Composite

Part:BBa_K4035000

Designed by: Anissa Hammi   Group: iGEM21_EPFL   (2021-08-30)
Revision as of 15:15, 30 August 2021 by Registry (Talk | contribs)

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Expression of CUP1 on the extracellular membrane of S. cerevisiae

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. 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
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//binding/metal
//cds/membrane/extracellular
//chassis/eukaryote/yeast
Parameters
biologyCUP1
chassisS.cerevisiae
controlpCTcon2V5, WT EBY100
directionForward
functioncopper binding protein