Part:BBa_K3889130

Spacer Cassette for Terminator check in Bacillus subtilis

This circuit consists of a dual reporter system in order to check terminator efficiency in Bacillus subtilis.

Usage and Biology

• Measuring efficiency:

The experiment is divided into two cassettes: one reference and a test cassette containing a terminator whose efficiency needs to be determined, as shown by Gale et al.[1].

Fig 1. Spacer Cassette for Terminator check

Fig 2. Spacer replaced by terminator to be checked

The reference(Fig 1) and the test cassette (Fig 2) provide the expression levels of both the fluorescent proteins which could be compared to tell us how efficiently the terminator is working.

Formulae for terminator efficiency [1]:

where

mCherry₀ → mCherry produced by device without terminator

sfGFP₀ → sfGFP produced by device without terminator

Using the device without any changes, TE_Device can be calculated which gives the expression of mCherry in the absence of a terminator.

where

mCherry → mCherry produced by the device with terminator

sfGFP → sfGFP produced by the device with terminator

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 121
  

References

1. Gale, G. A. R., Wang, B., & McCormick, A. J. (2021). Evaluation and Comparison of the Efficiency of Transcription Terminators in Different Cyanobacterial Species. Frontiers in Microbiology, 11. https://doi.org/10.3389/fmicb.2020.624011