Part:BBa_K845009:Design
paraBAD_gfp_cyaA
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 500
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 344
Illegal BamHI site found at 1538
Illegal XhoI site found at 494 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 500
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 500
Illegal AgeI site found at 179
Illegal AgeI site found at 484 - 1000INCOMPATIBLE 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