Difference between revisions of "Part:BBa I739001"
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+ | BioBrick [https://parts.igem.org/wiki/index.php?title=Part:BBa I739001 BBa I739001] was ligated upstream of BioBrick [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2170221 BBa_K2170221] (TetO-OmpA) which was then ligated upstream of BioBrick [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2170142 BBa_K2170142] (NanoLuc-''Strep'' Tag-double terminator) to assemble BioBricks [https://parts.igem.org/wiki/index.php?title=Part:BBa_BBa_BBa_K2170141 BBa_K2170141] (TetR-TetO-OmpA) and [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2170053 BBa_K2170053] (TetR-TetO-OmpA-NanoLuc-''Strep'' Tag-double terminator. The functionality of both BioBricks could be confirmed by Nano-Glo assaywhich is why the functionality of [https://parts.igem.org/wiki/index.php?title=Part:BBa I739001 BBa I739001] could be validated as well. | ||
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Revision as of 22:26, 16 October 2016
Constitutive expression cassette for TetR +LVA (J23100.B0034.C0040.B0015)
Part Structure
The Biobrick encodes LVA-tagged TetR (BBa_C0040) under control of the constitutive promoter BBa_J23105 followed by the ribosome binding site BBa_B0034. The transcription of tetR is terminated by the double terminator BBa_B0015.
Mode of Action
TetR binds to the pTet regulator (BBa_R0040) and represses transcription. If the inducer [http://openwetware.org/wiki/ATc anhydrotetracycline (ATc)] is added, TetR action is inhibited and the promoter gets derepressed.
Purpose
This Biobrick was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and belongs to the constitutive part of the system. In the project description, this part is also termed Part 1. The constitutively synthesized TetR interacts with the double promoters BBa_I739102 and BBa_I739106 which are parts of composites BBa_I739015 and BBa_I739010 respectively. A similar (but not used) construct in this context is BBa_Q04400.
Testing
Checked for uniqueness of restriction enzyme cleavage sites:
Eco: ok
Xba: ok
Spe: ok
Pst: ok
Additional characterization of BBa I739001 by [http://2016.igem.org/Team:LMU-TUM_Munich Munich 2016]
BioBrick I739001 BBa I739001 was ligated upstream of BioBrick BBa_K2170221 (TetO-OmpA) which was then ligated upstream of BioBrick BBa_K2170142 (NanoLuc-Strep Tag-double terminator) to assemble BioBricks BBa_K2170141 (TetR-TetO-OmpA) and BBa_K2170053 (TetR-TetO-OmpA-NanoLuc-Strep Tag-double terminator. The functionality of both BioBricks could be confirmed by Nano-Glo assaywhich is why the functionality of I739001 BBa I739001 could be validated as well.
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]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]