Composite

Part:BBa_K3278000

Designed by: Ailin Zhou   Group: iGEM19_ShanghaiTech_China   (2019-10-13)


pDawn-mEGFP

This part can characterize the light control efficiency of pDawn system. PDawn is a blue light control system which can start the expression of the target gene. With the target gene usually being proteins or RNAs, it may be relatively difficult to characterize its expression level. Thus, we construct the pDawn+mEGFP by using homologous recombination to show the light control efficiency by fluorescence intensity. Registers can use this part in their own plasmid and bacteria to test its actual efficiency in varied situation.

pDawn with T7 promoter is abbreviated as T7c below. pDawn with tac promoter is abbreviated as tac below. pDawn with modified tac promoter is abbreviated as mtac below. pDawn with YF2 is abbreviated as YF2 below.

[Go for the protocol of characterization:https://2019.igem.org/Team:ShanghaiTech_China/LightControl]

pDawn-mEGFP-0 hour
Fig1. pDawn-mEGFP-0 hour


pDawn-mEGFP-6 hour
Fig2. pDawn-mEGFP-6 hour


pDawn-mEGFP-12 hour
Fig3. pDawn-mEGFP-12 hour


Dark to light-12 hour
Fig4. Dark to light-12 hour


pDawn-mEGFP-24 hour
Fig5. pDawn-mEGFP-24 hour


Dark to light-24 hour
Fig6. Dark to light-24 hour


pDawn-mEGFP-30 hour
Fig7. pDawn-mEGFP-30 hour


Dark to light-30 hour
Fig8. Dark to light-30 hour


pDawn-mEGFP-36 hour
Fig9. pDawn-mEGFP-36 hour


Dark to light-36 hour
Fig10. Dark to light-36 hour




To see more changes on pDawn, go to the websites below.
pDawn-mEGFP: https://parts.igem.org/Part:BBa_K3278000
pDawn with T7 promoter: https://parts.igem.org/Part:BBa_K3278001
pDawn with mtac promoter: https://parts.igem.org/Part:BBa_K3278002
pdawn with tac promoter: https://parts.igem.org/Part:BBa_K3278005
pDawn with YF2: https://parts.igem.org/Part:BBa_K3278006


References:
[1] From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression. Robert Ohlendorf1, Roee R. Vidavski, Avigdor Eldar, Keith Moffat, and Andreas Möglich. Journal of Molecular Biology, 2012, Vol.416, pp. 534-542.
[2] Design and Signaling Mechanism of Light-Regulated Histidine Kinases. Andreas Möglich, Rebecca A. Ayers and Keith Moffat. Biophysical Journal, 2009, Vol.96 (3), pp. 1433-1444.
[3] Oxygen-Regulated In Vitro Transcription of Rhizobium meliloti nifA and fixK Genes. JEAN-MARC REYRAT,' MICHEL DAVID,' CASIMIR BLONSKI,2 PIERRE BOISTARD,' AND JACQUES BATUT'*. Journal of Bacteriology, 1993, pp. 6867-6872.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2148
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 46
    Illegal NgoMIV site found at 178
    Illegal NgoMIV site found at 272
    Illegal NgoMIV site found at 565
    Illegal NgoMIV site found at 1059
    Illegal NgoMIV site found at 1077
    Illegal NgoMIV site found at 1167
    Illegal AgeI site found at 397
    Illegal AgeI site found at 1525
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1620
    Illegal BsaI.rc site found at 508


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Categories
Parameters
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