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Parts



On this page, we present the different DNA parts that shaped our project. Synthetic biology is built on bricks, genetic elements, all well documented, that can be replicated and used by scientists world-wide. These elements are easy to find for model organisms. This is not the case for the chassis of our biostimulant, P. fluorescens. This is why we are happy to provide the community with a list of the parts that we built for this promising organism. We hope they will be useful for future iGEM projects and synthetic biologists all over the world.



Introduction

The wild-type strain Pseudomonas fluorescens SBW25 does not grow on creatinine

All biobricks were designed on Benchling and most of them were synthesized by IDT, which allowed us to codon-optimize them for Pseudomonas putida, the closest host organism to our P. fluorescens. Other parts were derived from the genome of P. fluorescens SBW25 via PCR amplification. All parts were cloned into the indicated vector using In-fusion Assembly.



Best part

Type

Purpose

Link

Name

Representation

Importance in BioMoon

Length

Status

RFC

Composite Operon for creatinine metabolization in P. fluorescens BBa_K5108009 creA-crnA *** 2058 Worked 10


Best part

Type

Purpose

Link

Name

Representation

Importance in BioMoon

Length

Status

RFC

Basic RBS from Pseudomonas protegens BBa_K5108006 aprA CHA0 * 24 Worked 10,1000
Basic RBS from Pseudomonas protegens BBa_K5108007 hcnA CHA0 * 25 Not used 10,1000
Basic RBS from Pseudomonas aeruginosa BBa_K5108008 hcnA PAO1 * 25 Not used 10,1000
Basic Creatinase to convert creatine into sarcosine BBa_K5108003 creA ** 1212 Worked 10
Basic Creatinine amidohydrolase to convert creatinine intro creatine BBa_K5108004 crnA ** 809 Worked 10
Basic Chaperone protein involved in the regulations of mRNA in P. fluorescens BBa_K5108004 crnA ** 809 Worked 10
Basic Catalase that decomposes H2O2 into water and oxygen from P. fluorescens BBa_K5108002 katB * 1542 Successful cloning 10