Coding

Part:BBa_K4259000

Designed by: Jennifer Adami   Group: iGEM22_Leiden   (2022-09-26)


Cup1 for E. coli

Cup1 is a gene in Saccharomyces cerevisiae that encodes for metallothionein (MT), a metal-binding protein.


Usage and Biology

Metallothionein plays an important role in intracellular metal distribution and the protection of the cell against heavy metal toxicity and oxidative stress. MT has a high metal binding affinity. Due to its role in metal reduction, previous studies have investigated its potential for the production of metal nanoparticles (NPs). This was done by integrating MT in various biological platforms.1 Thus, we expressed cup1 in an engineered Escherichia Coli (E. coli), with the aim of producing gold and silver NPs.

Design

The sequence was codon optimized for E.coli. Two overhang sequences with the restriction enzymes BamHI and NdeI were designed and added to the sequence. This part is therefore compatible with vectors that have those two restriction sites. In the Binanox project, the vector pET-16b was used.


References

1. Yuan Q, Bomma M, Xiao Z. Enhanced Silver Nanoparticle Synthesis by Escherichia Coli Transformed with Candida Albicans Metallothionein Gene. Materials (Basel). 2019 Dec 12;12(24):4180. doi: 10.3390/ma12244180. PMID: 31842386; PMCID: PMC6947575.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 123
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None