Composite

Part:BBa_K316027

Designed by: IC 2010 Team   Group: iGEM10_Imperial_College_London   (2010-10-26)
Revision as of 10:32, 26 October 2010 by Ks907 (Talk | contribs)


B. subtilis transformation vector with LacI, targets amyE locus.

This vector has been designed using the amyE 5' and 3' integration sequences for integration into B.subtilis genome


AmyE locus This vector has been designed using the amyE 5' BBa_K143008 and 3 BBa_K143009' integration sequences for integration into B. subtilis genome. Insertion into the amyE locus provides a selection marker as the bacterium will no longer be able to breakdown starch. An iodine assay can be used to confirm integration. This phenotype makes the transformed bacterium considerably less likely to survive in natural environments.


Chloramphenicol Resistance This vector also contains a positive selection marker, flanked by two dif sites. Chloramphenicol acetyltransferase BBa_J31005 provides resistance to chloramphenicol antibiotic. Dif BBa_K316002 sites allow excision of the antibiotic marker after integration, thus allowing the same marker to be used again or as a precaution against horizontal gene transfer.


Blunt end cloning site PmeI restriction site BBa_K316013 is designed as a cloning site. Due to the 8bp recognition sequence it is a rare site that can be used to cut the vector only once.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2922
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1141


For more information about our project please visit our wiki [http://2010.igem.org/Team:Imperial_College_London] or take the tour [http://2010.igem.org/Team:Imperial_College_London/Tour/Page_One] to learn more about the project.


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