Plasmid

Part:BBa_K3600013

Designed by: yasmine koubaa   Group: iGEM20_EPFL   (2020-10-27)
Revision as of 02:40, 28 October 2020 by Sspoerri (Talk | contribs) (→‎Usage and Biology)


Lys2-HSP12pr-mScarleti cassette

This cassette is the product of a two part (BBa_K3600006, BBa_K3600012) Golden Gate BsmBI assembly.

Usage and Biology

This cassette will be integrated into the yeast genome to form a reporter strain that produces a fluorescent signal when the yeast cell is under environmental stress.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal XbaI site found at 1193
    Illegal XbaI site found at 1618
    Illegal XbaI site found at 2091
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
    Illegal NotI site found at 1172
    Illegal NotI site found at 3078
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal suffix found in sequence at 2661
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal BglII site found at 1636
    Illegal BglII site found at 2532
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal XbaI site found at 1193
    Illegal XbaI site found at 1618
    Illegal XbaI site found at 2091
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1184
    Illegal EcoRI site found at 1914
    Illegal XbaI site found at 1193
    Illegal XbaI site found at 1618
    Illegal XbaI site found at 2091
    Illegal SpeI site found at 3059
    Illegal PstI site found at 3068
    Illegal NgoMIV site found at 3016
  • 1000
    COMPATIBLE WITH RFC[1000]


Experiments

1mg/L Atrazine in methanol was added to overnight cultures to test if the expression of mScarletI could be induced. The final concentration of methanol is 1%. The signal was quantified using imageJ. The fluorescence signals were corrected for the background. The mean signal (n=15) for wildtype cells in the same setup was found to be at 12.1928 after correcting for the background.


[edit]
Categories
Parameters
None