Composite

Part:BBa_K1493602

Designed by: Teresa Robert Finestra   Group: iGEM14_Wageningen_UR   (2014-10-06)

pLacI + Kis Antitoxin (Kis-Kid Toxin-Antitoxin System)

Kis (antitoxin) and Kid (toxin) are part of the parD stability system of plasmid R1 acting as a pre-segregation mechanism to ensure plasmid proliferation. Kid breaks down single stranded messenger RNAs. By degradation of something as vital as messenger RNA, this toxin does not differ in function with gram positive and gram negative bacteria. In nature, Kid causes cells to divide slower to ensure that every daughter cell receives the R1 plasmid. The Kis-Kid interaction has proven to be a 1:2 ratio [1, 2].

The Kis antitoxin was amplified and elongated form the pAB24 plasmid, as well as, the Kid toxin. iGEM Wageningen team 2014 achieved successful cloning of Kid (Toxin) in a low copy number plasmid (psB3K3) in combination with the antitoxin in E. coli JM109 strain. If you are interested in the glycerol stock visit the iGEM Wageningen Team 2014 wiki and/or contact us.

This part is regulated by the LacI promoter.


References:

1. de la Cueva-Méndez, G. and B. Pimentel, Gene and cell survival: lessons from prokaryotic plasmid R1. EMBO reports, 2007. 8(5): p. 458-464.

2. Ruiz-Echevarría, M.J., et al., Kid, a small protein of the parD stability system of plasmid R1, is an inhibitor of DNA replication acting at the initiation of DNA synthesis. Journal of molecular biology, 1995. 247(4): p. 568-577.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 344
  • 1000
    COMPATIBLE WITH RFC[1000]


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