Coding

Part:BBa_K3738028:Design

Designed by: Emily Hagens and Mark Lea   Group: iGEM21_Lethbridge   (2021-10-21)
Revision as of 03:40, 22 October 2021 by Emily Hagens (Talk | contribs) (References)

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MlrA N-terminal 6XHistidine Tag and C-Terminal and Anionic Tag (Basic)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 184
    Illegal BglII site found at 925
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 867


Design Notes

This composite part includes the IPTG-inducible T7 promoter (BBa_J64997), RBS BBa_B0034, Lbu-Cas13a with an N-terminal 6xHistidine Tag and C-Terminal Anionic Tag coding region (BBa_K3738020), and double terminator BBa_B0015.The part is codon-optimized for use in E. coli It is improved from the Lethbridge High School iGEM 2019's Parts BBa_K3001003, BBa_K3001000 and BBa_K3001002 by introducing the anionic MS2 phage-like-particle uptake anionic tag as well as the transcriptional and translational regulators for optimal overexpression and 6XHistidine tag required for nickel affinity chromatography purification.



Source

Source: MlrA from Sphingopyxis sp. USTB-05 part of the mlrABCD gene cassette

References

Saito T, Okano K, Park H-D, Itayama T, Inamori Y, Neilan BA, et al. Detection and sequencing of the microcystin LR-degrading gene, mlrA, from new bacteria isolated from Japanese lakes. FEMS Microbiology Letters. 2003;229(2):271-6. doi: 10.1016/S0378-1097(03)00847-4.

Jiang Y, Shao J, Wu X, Xu Y, Li R. Active and silent members in the mlr gene cluster of a microcystin-degrading bacterium isolated from Lake Taihu, China. FEMS Microbiology Letters. 2011;322(2):108-14. doi: 10.1111/j.1574-6968.2011.02337.x.

Wei J, Huang F, Feng H, Massey IY, Clara T, Long D, et al. Characterization and Mechanism of Linearized-Microcystinase Involved in Bacterial Degradation of Microcystins. Frontiers in Microbiology. 2021;12(527). doi: 10.3389/fmicb.2021.646084.