Part:BBa_K5415001
Triple HRE biosensor with GFP for low hypoxia detection.
The composite part we have developed is a hypoxia biosensor designed to detect low hypoxic conditions, built in silico using Benchling and HRE gene sequences imported from the https://doi.org/10.1016/j.bbrc.2006.06.043, pMC3-yeGFP imported from Addgene (Plasmid #176181). Gibson and in vivo cloning methods were employed. The constructs were transformed into E. coli.
The biosensor construct features three key components: three hypoxia response elements (HREs), restriction sites (NcoI), and a green fluorescent protein (GFP). The hypoxia response elements are short DNA sequences that respond to the binding of hypoxia-inducible factor (HIF) complexes, which are activated in low oxygen environments. These binded HRE sequences trigger the expression of the GFP, allowing the cell to fluoresce under hypoxic conditions. The use of two restriction sites, NcoI, facilitates further manipulation of the construct if needed.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1
Illegal XhoI site found at 21
Illegal XhoI site found at 35
Illegal XhoI site found at 55
Illegal XhoI site found at 69
Illegal XhoI site found at 89 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 365
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