Team:UCLouvain/Parts

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 Overview

In our project we worked with multiple plasmids. We tried different reporter plasmids: RFP and GFP in high and low copy plasmids with different antibiotics resistance genes. We worked also with two different conjugative plasmids: pSW23T ans pK18mob.

The plasmids used as reporter :

We worked with an Red Fluorescent Protein constitutive generator in the form of the biobrick Bba_J04450 and with the Green Fluorescent Protein generator in the form of a construction of two biobricks R0010, a LacI dependent promoter, and E0840, a weak RBS promoterless GFP generator.
Both of those construction have been placed in high replicative plasmids backbones: pSB1R3 (with the specific resistance of the backbone R=C, chloramphenicol; A, Ampicillin; K, Kanamycin).
They have also been placed in low replicative plasmids backbones: pSB4R5 (with the specific resistance of the backbone R = C, chloramphenicol; A, Ampicillin; K, Kanamycin).

The submitted parts:

We sent two different biobricks.

• The first one, K2654001, is our GFP generator with a weak RBS and a LacI dependent promoter. We used this part to discern our donor with the green color. We wanted a weak RBS to avoid a deflection of the ribosomes from the vital translations.
• The second one is , K2654002, is a customable sgRNA with a switchable crRNA sequence. This biobrick was used to create two sgRNA’s for txo differnts targets: The RFP Bba_E1010 and the Ampicillin Resistance gene.