Part:BBa_M50041:Design
Positive feedback cusR producer (mutated)
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 53
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 53
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 53
Illegal AgeI site found at 243 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The second half of our genetic construct was “EColi_Promoter_Cassette,” containing strong RBS, the sequence for green fluorescent protein(GFP) with a terminator immediately following it, an ampicillin resistance marker, and a high copy number origin of replication (ORI). “EColi_Promoter_Cassette” was obtained from Dr. Kara Rogers.
We, in fact, designed 2 constructs, BBa_M50039 and BBa_M50041, where the only difference between them was the cusR-inducible promoter sequence. We explored 2 possible sequences of the cusR-inducible promoter: BBa_M50039’s cusR-inducible promoter sequence was from Zahid et al., 4 and BBa_M50041’s promoter sequence comprised the 250 basepairs preceding the cusR gene and was sourced from NCBI.
We ordered the constructs from DNA 2.0. Notably, we were unable to receive BBa_M50041 exactly as prescribed due to a deletion mutation at the final base, which could cause transcription that is unable to be terminated. 
Second figure shows the response to cusR activation by copper ion.
Source
http://biocyc.org/gene?orgid=ECOLI&id=G6319
References
“cusR response regulator in two-component regulatory system with cusS [Escherichia coli str. K-12 substr. MG1655”. (2016, Sep. 10). NCBI Reference Sequences.
Zahid N, Zulfiqar S, Shakoori AR. (2012, March). Functional analysis of cus operon promoter of Klebsiella pneumoniae using E. coli lacZ assay. Gene, 495(1):81-88.
Loftin IR, et al. (2005, July). A novel copper-binding fold for the periplasmic copper resistance protein cusF. Biochemistry, 44(31):10533-10540.