Part:BBa_K2578720

Protein coding sequence for Mouse metallothionein 2 (Mt2)

The part BBa_K2578720 is the composite part of the Mus musculus metallothionein 2 synthesising gene(Mt2). The genomic sequence of the Mt2 gene is referenced from NCBI (Gene ID 17750)[1].

The Mt2 gene, registered as BBa_K2578721, is designed to be the composite part BBa_K2578720 by adding consecutive promoter J23100, strong RBS B0032 and double terminator B0015.

Metallothioneins (MTs) are small cysteine-rich proteins made of 61-68 amino acids which can be found in a broad range of organisms, including both eukaryotes and prokaryotes. The MTs are expressed as intracellular protein. [5] MTs are mainly responsible for metalloregulation in cells of living organisms. The rich Cys domains in MTs allow the non-covalent binding of trace metals such as cadmium, lead, copper and mercury, etc. [6,7]

The Mt2 gene has similar ability to Mt1 gene as they are equivalent proteins. They have strong binding affinity to heavy metals[8] and serve severe function like metal homeostasis and metabolism and protect cell from oxidative stress of zinc ion in mouse liver[8,9]. But when compared to the metal binding ability to metal ions, results shown that Mt2 has better binding ability of Zinc 2+ ions while Mt1 gene has better binding ability of copper 1+ ions and cadmium 2+ ions.[10]

We failed to submit the part to the iGEM community as there are multiple restriction enzyme site and introns in the gene of interest. Therefore, there is an incompletion in the subproject on the Mt1 gene.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 425
  Illegal SpeI site found at 453
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 425
  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal SpeI site found at 453
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 425
  Illegal BamHI site found at 576
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 425
  Illegal SpeI site found at 453
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 425
  Illegal SpeI site found at 453
  
• 1000
  COMPATIBLE WITH RFC[1000]