Composite

Part:BBa_K2123204:Design

Designed by: Maria Clara Tavares Astolfi   Group: iGEM16_UFAM-UEA_Brazil   (2016-10-13)
Revision as of 10:17, 22 October 2016 by Mctastolfi (Talk | contribs)

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Bioaccumulator device: Strong promoter + OmpA fused to Synthetic Phytochelatin + B0015


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1493
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1091
    Illegal NgoMIV site found at 1665
    Illegal NgoMIV site found at 2902
    Illegal NgoMIV site found at 2964
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1084


Design Notes: In silico study

To construct our final bioaccumulation device, we followed the steps below:

1st Step:

800px-UFAM_UEA_FTQ_DESIGN_2.png"

Phytochelatin digestion with EcoRI + SpeI releasing Lpp-OmpA with NdeI and XbaI. To linearize Lpp-OmpA (2526 bp) and release the promoter (~83 bp).

3rd Step:

800px-UFAM_UEA_FTQ_DESIGN_3.png</center.

Digestion of first step construction, resulting the PO construction (JK26 promoter + Lpp-OmpA) digested with EcoRI + SpeI, releasing a 564 bp insert. The PT (Phytochelatin + Terminator) construction was linearized with EcoRI + XbaI, releasing insert of approximately 2317 bp.

Finally we had our bioaccumulator construction!!