Device
Part:BBa_K1172912:Design
Designed by: Tore Bleckwehl Group: iGEM13_Bielefeld-Germany (2013-09-19)
Part 1 of the Biosystem TetOR alive (pRha TetR alr)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1156
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1080
Illegal BamHI site found at 1782 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1198
Illegal AgeI site found at 1498 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 955
Design Notes
- Amplification of the rhamnose promoter PRha (BBa_K194004) with its pSB1C3 backbone using specific Gibson-Primer:
pRhA_rev (41 bp)
5'-TTGTTTCTCCTCTTTCTAGTATTCATTACGACCAGTCTAAA-3'
pSB1C3_rha_suf (42 bp)
5'-GTGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAG-3'
- Amplification of the repressor TetR with Gibson-Primer containing the RBS (BBa_B0034):
tetR_fwd (41 bp)
5'-TACTAGAAAGAGGAGAAACAAATGTCCAGATTAGATAAAAG-3'
tetR_rev (22 bp)
5'-TTATTAAGCTACTAAAGCGTAG-3'
- Amplification of the Biobrick BBa_K1172903 with Gibson-Primer containing the RBS (BBa_B0034):
alr_tetR_1 (67 bp)
5'GTAGCCGTCAAGTTGTCATAATACTAGAAAGAGGAGAAATACTAGATGCAAGCGGCAACTGTTGTG-3'
alr_tetR_2 (42 bp)
5'-CTGCAGCGGCCGCTACTAGTA-3'
- Ligation via [http://2013.igem.org/Team:Bielefeld-Germany/Labjournal/ProtocolsPrograms#Gibson_assembly Gibson-Assembly].
Source
- Rhamnose promoterBBa_K194004
- TetR repressor BBa_C0040
- Alanine racemase and double terminator BBa_K1172903