Composite

Part:BBa_K3745041

Designed by: Jianfei Song   Group: iGEM21_BNDS_China   (2021-09-30)


Prha-Amp with EC6015/Bujard RBS

This part is a rhamnose-induced genetic circuit that contains a Rhamnose promoter, a modified RBS with suitable strength, an ORF of GFP for characterization, and a terminator. It is used for the characterization of an RBS with more suitable strength (BBa_K3745005) for our biosensor.


Usage and Biology

Ailurus biotechnology helped us to adjust the strength of RBS. It is fused on a plasmid with GFP and its strength is measured by detecting the fluorescence intensity of the culture. the Characterization condition is listed below.

With RBS that has modified strength, this part ensures the rhamnose yield is within the dynamic range of the rhamnose-inducible genetic circuit. This will enable the expression of antibiotics resistance gene to significantly change with the concentration of rhamnose (not characterized).

Characterization

The basic Characterization information of this part is listed below:

• Throughput: 2500

• Organism: E. coli

• Strain type: BL21(DE3)

• Temperature: 30°C

• Media: 250 ml LB + 0.1g/L rhamnose (w./v.)

• Period of time: 12 hours

• Plasmid backbone: pSB1C3

Under this condition, the expression of antibiotics resistance gene significantly changes with the concentration of rhamnose.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 161
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 161
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 161
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 161
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 161
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 837


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