Part:BBa_K1391017:Experience

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Applications of BBa_K1391017

MAV1212-RFP is a backbone vector designed for creation of miRNA sensing circuits. Selection for successful transformation can be done with Ampicillin, as the plasmid carries AmpR.

Colonies expressing the MAV1212-RFP will appear red from expression of RFP. Sites for Gateway Cloning (LR) are around the RFP gene. This means that after a LR reaction, colonies with successful LR can be selected by picking colorless colonies that had the RFP gene replaced by targeted sequences, usually a promoter and an output.

LacZ gene bordered by Golden Gate sites follows the RFP gene. This is intended as a way for inserting miRNA target sites to the gene. Selection for successful addition of target sites to the plasmid can be done by plating colonies on Xgal plates and selecting for colorless colonies. Colonies still expressing LacZ will appear blue.

Sites for Gibson assembly are also present for addition of expression vectors expressing a reporter gene (for flow-cytometry). This can be skipped and transfection reporting done by co-transfecting the reporter gene with the MAV1212 circuit.

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