Part:BBa_K3202022

msfGFP

We introduced this new part to the iGEM parts, and have characterized this part in several of our composite parts. This part is used to demonstrate the function of our bistable system.

Characterization

The newly introduced msfGFP is very sensitive according to our results. Comparing to BN006/Contains BBa_K3202029, this new part shows significant difference in both AraC-pBAD inducer system and Xyls-Pm inducer system. Qualitative and quantitative results are shown below.

Figure 1. The left centrifuge tube contains BN006(backbone), and the one on the right contains BN072(AraC-pBAD-msfGFP)

Figure 2. The left centrifuge tube contains BN006(backbone), and the one on the right contains BN073(Xyls-Pm-msfGFP)

Figure 1. The left centrifuge tube contains BN006(backbone), and the one on the right contains BN072(AraC-pBAD-msfGFP)

Figure 2. The left centrifuge tube contains BN006(backbone), and the one on the right contains BN073(Xyls-Pm-msfGFP)

Also, comparing to sfGFP, this new part shows significant difference in both AraC-pBAD inducer system and Xyls-Pm inducer system as well. Quantitative results are shown below.

Figure 3. BN007(AraC-pBAD-sfGFP) and BN072(AraC-pBAD-msfGFP), noticeable difference is shown.

Figure 4. BN009(Xyls-Pm-sfGFP) and BN073(Xyls-Pm-msfGFP), noticeable difference is shown.

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 13
  

References

【1】Aiba H (2007) Mechanism of RNA silencing by Hfq-binding small RNAs. Current Opinion in Microbiology

【2】Anthony LC, Suzuki H, Filutowicz M (2004) Tightly regulated vectors for the cloning and expression of toxic genes. Journal of Microbiological Methods 58: 243-250

【3】Aparicio T, de Lorenzo V, Martínez-García E (2017) Broadening the SEVA Plasmid Repertoire to Facilitate Genomic Editing of Gram-Negative Bacteria. In Hydrocarbon and Lipid Microbiology Protocols: Genetic, Genomic and System Analyses of Pure Cultures

【4】McGenity TJ, Timmis KN, Nogales B (eds), pp 9-27. Berlin, Heidelberg: Springer Berlin Heidelberg Balzer S, Kucharova V, Megerle J, Lale R, Brautaset T, Valla S (2013) A comparative analysis of the properties of regulated promoter systems commonly used for recombinant gene expression in Escherichia coli. Microbial cell factories

【5】Bervoets I, Charlier D (2019) Diversity, versatility and complexity of bacterial gene regulation mechanisms: opportunities and drawbacks for applications in synthetic biology.

【6】Lopez-Walle B, Gauthier M, Chaillet N (2008) Principle of a submerged freeze gripper for micro-assembly. IEEE Trans Robotics 24(4):897–902

【7】Rakotondrabe M, Haddab Y, Lutz P (2009) Development, modelling and control of micro/nano positionning 2 dof stick-slip device, IEEE/ASME Trans. Mechatronics 14(6):733–745

【8】M. Rakotondrabe Y. Haddab and P. Lutz, "Design, development and experiments of a high stroke-precision 2DoF (linear-angular) microsystem" in Proc. 2006 IEEE Int. Conf. on Robotics and Automation: pp. 669 - 674.