Team:KCL UK/Parts

Parts Overview


Our project aim is to create a synthetic protein polymer to coat our spinal scaffold which will allow for strong adhesion of the PCL to the site of injury, as well as provide excellent mechanical strength to spine. Mfp-5 is one of the first proteins secreted by the mussel byssus and its natural role is to adhere to underwater substrates. We have taken the coding sequence of the beta(β) isoform of Pvfp-5 and created a genetic toolbox to enable the formation of a Pvfp-5β polymer in E.Coli. The 2020 KCL iGEM Team have designed a co-expression system to enable the in situ polymerisation of Pvfp-5β. Our parts collection consists of 2 BioBricks and 3 composites. All composites are the original work of KCL iGEM 2020 and we have used the tyrosinase sequence from the iGEM registry. Our two inserts are the Pvfp-5β and Tyrosinase coding sequences and these will form two separate vectors after being cloned into our plasmid backbone. Several design considerations have been made to ensure tight transcriptional control, genetic selectivity, high yield purification and minimal toxicity to the host cell. The two vectors will be expressed in parallel to form a polymer of Pvfp-5β in vivo.
Part Name Part Number Type Description Designer Length
T7 Promoter BBa_K3635004 Basic T7 promoter Coding sequence Emily Blundell 19 bp
Ribosome Binding Site J61101 Basic RBS Sequence John Anderson 26 bp
Lac Operator BBa_K3635003 Basic Lac Operator Sequence Emily Blundell 25 bp
Histidine Tag BBa_K3635005 Basic 7xHis Tag Emily Blundell 21 bp
pBluescript II SK+ f1 Ori BBa_K3635006 Basic Origin of Replication Emily Blundell 306 bp
Ampicillin Resistance BBa_K3635007 Basic AmpR Emily Blundell 966 bp
Pvfp-5β Coding Sequence BBa_K3635000 Basic Pvfp-5β Insert Emily Blundell/Abigail Conner 528 bp
Tyrosinase Coding Sequence BBa_K3033013 Coding Tyrosinase Insert Philip Naderev P.Lagniton 891 bp
Renervate Plasmid Backbone BBa_K3635008 Composite T7 promoter+RBS+lac operator+7xhis-tag+pBluescript II SK+ f1 ori+AmpR Emily Blundell 2219 bp
Pvfp-5β Expression Vector BBa_K3635009 Composite T7 promoter+RBS+lac operator+Pvfp-5β+7xhis-tag+pBluescript II SK+ f1 ori+AmpR Emily Blundell 2755 bp
Tyrosinase Expression Vector BBa_K3635013 Composite T7 promoter+RBS+lac operator+Tyrosinase+7xhis-tag+pBluescript II SK+ f1 ori+AmpR Emily Blundell 3116 bp

Click below to see our parts:

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Pvfp-5β DNA Insert
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Tyrosinase DNA Insert
3 / 4
Renervate Plasmid Backbone
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Pvfp-5β Expression Vector
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Tyrosinase Expression Vector