Part:BBa_K3762006:Design

Reactive sulfur species sensor

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 214
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 536
  Illegal NheI site found at 559
  Illegal PstI site found at 214
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 491
  Illegal XhoI site found at 835
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 214
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 214
  
• 1000
  COMPATIBLE WITH RFC[1000]

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, and SqrR 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