Part:BBa_K4035001

CUP1 fused to Aga2 and tagged with a V5 epitope

This protein is made of the copper metallotionein CUP1 (BBa_M45090) fused to the A-agglutinin-binding subunit Aga2 (BBa_K416000) at its N-terminal and to the V5 tag at its C-terminal.

Usage and Biology

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) that attaches to the yeast cell wall through disulfide bonds to Aga1p. This leading to the presence of CUP1 on the outter membrane of the cell. 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 319
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 376
  Illegal PstI site found at 319
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 562
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 319
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 319
  
• 1000
  COMPATIBLE WITH RFC[1000]