Composite

Part:BBa_K3762008:Design

Designed by: Andrea Grimsdatter Stallvik, Martin Eide Lien   Group: iGEM21_NTNU-Trondheim   (2021-09-11)


Sulfide sensor


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 211
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 537
    Illegal NheI site found at 560
    Illegal NheI site found at 1980
    Illegal NheI site found at 2003
    Illegal PstI site found at 211
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2279
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 211
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 211
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 865


Design Notes

This composite part was synthesized by IDT as a linear sequence, and cloned into a plasmid vector using Gibson assembly. The success of the cloning was verified through sequencing. It was later discovered that a mistake was made when ordering the sequence, resulting in the start codon for the CreiLov gene (Part:BBa_K3762001) being omitted. This is a likely cause of the part not working as planned.

Source

The genes Psqr, SqrR, and SQR, were sourced from genome of Rhodobacter Capsulatus SB1003 (GenBank accession no. CP001312), and then codon-optimized using the codon optimization tool from IDT, as described on the individual part pages. The gene encoding CreiLov was described in from a paper by Zou et al[1], and found when researching fluorescent proteins that work anaerobically. The other parts in this composite part are existing biobrick parts from the registry.

References

[1] Zou, W., Le, K., & Zastrow, M. L. (2020). Live‐Cell Copper‐Induced Fluorescence Quenching of the Flavin‐Binding Fluorescent Protein CreiLOV. Chembiochem : a European Journal of Chemical Biology, 21(9), 1356–1363. https://doi.org/10.1002/cbic.201900669