Part:BBa_K1985007
AraC pBAD mamO
This is a composite part of part BBa_K1321333 (Imperial iGem, 2014) and BBa_K1985006. 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 - 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. It was used in combination with the proposed electron transport complex of mamT, P and X in vivo to form magnetite. The part was used to assemble further composite parts BBa_1985017, 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.
The part was used in pSB1A3 rather than pSB1C3 as it was cotransformed with pec86, a cytochrome maturation factor, which was in a chloramphenicol resistant plasmid.
Biology: For more information on the biology of mamO see part BBa_K1985006 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.
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