Composite

Part:BBa_K3152019

Designed by: Xixi Song   Group: iGEM19_ASTWS-China   (2019-10-17)


mecI-mec operator-mCherry

This part contains “Anderson Medium-RBS-mecI-mec operator-RBS-mCherry” (BBa_K3152004 ,BBa_K3152005).

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
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 379
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

Then we used pSB1C3-GFP as the backbone and instead GFP of mec operator-mCherry (BBa_K3152005). Then the mecI pathway (BBa_K3152004 ) was added into the plasmid pSB1C3-mec-mCherry. As shown in Figure1 below, bacteria colonies did not present the green fluorescence when our DNA fragments were added to the whole plasmid. Also, because of the present of mecI repressor, the color of mCherry disappeared. Three colonies were chosen from the medium and the plasmids were extracted and verified by digesting with restriction enzyme EcoRI and PstI.

Figure 1 Photos of transformants of “pSB1C3-mecI-mec operator-mCherry” on LB agar broth.


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