Coding

Part:BBa_K1152010

Designed by: Ralf Beer, Konrad Herbst, Nikolaos Ignatiadis   Group: iGEM13_Heidelberg   (2013-09-19)

Svp 4'-Phosphopanthetheinyl-transferase


Figure 1: Phenotypes of TOP10 producig indigoidine. As negative control untransformed TOP10 were used.

Svp (Figure 1) encodes for a 4'-Phosphopanthetheinyl-transferase (PPTase) which transfers a 4'-PPT residue from CoA to a conserved serine residue in the T-Domain of NRPS modules, thus activating these enzymes (see Figure 2).

Figure 2: General mechanism of activation the activation of NRP Synthetases from apo- to holo-form.

Figure 3 shows the analytical restiction digest we performed for part validation:

Figure 2: Restriction digest with EcoRI, XbaI, SpeI, PstI.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 173
    Illegal NgoMIV site found at 413
    Illegal NgoMIV site found at 663
    Illegal AgeI site found at 573
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 366

References

  • Takahashi, H., Kumagai, T., Kitani, K., Mori, M., Matoba, Y., & Sugiyama, M. (2007). Cloning and Characterization of a Streptomyces Single Module Type Non-ribosomal Peptide Synthetase Catalyzing a Blue Pigment Synthesis *, 282(12), 9073–9081. doi:10.1074/jbc.M611319200


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