Tag

Part:BBa_K1486033

Designed by: EPFL iGem team 2014   Group: iGEM14_EPF_Lausanne   (2014-09-27)

R.reniformis luciferase + ADH1 terminator + CaUra3

This part is a tag Renilla renifomis luciferase (rLuc) associated with a 13 amino acids linker, along a terminator and a Ura3 cassette (allows selection on media lacking uracyl). We used it as a control of the expression and activity of luciferase in the budding yeast S.cerevisiae, by targeting the gene Hog1. The luciferase is the one from the organism Renilla reniformis, needs the substrate coelenterazine and is ATP independent. Coelenterazine has a spontaneous hydrolysis reaction, and so emits light even without luciferase.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1640
    Illegal PstI site found at 1329
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1640
    Illegal PstI site found at 1329
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1640
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1640
    Illegal PstI site found at 1329
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1640
    Illegal PstI site found at 1329
  • 1000
    COMPATIBLE WITH RFC[1000]


Reference

Studies of Dynamic Protein-Protein Interactions in Bacteria Using Renilla Luciferase Complementation Are Undermined by Nonspecific Enzyme Inhibition: Stavroula K. Hatzios, Simon Ringgaard, Brigid M. Davis, Matthew K. Waldor.

Source

The coding sequence of this luciferase was generously given by Dr. Matthew Waldor's laboratory.

[edit]
Categories
//cds/reporter
//cds/selectionmarker/antibioticresistance
//chassis/eukaryote/yeast
//direction/forward
//function/reporter/light
Parameters
biology
chassisSaccharomyces Cerevisiae
colorGreen
directionForward
emission480
functionTag
resistanceUracil deprived medium