Difference between revisions of "Part:BBa K1985017"
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The part was first validated with a diagnostic restriction digest using EcoRI and PstI and agarose gel electrophoresis. The expected band sizes from the digest were: 3140 base pairs for the plasmid backbone and 2129 for the insert. A 1kB plus DNA marker was used to verify the sizes of the bands and it was confirmed that the correct plasmid had been produced. | The part was first validated with a diagnostic restriction digest using EcoRI and PstI and agarose gel electrophoresis. The expected band sizes from the digest were: 3140 base pairs for the plasmid backbone and 2129 for the insert. A 1kB plus DNA marker was used to verify the sizes of the bands and it was confirmed that the correct plasmid had been produced. | ||
| − | [[File:PSB1A3--AraC-pBAD--MamOP gel.png||400px|thumb|centre|Figure 1. Agarose gel of the restriction digest of | + | [[File:PSB1A3--AraC-pBAD--MamOP gel.png||400px|thumb|centre|Figure 1. Agarose gel of the restriction digest of BBa_K1985017 in pSB1A3, with EcoRI and PstI.]] |
Revision as of 10:33, 17 October 2016
AraC pBAD mamOP
This is a composite part of part BBa_K1321333 (Imperial iGem, 2014), BBa_K1985006 and BBa_K1985000 and . It was used in pSB1A3.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 2081 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
Illegal AgeI site found at 1946
Illegal AgeI site found at 2030
Illegal AgeI site found at 2939
Illegal AgeI site found at 3600 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 2561
Illegal BsaI.rc site found at 1232
Illegal SapI site found at 961
Usage and Biology
Usage: The mamO gene was used to initiate the formation of magnetite by "nucleating" the crystal particles, allowing further development. MamP forms part of a proposed electron transport complex (with mamT and X) in vivo, which should form magnetite. The part was used to assemble further composite parts BBa_K1985008 and BBaK1985009 in pSB1A3.
BBa_K1321333 is a regulatory part is made up of the Arabinose-Inducible promoter, pBAD, and its transcriptional inhibitor/activator, AraC. It was used to give increased control over the expression of part BBa_K1985007.
Biology: For more information on the biology of mamO and mamP see parts BBa_K1985006 and BBa_K1985000 and for more information on the AraC pBAD promoter please see BBa_K1321333.
Validation
The part was first validated with a diagnostic restriction digest using EcoRI and PstI and agarose gel electrophoresis. The expected band sizes from the digest were: 3140 base pairs for the plasmid backbone and 2129 for the insert. A 1kB plus DNA marker was used to verify the sizes of the bands and it was confirmed that the correct plasmid had been produced.
