Composite

Part:BBa_K4849039:Design

Designed by: Devansh Kumar   Group: iGEM23_Edinburgh   (2023-10-12)


MtrCAB + CymA - LvT


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 262
    Illegal BsaI site found at 6111


Design Notes

We designed these parts using the Cynogate assembly (refer IGEM Edinburgh 2023 wiki) Cyanogate CyanoGate is a versatile genetic engineering system developed to address the synthetic biology gap in cyanobacteria. Leveraging the Golden Gate MoClo kit, CyanoGate unites cyanobacteria with plant and algal systems, offering a standardized modular cloning approach. Developed by the McCormick lab at the University of Edinburgh, CyanoGate facilitates the generation of knock-outs, integrations, and multigene expression systems in cyanobacteria. Cyanobacteria, crucial for their ecological importance and potential biotechnological applications, have been underutilized in synthetic biology. CyanoGate aims to bridge this gap by providing a suite of genetic parts and acceptor vectors, enabling integrative or replicative transformation. Tested in model cyanobacteria, Synechocystis sp. PCC 6803 and Synechococcus elongatus UTEX 2973, CyanoGate demonstrates its applicability for efficient genetic modification. The importance of cyanobacteria in biotechnology, from pharmaceuticals to biophotovoltaic devices, underscores the need for robust synthetic biology tools. Despite progress, cyanobacteria lag behind in synthetic biology compared to other systems. CyanoGate addresses this challenge by offering an easy-to-use system that integrates with existing standards, enhancing scalability and accelerating the "design, build, test, and learn" cycle. The syntax of level 0 parts was adapted for prokaryotic cyanobacteria, addressing typical cloning needs in cyanobacterial research. New level 0 parts were assembled from various sources. Level 1, M, and P acceptor vectors from the MoClo Plant Tool Kit facilitated the assembly of level 0 parts in a level 1 vector, with up to seven level 1 modules in level M. Level M assemblies could be further combined into level P and cycled back into level M for larger multimodule vectors if needed. Vectors exceeding 50 kb in size, assembled by MoClo, have been reportedModules from level 1 or level P could be assembled in new level T vectors designed for cyanobacterial transformation. Both UTEX 2973 and Synechocystis produced recombinants through electroporation or conjugation methods with level T vectors, with a preference for the conjugation approach in the outlined work.



Source

Shewanella oneidensis MR-1 (NCBI:txid211586)

References