Regulatory

Part:BBa_K4632008:Design

Designed by: Haolong Lai   Group: iGEM23_SCAU-China   (2023-10-09)


Promoter(nirB-medium)


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

We initially attempted to construct a plasmid by connecting the nirBpromoter to the pet-28b plasmid's EGFP gene, but faced repeated failures likely due to the short nirB promoter fragment. We then tried the Ωpcr method without success. Ultimately, we successfully built the plasmid by connecting nirB to the target segment and replaced EGFP with the Gm resistance gene for more reliable characterization in different oxygen conditions.

Source

We obtain the sequence from the literature：Reza N, Akbari Eidgahi M R. Construction of a Synthetically Engineered nirB Promoter for Expression of Recombinant Protein in Escherichia coli[J]. JUNDISHAPUR J MICROB, 2014,7(6).The nirB segment was ordered and synthesized from GUANGZHOU IGE BIOTECHNOLOGY LTD.

References

Reza N, Akbari Eidgahi M R. Construction of a Synthetically Engineered nirB Promoter for Expression of Recombinant Protein in Escherichia coli[J]. JUNDISHAPUR J MICROB, 2014,7(6).