Part:BBa_K592023

B0032-BFP

Blue Fluorescent Protein with B0032 RBS, this part can be added downstream to any promoter, assembled into pSB3K3 backbone and tested for promoter strength. B0032 is the standard RBS for promoter tests in pSB3K3 backbone.

Contribution

Group: iGEM2016_Hong_Kong_HKUST
Author: iGEM2016_Hong_Kong_HKUST

Summary:
We hypothesized that the expression level of mTagBFP is affected by the presence or absence of a terminator. To check if that was true, we compared fluorescence outputs from BFP generators with terminator, versus one without a terminator.
The results showed that the observed blue fluorescence outputs were lowered when the transcription was not properly terminated.

In this experiment, we have improved BBa_K592023 (BBa_B0032-BBa_K592100) by adding a terminator BBa_B1006 3’ to this part in hoping to provide future users with better reporters for their assays. We also would like to raise the awareness on the standards of parts measurement through this investigation. We cautioned on the direct use of previous measurement data for promoters generated using BBa_K592023, and we suggest future iGEM teams that undertake measurement experiments of any kind not to omit parts that might appear irrelevant in the measurement cassettes.

*Please refer to BBa_K1899001 as the improved part.

Figure 1. Comparison of the expression levels of a BFP generator with and without a terminator.(a) Circuit diagram illustrates experimental setup of the experiment harboring BBa_B0032. (b) Experimental results of the experiment harboring BBa_B0032. Error bars represent SEM of 3 independent experiments on 3 different days.

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
  COMPATIBLE WITH RFC[1000]