Composite

Part:BBa_K4192132

Designed by: Jiahui Li   Group: iGEM22_CAU_China   (2022-09-30)
Revision as of 17:26, 9 October 2022 by Subconsciousness (Talk | contribs)


Plac-RBS-yf1(N37C)-RBS-fixJ-PfixK2-RBS-mcherry

The gene yf1, fixJ and PfixK2 make up the system of blue light regulation. Under dark conditions, phosphate groups are transferred from Yf1 protein to FixJ protein, and the phosphorylated FixJ protein activates the PfixK2 promoter, which in turn activates downstream gene expression. Under light induction, the phosphorylation of Fixj protein was blocked and the expression of genes regulated by the PfixK2 promoter was suppressed.The mCherry protein emits red fluorescence to characterize the efficiency of control elements in regulating gene expression. We mutated position 37 N of YF1 to C to improve its response to blue light and better regulate gene expression.


Usage

Light control system can regulate gene expression by light intensity. MCherry has an excitation wavelength of 580nm and an emission wavelength of 610nm. Quantitative detection of mCherry can be achieved by detecting the optical signal at 610nm. This composition part examines the ability of the light control system containing the yf1 mutation to control gene expression and compares it with the normal light control system (BBa_ K4192130).

Biology

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 210
    Illegal SpeI site found at 1361
    Illegal SpeI site found at 2257
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 210
    Illegal SpeI site found at 1361
    Illegal SpeI site found at 2257
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 210
    Illegal SpeI site found at 1361
    Illegal SpeI site found at 2257
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 210
    Illegal SpeI site found at 1361
    Illegal SpeI site found at 2257
    Illegal NgoMIV site found at 764
    Illegal NgoMIV site found at 836
    Illegal NgoMIV site found at 926
    Illegal NgoMIV site found at 944
    Illegal NgoMIV site found at 1450
    Illegal NgoMIV site found at 1743
    Illegal NgoMIV site found at 1837
    Illegal AgeI site found at 478
    Illegal AgeI site found at 1618
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1507
    Illegal BsaI.rc site found at 377


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Categories
Parameters
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