Difference between revisions of "Part:BBa K1985007"
Line 2: Line 2:
 
<partinfo>BBa_K1985007 short</partinfo>
 
<partinfo>BBa_K1985007 short</partinfo>
  
−
This is a composite part of Part: mamO
+
This is a composite part of part [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1321333 BBa_K1321333] (Imperial iGem, 2014) and [https://parts.igem.org/Part:BBa_K1985006 BBa_K1985006]. It was used in pSB1A3.
−
 
+
−
<!-- Add more about the biology of this part here
+
−
===Usage and Biology===
+
  
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1985007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1985007 SequenceAndFeatures</partinfo>
 +
 +
 +
===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 mamO, P and X in vivo to form magnetite. The part was used to assemble further composite parts [https://parts.igem.org/Part:BBa_K1985017 BBa_1985017],  [https://parts.igem.org/Part:BBa_K1985008 BBa_K1985008] and [https://parts.igem.org/Part:BBa_K1985009 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 see part [https://parts.igem.org/Part:BBa_K1985006 BBa_K1985006] and for more information on the AraC pBAD promoter please see [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1321333 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: 2029 base pairs for the plasmid backbone and 838 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--MamO gel.jpeg||400px|thumb|centre|Figure 1. Agarose gel of the restriction digest of BBa_K1985007 in pSCB13, with EcoRI and PstI.]]
 +
 +
  
  

Revision as of 09:55, 17 October 2016

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


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
  • 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. It was used in combination with the proposed electron transport complex of mamO, 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.

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: 2029 base pairs for the plasmid backbone and 838 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_K1985007 in pSCB13, with EcoRI and PstI.