Composite

Part:BBa_K2589001:Design

Designed by: John Bechtel   Group: iGEM18_WLC-Milwaukee   (2018-10-10)


Short His J-Protein (trc-promoter)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Ensuring our primers would amplify both our promoter and J-protein while maintaining the His tag was a primary consideration in designing this part so that we could induce protein expression and later purify it using the His-tag.



Source

This part was originally derived from the pETail4 plasmid which provided the template from which the J-Protein was subcloned and a 6 residue His tag attached (using PCR primers). This construct was then cloned into pTrc99a and the trc-promoter from pTrc99a was amplified along with our J-protein via PCR to create a sequence wherein our part was downstream of the trc-promoter. This construct was then cloned into pSB1C3 creating our N-Terminus His tagged J-protein under the control of the trc-promoter.


References