Composite
Q-Sensor

Part:BBa_K4755013

Designed by: joseph drake   Group: iGEM23_UAlberta   (2023-10-11)


Cameleon-Q-Sensor (Synthesized via IDT)

This was our team's first attempt at creating the Cameleon-Q-Sensor. It was synthesized as a single fragment by Integrated DNA Technologies (IDT) and delivered in pUCIDT_Amp+ backbone. The upstream regulatory region was meant to be identical to that of the pET22B(+) backbone, unfortunately, this does not appear to be the case, as the RBS region has been altered slightly. Luckily, the T7 promoter and Lac Operator regions are fully intact and match those present in the pET22B(+) backbone.

There are seven domains present in this composite part: 1. T7 promoter, 2. Lac operator, 3. pET22B(+) RBS (altered slightly), 4. mScarlet (BBa_K4755005), 5. Cameleon (BBa_K4755007), 6. ShadowR (BBa_K4755006), and 7. 6x Histidine Tag. We included four restriction enzyme sites that were meant to allow rapid replacement of mScarlet, Cameleon, and ShadowR with similar protein coding sequences. Those are: NdeI - mScarlet - XbaI - Cameleon - NsiI - ShadowR - HindIII - His Tag.

The ultimate aim of this part was to enable fast and accurate assessments of FRET-quenching between a variety of fluorescent proteins and chromoproteins. Once a suitable pair had been identified, it would be trivial to replace Cameleon with any other domain required to induce association of the fluorescent protein and chromoprotein in your particular experiment.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 131
    Illegal EcoRI site found at 1010
    Illegal EcoRI site found at 1445
    Illegal EcoRI site found at 1804
    Illegal XbaI site found at 797
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 131
    Illegal EcoRI site found at 1010
    Illegal EcoRI site found at 1445
    Illegal EcoRI site found at 1804
    Illegal NheI site found at 536
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 131
    Illegal EcoRI site found at 1010
    Illegal EcoRI site found at 1445
    Illegal EcoRI site found at 1804
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 131
    Illegal EcoRI site found at 1010
    Illegal EcoRI site found at 1445
    Illegal EcoRI site found at 1804
    Illegal XbaI site found at 797
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 131
    Illegal EcoRI site found at 1010
    Illegal EcoRI site found at 1445
    Illegal EcoRI site found at 1804
    Illegal XbaI site found at 797
    Illegal NgoMIV site found at 207
    Illegal AgeI site found at 1305
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 46
    Illegal SapI site found at 862


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Categories
Parameters
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