N-terminal linker + double CBD

Double cellulose binding domain (dCBD) using two cellulose binding domains from Trichoderma reesei cellobiohydrolases, with an N-terminal linker and internal linker sequence between the two domains which are derived from the endogenous cellobiohydrolase linker sequence. This part is in RFC(25) Freiberg fusion format to allow for easy use in protein fusions. dCBD binding affinity for cellulose was characterised using sfGFP fusions.

CBDcenA + Linker

Cellulose binding domain (CBD) of Endoglucanase A (cenA) from Cellulomonas fimi with an endogenous C-terminal linker. The part is in Freiburg format (RFC 25) for ease of use in protein fusions.

CBDcipA with N and C-terminal linker

This CBD is from the Cellulosomal -scaffolding protein A (cipA) of Clostridium thermocellum including the endogenous linker sequences at the N and C-terminus. It is in RFC25 format to allow for easy use in protein fusions. At present the cloning for these constructs is still in progress to correct an illegal EcoRI site which was identified in the parts with this CBD. This can be achieved with a silent mutation via site-directed mutagenesis and we aim to send these parts to the registry once this is complete.

Synthetic Phytochelatin (PC) EC20

A general heavy metal-binding peptide consisting of 20 Glu-Cys (EC) repeats in Freiburg format (RFC[25]) to allow for easy use in fusion proteins. We created a library of CBD fusions with this part.

Vitreoscilla haemoglobin (VHb)

This part is a haemoglobin isolated from Vitreoscilla (VHb) which improves the metabolic function of obligate aerobes and facultative anaerobes in low-oxygen conditions. We used this part in G. xylinus to try and improve growth rate and cellulose production.

pSEVA331 and pSEVA321 backbones

These are two broad host range vector backbones for which we have demonstrated use in E. coli and Gluconacetobacter xylinus .

Cellulose synthase operon AcsAB and AcsCD

Designed and refactored from G. xylinus, this operon was inserted into E.coli for cellulose synthesis.

Gluconacetobacter xylinus strains ATCC53582 and Kombucha Isolate

We created and characterised a library of parts for this organism, with the aim to increase the accessibility the chassis for future work in iGEM. By sharing these strains and their genomes on the registry we hope this will contribute to their ease of use and characterisation.