Difference between revisions of "Part:BBa K1045017:Design"
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===Source=== | ===Source=== | ||
− | This composite part was constructed using hybridization oligos for [[Part:BBa_K1045010|BBa_1045010]] and [[Part:BBa_K1045011|BBa_1045011]]. The hybridization oligos were ordered from Sigma-Aldrich. In addition, parts that were obtained either by amplification of plasmid DNA (inverse terminator [[Part:BBa_K1045009|BBa_1045009]]) or ''M. smegmatis'' chromosomal DNA (DarR ORF, [[Part:BBa_K1045001|BBa_1045001]] were employed. The chromosomal DNA of ''Mycobacterium smegmatis'' was purchased from DSMZ ([http://www.dsmz.de/catalogues/details/culture/DSM-43756.html?tx_dsmzresources_pi5%5BreturnPid%5D=304 DSM No. 43756]). Part [[Part:BBa_E0240|BBa_E0240]] derived from the plasmid in the distribution kit 2013. | + | This composite part was constructed using hybridization oligos for [[Part:BBa_K1045010|BBa_1045010]] and [[Part:BBa_K1045011|BBa_1045011]]. The hybridization oligos were ordered from Sigma-Aldrich. In addition, parts that were obtained either by amplification of plasmid DNA from the distribution kit 2013(inverse terminator [[Part:BBa_K1045009|BBa_1045009]]) or ''M. smegmatis'' chromosomal DNA (DarR ORF, [[Part:BBa_K1045001|BBa_1045001]]) were employed. The chromosomal DNA of ''Mycobacterium smegmatis'' was purchased from DSMZ ([http://www.dsmz.de/catalogues/details/culture/DSM-43756.html?tx_dsmzresources_pi5%5BreturnPid%5D=304 DSM No. 43756]). Part [[Part:BBa_E0240|BBa_E0240]] derived from the plasmid in the distribution kit 2013. |
===References=== | ===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 | 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 14:22, 19 October 2013
DarR reporter system
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 806
Illegal NheI site found at 829
Illegal NheI site found at 909
Illegal NheI site found at 932
Illegal NotI site found at 718 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 731
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 860
Illegal AgeI site found at 168 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1630
Design Notes
DarR fused to the terminator was cut out from BBa_K1045016 and ligated in a pre-fixing composition into BBa_K1045015.
Source
This composite part was constructed using hybridization oligos for BBa_1045010 and BBa_1045011. The hybridization oligos were ordered from Sigma-Aldrich. In addition, parts that were obtained either by amplification of plasmid DNA from the distribution kit 2013(inverse terminator BBa_1045009) or M. smegmatis chromosomal DNA (DarR ORF, BBa_1045001) were employed. The chromosomal DNA of Mycobacterium smegmatis was purchased from DSMZ ([http://www.dsmz.de/catalogues/details/culture/DSM-43756.html?tx_dsmzresources_pi5%5BreturnPid%5D=304 DSM No. 43756]). Part BBa_E0240 derived from the plasmid in 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