Part:BBa_K2248002
PArsRGFP
This composite part is made of up a SalI Restriction Enzyme Site, E. coli-derived Arsenic Regulator promoter (PArsR), Ribosome Binding Site, Green Fluorescent Protein reporter (GFP), Double Terminator. This composite part is combined with another composite part that has the same Parts except that the GFP reporter is replaced with Arsenic Regulator gene.
PArsR promoter sequence has a binding site for ArsR. ArsR is a negative repressor and will release its transcriptional regulation in the presence of arsenic ions. If ArsR is not present, then GFP is not regulated. If ArsR is present, then the GFP reporter should turn on and fluoresce only in the presence of arsenic.
Usage and Biology
This plasmid was incubated with reagents from the Promega E. coli S30 Extract System for circular DNA. This allows for the in vitro transcription/translation of the gene coding sequences of a plasmid. Testing BBa_K2248002 against a NoDNA negative control and BBa_J364001 positive control allowed us to see that GFP was being made from BBa_K2248002. We can see here that in the absence of ArsR, GFP is produced from BBa_K2248002. More work will need to be done to know whether ArsR will repress GFP expression.
Figure 1: GFP Expression of BBa_K2248002 in vitro using Light Cycler 480. Plasmid DNA was incubated with the reagents needed to transcribe and translate GFP and compared against negative (No DNA) and positive controls (BBa_J364001).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 756
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