Composite

Part:BBa_K1899007

Designed by: iGEM2016_HKUST   Group: iGEM16_Hong_Kong_HKUST   (2016-10-10)


phlFp-B0032-Lacl-B0032-TetR-GFP

This construct is designed to estimate the workload of E. coli and measure the fluorescence expression levels by the promoter when it is ligated to the tristable switch.


Results

The fold change between construct A (pSB3K3-BBa_phlFp-E0240) and negative control (pSB3K3-BBa_E0240), and that of construct E (pSB3K3-BBa_phlFp-B0032-C0012-B0032-C0040-E0240) is 13.2 and 1.8 times respectively.

Owing to the increase in number of coding regions, the workload of the cells is highly increased. The production rate of fluorescence should therefore be lower. It is acceptable to obtain a drop in the RFU level between the two constructs.

Fig 1. Comparison on fluorescence expression levels of construct A (BBa_phlFp-E0240) and construct E (BBa_phlFp-B0032-C0012-B0032-C0040-E0240). Negative control represents BBa_E0240. Characterization was done using E. coli strain JW0336. Cells were first precultured overnight and were subcultured to mid-log phase where GFP emission measurements were made using an EnVision® multilabel reader. This result was obtained by combining 3 characterization data obtained in 3 different days. Error bar present SD from 3 biological replicates.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1240
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2629


[edit]
Categories
//chassis/prokaryote/ecoli
Parameters
None