Part:BBa_K4179014
ChimeraII under Rhlr promoter + P2A + mCherry reporter
This composite part comprises of an umbelliferone-6 prenyl transferase ChimeraII (BBa_K4179012), which was engineered by the team of Technion 2022-Israel, under the rhlr-regulated promoter (BBa_R0071). Downstream to ChimeraII there is a P2A sequence (BBa_K4179005), which is a self-cleavage element that induces ribosomal skipping during translation of a protein inside the cell. Downstream to which there is an mCherry (BBa_K106005) reporter gene, and an rrnB terminator sequence (BBa_K4047025).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 2152
Illegal PstI site found at 778
Illegal PstI site found at 1792 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1357
Illegal PstI site found at 778
Illegal PstI site found at 1792 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 843
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 2152
Illegal PstI site found at 778
Illegal PstI site found at 1792 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 2152
Illegal PstI site found at 778
Illegal PstI site found at 1792
Illegal NgoMIV site found at 883
Illegal AgeI site found at 1431 - 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
In this genetic system, the mCherry expression is tied directly to the start codon of ChimeraII, while not being covalently bound to it. In such a design the mCherry signal corresponds to a 1:1 ratio, at the very least, which allows for a lower estimate of ChimeraII's expression.
ChimeraII enzyme is the first enzyme in the decursin biosynthesis pathway, which the team was attempting to clone into a bacterial system. For more information about the ChimeraII enzyme, and its construction process, visit its page in the registry (BBa_K4179012), or visit the team’s wiki page.
The team used this sequence in the rhlr-tdpp7-m/cherry plasmid, which already holds the rhlr gene.
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