Plasmid

Part:BBa_K1391027

Designed by: Jing Wei "Raymond" Liu   Group: iGEM14_MIT   (2014-10-17)

Low sensor: eBFP2_miR-144_miR-181c

Our completed low sensor for detection of miR-146a AND miR-181c, which contains target sites for both miRNA sequences and thus requires low concentrations of both miRNAs for successful activation. Uses eBFP2 as a fluorescent reporter.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal NotI site found at 402
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal BglII site found at 1694
    Illegal BglII site found at 3481
    Illegal XhoI site found at 394
    Illegal XhoI site found at 2093
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1085
    Illegal EcoRI site found at 2314
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 1440
    Illegal PstI site found at 1945
    Illegal NgoMIV site found at 645
    Illegal NgoMIV site found at 926
    Illegal NgoMIV site found at 1828
    Illegal AgeI site found at 774
    Illegal AgeI site found at 1206
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3131
    Illegal BsaI.rc site found at 3382


A


Two low sensors. We co-transfected the same low sensors from the previous Single Input miRNA Sensor Experiments using red as our transfection marker and blue as our output. Given the results of our previous experiments we hoped that the sensors, when exposed to the inhibiting siRNA, would show a more repressed eBFP2 output. However, the cytometry results in figure (A) show that there was still no difference between the 'on' and 'off' state.

[edit]
Categories
Parameters
None