Difference between revisions of "Part:BBa K3510006"
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<partinfo>BBa_K3510006 short</partinfo> | <partinfo>BBa_K3510006 short</partinfo> | ||
| − | <b><font color= red>Important: This construct does not include a RBS upstream of the | + | <b><font color= red>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</font color= red></b> |
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 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). | ||
Latest revision as of 12:19, 23 October 2020
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 199
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1401