Difference between revisions of "Part:BBa K845009:Design"

(→‎Source)
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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This Biobrick has to be inserted in <i>ΔcyaA</i> bacteria.
  
 
Our safety notes:
 
Our safety notes:

Revision as of 13:57, 30 September 2012

paraBAD_gfp_cyaA


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 500
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 344
    Illegal BamHI site found at 1538
    Illegal XhoI site found at 494
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 500
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 500
    Illegal AgeI site found at 179
    Illegal AgeI site found at 484
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2172
    Illegal BsaI.rc site found at 1169
    Illegal SapI site found at 161
    Illegal SapI.rc site found at 1662
    Illegal SapI.rc site found at 1902


Design Notes

This Biobrick has to be inserted in ΔcyaA bacteria.

Our safety notes:

https://static.igem.org/mediawiki/2012/4/48/CyaA.pdf

https://static.igem.org/mediawiki/2012/2/2a/ParaBAD-gfp.pdf

Source

The part comes from the plasmid collection Alon (http://www.weizmann.ac.il/mcb/UriAlon/Papers/Zaslaver_Ecoli_library.pdf). However, pBAD is a natural promoter in E. Coli. It regulates the production of three proteins (AraA, AraB and AraD) which form the arabinose operon. Their production enables the use of arabinose as a carbon source. It has six regulation sites, five of them are devoted to AraC fixation (three are repressing, one has a dual activity and one is an activator); the last one is a CRP binding site (positive activity). This promoter is activated when both activated CRP and AraC are bind to it. GFP comes from the protein comes from Aequorea victoria. This jellyfish uses GFP (Green Fluorescent Protein) in order to convert the blue luminescence emitted by the aequorine into a green luminescence. Aequorea victoria is a jellyfish that can be found off the coast of north America.

pBAD usage in biology: http://www.univ-orleans.fr/sciences/BIOCHIMIE/L/Illustrations%20cours/SLO-5BC03%20Regulation%20expression%20genome/Procaryotes/SLO-5BC03-COURS3.pdf

References