Coding

Part:BBa_K3286207

Designed by: Ben Kuipers   Group: iGEM19_Wageningen_UR   (2019-10-20)


RpfCch (TMD) sfGFP fusion

This part is similar to part BBa_K3286206, but in this strategy, only the transmembrane domain (TMD) was used to fuse with sfGFP. So again, this part was constructed to test whether part BBa_K3286200 was getting transcribed, translated and transported to the membrane.

Figure 1: Transformation of this part in E. coli Dh5-Alpha cells incubated on a LB agar plate. Fluorescence is visualized by a fluorescence microscope.
Figure 2: Transformation of this part in E. coli Dh5-Alpha cells incubated on a LB agar plate. Fluorescence is visualized by a fluorescence microscope.
Figure 3: Transformation of an E. coli Dh5-Alpha strain producing GFP, incubated on a LB agar plate. Fluorescence is visualized by a fluorescence microscope.


Figure 1 and 2 are showing the observed fluorescence by this part. What can be observed is that cells are actually showing a higher intensity of fluorescence at the edges, especially compared to figure 3 which serves as a positive control. Indicating that this part (but also BBa_K3286200) is getting transcribed, translated and transported to the membrane.


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 227
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 661
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None