Part:BBa_K4035003

Dimerization of the copper metallothionein 1 : CUP1-(GGGGS)4-CUP1

Copper metallotionein CUP1 (BBa_M45090) is a protein responsible for copper binding protein in the yeast Saccharomyces cerevisiae. Normally expressed intracellularly, CUP1 was fused to the A-agglutinin-binding subunit Aga2 (BBa_K416000) in order to be displayed on the outter yeast cell surface. In order to improve the copper fixation efficiency in yeast, CUP1 was linked to a second sequence of the same protein. The linker is a flexible linker and is made of four successive GGGGS amino acid sequence. This fusion protein also contained a V5 tag in order to check its expression by Western Blot and Immunostaining. The expression was under the control of the Gal1 promoter, so that the protein was expressed only in the presence of galactose.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 90
  Illegal PstI site found at 408
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 90
  Illegal PstI site found at 408
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 235
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 90
  Illegal PstI site found at 408
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 90
  Illegal PstI site found at 408
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 84