Translational_Unit

Part:BBa_K2848004

Designed by: Lars Hoeing, Liv Paul, Luca Schelle   Group: iGEM18_Stuttgart   (2018-10-09)


Pre-NisA under control of Arabinose promotor

Pre-NisA (BBa_K2848003) under control of the araBAD promoter. The following image shows the theoretical construct of the composite BioBrick.

Figure 1: Theoretical construct of the composite BioBrick, containing pre-Nisin A, with YAAY- and His6-tag and an inducible arabinose promoter (araBAD). Visualized by SnapGene.

Sequence of the BioBrick was confirmed by sanger sequencing.

DH5 alpha E. coli were transformed with this Brick to produce the modified nisin. Cell lysate was purified with a His-Trap column (ÄKTA). The elution was analyzed by SDS-PAGE. The obtained gel was stained using silver staining.

Figure 2: Gel from SDS-PAGE, stained using silver staining.

Thus, the modified nisin was produced successfully.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


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