Difference between revisions of "Part:BBa K1045013:Design"
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+ | Lei Zhang, Weihui Li, and Zheng-Guo He (2013) “DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in Mycobacterium smegmatis”, THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 288, NO. 5, pp. 3085–3096 |
Revision as of 17:38, 4 October 2013
Promoter - DarR operator - GFP generator BBa_E0240
- 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]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 728
Design Notes
For cloning of BBa_K1045013, first, oligos for BBa_K1045012 were hybridized. The hybridization oligos were designed such that their double-stranded hybridization product would correspond to the sequence of BBa_K1045012 cut with EcoRI and SpeI at the prefix and suffix sites. Second, this hybridization product was ligated in front of the GFP generator BBa_E0240 previously cut with EcoRI and XbaI.
Source
The hybridization oligos mentioned under "Design Notes" were purchased from Sigma-Aldrich. The sequence originated from the Parts Registry page of BBa_J23110 and the sequence of the DarR operator found upstream of DarR (Zhang et al., 2013). The plasmid of part BBa_E0240 derived from the distribution kit 2013.
References
Lei Zhang, Weihui Li, and Zheng-Guo He (2013) “DarR, a TetR-like Transcriptional Factor, Is a Cyclic Di-AMP-responsive Repressor in Mycobacterium smegmatis”, THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 288, NO. 5, pp. 3085–3096