Plasmid_Backbone

Part:BBa_K1391017

Designed by: Jeremy Gam   Group: iGEM14_MIT   (2014-10-17)

MAV1212-RFP

RFP inside a MAV1212 backbone.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found at 1252
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2721
    Illegal XbaI site found at 2694
    Illegal SpeI site found at 2198
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 2682
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 1252
    Illegal EcoRI site found at 2721
    Illegal SpeI site found at 2198
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 2682
    Illegal NotI site found at 402
    Illegal NotI site found at 1258
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 1252
    Illegal EcoRI site found at 2721
    Illegal BglII site found at 3182
    Illegal BamHI site found at 2205
    Illegal BamHI site found at 2700
    Illegal XhoI site found at 394
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 1252
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2721
    Illegal XbaI site found at 2694
    Illegal SpeI site found at 2198
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 2682
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 1252
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 2721
    Illegal XbaI site found at 1267
    Illegal XbaI site found at 2694
    Illegal SpeI site found at 2198
    Illegal PstI site found at 730
    Illegal PstI site found at 1011
    Illegal PstI site found at 2682
    Illegal NgoMIV site found at 645
    Illegal NgoMIV site found at 926
    Illegal AgeI site found at 774
    Illegal AgeI site found at 1908
    Illegal AgeI site found at 2020
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 2465
    Illegal BsaI.rc site found at 3083



MAV1212 was the backbone we used to construct our L7ae repression system. Before coming to any conclusions about the success of our constructs, we needed to make sure that the L7ae/k-turn system worked correctly. To do this we expressed k-turn:eGFP with and without the presence of constitutive L7ae. We used eBFP as our normalizing transfection marker.


L7ae Repression of 2x-Kturn-GFP. In the absence of L7ae, eBFP2 and GFP fluorescence show a positive relationship, as expected when co-expressing two constitutive fluorophores. However, upon the addition of L7ae into the system, GFP production is almost completely silenced. This indicates that our L7ae / 2x-Kturn system is viable to implement in our high sensors.

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Categories
Parameters
None