June

June 23rd

Goal: Design plasmids that express agaA construct

Procedure:

Using Geneious, we first created the agaA construct:

• Promoter BBa_J23108 from the Anderson's promoter collection
• RBS from the RBS Calculator of Salis lab specific for E coli
• BioBrick Prefix + Scar from the iGEM Parts webpage
• agaA sequence codon optimized for E coli 12 (IDT online tool). We checked with Generious that there were no restriction sites for EcoRI, SpeI, ZbaI and PstI.
• Histidine tag: 6 Histidines in C-terminal were added
• Stop codon
• BioBrick scar + BioBrick suffix from the iGEM Parts webpage

We will set the parameters in the machine and run the reaction overnight.

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July

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August

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September

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October

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