Tag

Part:BBa_K1486027

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

R.reniformis luciferase + ADH1 terminator + Kanamycin resistance

This part is a tag Renilla renifomis luciferase (rLuc) associated with a 13 amino acids linker, along a terminator and kanamycin resistance. We used it as a control of the expression and activity of luciferase in the budding yeast S.cerevisiae, by targeting the gene Pbs2. 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 PstI site found at 1490
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1490
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1490
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1490
  • 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
resistanceKanamycin & chloramphenicol (backbone)