Part:BBa_K5459001

Regulatory protein for copper detection.
This is a part designed for the expression of the CueR protein, utilizing a strong promoter BBa_J23119 to drive its expression for our copper ion detection system.

This part can be used in conjunction with BBa_K5459002, but our experimental results have shown that BBa_K5459002 can also function independently by leveraging the endogenous CueR at the genomic level in E. coli. We have compared the differences between these two scenarios:

We have observed that in a two-plasmid system, the use of the BBa_J23119 promoter to initiate CueR expression indeed impacts the fluorescence levels of the entire system. This influence can be harnessed to develop copper ion detectors with varying performance characteristics.
High throughput promoter experiment

We observed that all promoters in the promoter library exhibit a certain level of promoter activity, indicating their significant role in transcriptional regulation. The initiation strength of the promoters shows a degree of variability, with the lowest induction strength being 5-fold, and the highest reaching up to 10-fold. To gain a deeper understanding of the relationship between the activity of these promoters and their sequence characteristics, we employed Sanger sequencing to determine the nucleotide sequences of these promoters. Subsequently, based on the sequencing results, we conducted a correlation analysis between the initiation strength of the promoters and their sequence features, and built a mathematical model accordingly. The detailed construction process and parameter settings of this model can be consulted on our provided "Model" page. Through this model, we aim to reveal the specific mechanisms by which promoter sequence characteristics affect their initiation activity, providing a theoretical foundation and experimental guidance for subsequent research on gene expression regulation.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]