Team:ETHZ Basel/Achievements/BioBrick Toolbox

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BioBrick Toolbox

BioBrick assembly A-parts are used for N-Terminal and C-parts for C-terminal fusion. B-part is the linker that connects the two entities.

To construct the aimed fusion proteins, we first blunt-end cloned our BioBricks into the storage vector pSEVA132 <partinfo>BBa_K422002</partinfo> (Victor de Lorenzo's lab, KanR, pBBR1 ori). All parts are sequence verified and compatible with BBF RFC28 (http://dspace.mit.edu/handle/1721.1/46721)

table 1: Our BrickBox parts for the Chemotaxis BrickBox, the Anchor BrickBox, the Light Sensing BrickBox and the Reporter BrickBox. * compatible for insertion of microRNA binding sites
IDInsertOrientationIDInsertOrientationDescription
<partinfo>BBa_K422003</partinfo>CheBA-part<partinfo>BBa_K422003</partinfo>CheBC-partDemthylase: Demethylates methyl accepting chemotaxis proteins MCPs
<partinfo>BBa_K422005</partinfo>CheRA-part<partinfo>BBa_K422003</partinfo>CheRC-partMethyltransferase: Methylates methyl accepting chemotaxis proteins MCPs
<partinfo>BBa_K422007</partinfo>CheYA-part<partinfo>BBa_K422003</partinfo>CheYC-partInduces tumbling by interacting with the flagellar switch protein FliM
<partinfo>BBa_K422009</partinfo>tetRA-part Tetracyclin repressor: Binds to its operator tetO for spatial localization of a fusion protein to the DNA
<partinfo>BBa_K422010</partinfo>trigA-part Ribosome binding domain of the trigger factor: Binds to the large subunit of the ribosome
<partinfo>BBa_K422011</partinfo>MreBC-partProkaryotic actin homologue: Assembles into helical filaments underneath the cytoplasmic membrane
<partinfo>BBa_K422012</partinfo>Pif3C-partPhytochrome interacting factor: Binds Prf form of PhyB and rapidly dissociates in response to reconversion to the Pr state
<partinfo>BBa_K422013</partinfo>PhyBC-partPhytochromes type II: Biologically inactive form Pr absorbs red light and active configuration Pfr far-red light
<partinfo>BBa_K422014</partinfo>mcyPetA-part CFP variant

We also gave a lot of thoughts how to make BBF RFC28 (method for combinatorial multi-part assembly based on the Type II restriction enzyme AarI) compatible with Tom Knight's OAS standard. Check out our general scheme for the easy construction of Tom Knight's OAS and BBF RFC28 compatible working vectors [1]!

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