Part:BBa_K3174002

Blue Fluorescent Protein with Strong Promoter and Strong RBS

This part has the blue fluorescent protein (mTagBFP) gene inserted downstream of a strong promoter and strong ribosome binding site.

BBa_J23104 is part of the Anderson Series of constitutive promoters designed by iGEM2006_Berkeley.

BBa_B0034 is an RBS based on the Elowitz repressilator and was designed in 2003. Its relative strength was characterized by IIT Madras in 2016 and by Team Warsaw in 2010.

BBa_K592100 is a blue fluorescent protein designed by iGEM11_Uppsala-Sweden in 2011. It has a maximum emission of 456 nm.

Fig.3: Differential growth rates of 5 BFP plasmids in TOP10 cells: (left to right) BBa_K3174002, BBa_K3174003, BBa_K3174004, BBa_K3174006, and BBa_K3174007. Single colonies are scarce for BBa_K3174002 and BBa_K3174003 , more abundant for BBa_K3174004, and most abundant for BBa_K3174006 and BBa_K3174007. This differential growth rate is due to the different levels of metabolic burden each strain expresses due to their relative promoter and RBS strengths.

Usage and Biology

UT Austin iGEM 2019 Team transformed this part into the E.coli DH10B burden monitor (https://2019.igem.org/Team:Austin_UTexas) and used it as a standard with which cellular burden could be quantified (by means of a GFP expression vs growth rate model) for other BioBricks that had been similarly transformed into the burden strain.

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]