Part:BBa_K3738028:Design
MlrA N-terminal 6XHistidine Tag and C-Terminal and Anionic Tag (Basic)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 184
Illegal BglII site found at 925 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.