
Assembly method:3A Assembly

References

[1]Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F. Programmable protein delivery with a bacterial contractile injection system. Nature. 2023 Apr;616(7956):357-364. doi: 10.1038/s41586-023-05870-7. Epub 2023 Mar 29. PMID: 36991127; PMCID: PMC10097599.
[2]Kolonin MG, Saha PK, Chan L, Pasqualini R, Arap W. Reversal of obesity by targeted ablation of adipose tissue. Nat Med. 2004 Jun;10(6):625-32. doi: 10.1038/nm1048. Epub 2004 May 9. PMID: 15133506.

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