Coding

Part:BBa_K2201207

Designed by: Yannic Kerkhoff   Group: iGEM17_Bielefeld-CeBiTec   (2017-10-20)
Revision as of 07:44, 25 October 2017 by Yanker (Talk | contribs)

Expression plasmid of an aminoacyl-tRNA synthetase for AzoPhe

aminoacyl-tRNA synthetase for incorporating AzoF at the amber codon

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Figure 1: Plasmid map of the BioBrick K2201207 coding for a AzoF-tRNA-synthetase system based on the ONBY-Part (K1416000). The ONBY-RS was removed by PCR and then a gene containing the AzoF-RS and matching overlaps sides was integrated through Gibson assembly.

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Figure 2: History of the plasmid construction of K2201207.

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Figure 3: Alignment of the protein sequence of the AzoF-RS and the M. jannaschii TyrRS.

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Figure 4: Structure of AzoF in cis- and trans-conformation. Switching is possible via irradiation.

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Figure 5: Absorption spectrum of AzoF in LB media after irradiation with light of 367 nm and 465 nm wavelength. The black line shows the typical absorption of AzoF in a mixed state, the trans-conformation is blue and green and the other lines show the absorption spectrum in the cis-conformation.

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Figure 6: Absorption at 476 nm (indicator for lycopene) normalized to the OD600 (indication for the cell density) to calculate the relative lycopene production of each crtI variant cultivated with AzoF in cis- and trans-conformation.

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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 1164
    Illegal BamHI site found at 1170
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1196
    Illegal NgoMIV site found at 1656
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 295
    Illegal SapI.rc site found at 327
    Illegal SapI.rc site found at 1120


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