Difference between revisions of "Part:BBa K1985017"

 
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<partinfo>BBa_K1985017 short</partinfo>
 
<partinfo>BBa_K1985017 short</partinfo>
  
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AraC pBAD mamOP
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This is a composite part of parts [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1321333 BBa_K1321333] (Imperial iGem, 2014), [https://parts.igem.org/Part:BBa_K1985006 BBa_K1985006] and [https://parts.igem.org/Part:BBa_K1985000 BBa_K1985000] and . It was used in pSB1A3.
  
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===Usage and Biology===
 
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1985017 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1985017 SequenceAndFeatures</partinfo>
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===Usage and Biology===
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Usage:
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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 [https://parts.igem.org/Part:BBa_K1985008 BBa_K1985008] and [https://parts.igem.org/Part:BBa_K1985009 BBaK1985009] in pSB1A3.
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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.
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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.
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Biology:
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For more information on the biology of mamO and mamP see parts [https://parts.igem.org/Part:BBa_K1985006 BBa_K1985006] and [https://parts.igem.org/Part:BBa_K1985000 BBa_K1985000] and for more information on the AraC pBAD promoter please see [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1321333 BBa_K1321333].
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===Validation===
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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.
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[[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.]]
  
  

Latest revision as of 10:44, 17 October 2016

AraC pBAD mamOP

This is a composite part of parts BBa_K1321333 (Imperial iGem, 2014), BBa_K1985006 and BBa_K1985000 and . It was used in pSB1A3.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 2081
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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.

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 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.

Figure 1. Agarose gel of the restriction digest of BBa_K1985017 in pSB1A3, with EcoRI and PstI.