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Parts Overview

Phactory Parts

Our parts collection consists of four basic parts and one improved composite part. All parts were generated during our project. Some parts relate to RecBCD enzyme & its inhibition and some to the translation detection via mTurquoise.

Parts Related to RecBCD

The RecBCD enzyme, also called Exonuclease V, is both a helicase that unwinds and separates linear DNA strands, and a nuclease that degrades linear DNA1. It consists of three subunits: RecB, RecC and RecD. RecBCD complex is essential for cell viability as it plays an important role in repair of DNA double-strand breaks by initiating homologous recombination2. However, RecBCD enzyme is also a highly processive at destruction of foreign DNA, like phage DNA3.

RecBCD is regulated by a cis-acting DNA sequence known as Chi (Crossover Hotspot Instigator) that activates its recombination-promoting functions. Interaction with Chi causes an attenuation of the RecBCD enzyme's vigorous nuclease activity4. Since Chi binds RecBCD with a greater affinity than other dsDNA substrate, this can be used to inhibit RecBCD activity in TX-TL, to more efficiently express linear DNA in TX-TL.

We provide basic parts containing the full coding region of the subunit RecC (BBa_K2722000) and RecD (BBa_K2722001), which can be used for an independent study of these subunits. Moreover, you can find the corresponding inhibitor Chi6 (BBa_K2722002) in our parts collection.

Parts Related to mTurquoise

The fluorescent reporter protein mTurquoise can be used for a variety of applications. For example, it can be used to measure translation efficiency. Our parts collection includes the coding sequence of mTurquoise (BBa_K2722003) for easy fusion to target proteins or cloning into other functional plasmids, as well as an expression construct of mTurquoise with a constitutive promoter (BBa_K2722004).

The parts are flanked by a BioBrick prefix and suffix and contain pSB1C3 as a backbone and come from risk group 1 organisms.

To learn more about our parts visit the Registry.

iGEM Munich BioBrick List