Composite

Part:BBa_K3510006

Designed by: team work   Group: iGEM20_Tuebingen   (2020-10-21)


Anderson-Promoter-B12-Riboswitch-Tet-Inverter-System-GFP-Terminator

Important: This construct does not include a RBS upstream of the GFP sequence. When building on our design please make sure you add it in order to allow for translation

This composite part is a Vitamin B12 inducible expression system of a superfolder GFP (BBa_I746916). The included part BBa_K1913008 contains a constitutive Promoter (BBa_J23100) and a riboswitch. Upon binding of Vitamin B12, the riboswitch prevents translation of the following gene. To express GFP in the presence of Vitamin B12, a Tet inverter system (BBa_Q04400) is placed in between. When the encoded repressor is not translated anymore, the GFP is expressed again. Transcriptional termination occurs through the activity of the reliable double terminator (BBa_B0015).

This part was inspired by Team Wageningen 2016, who provided this construct with mRFP (BBa_K1913011). By changing the reporter protein to superfold GFP, we seeked to provide an additional option for future iGEM teams of which the expression can be measured reliably using the fluorescein-based iGEM fluorescence calibration protocol.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 199
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1401


[edit]
Categories
Parameters
None