Team:CCA San Diego/Parts

Overview

Our part collection documents the constructs used in order to express scl2, a bacterial collagen in S. cerevisiae. Expressing bacterial proteins in a eukaryotic chassis is not common, but possible with a careful design and selection of promoter-terminator pairs and provides the added benefit of a “cut-and-paste” gap-repair cloning design. A modified pRS4II16 backbone was used with a synthetic TEF-based promoter (cpTEF_6-I) with an upstream activating sequence (UASFEC), and strong terminator (PRM9). The three protein domains of scl2 were documented as well as the gene as a composite part and our whole construct. For future researchers interested in manipulating our collagen template, we suggest reading through our Collagene Handbook and Literature Collective on our Contributions page.

Basic Parts

Part Number	Description	Length (bp)
BBa_K3956000	Upstream Activating Sequence FEC	30
BBa_K3956001	30bp_spacer_yeast	30
BBa_K3956003	cpTEF_6-I core promoter	120
BBa_K3956004	5'-UTR	33
BBa_K3956005	Scl2 V Domain (globular)	243
BBa_K3956006	Scl2 enzyme cleaving domain	21
BBa_K3956007	Scl2 triple helix bridging sequence	18
BBa_K3956008	Scl2 CL Domain	696
BBa_K3956009	PRM9 Terminator	243
BBa_K3956016	Shortened pRS4116	3754

Composite Parts

Part Number	Description	Length (bp)
BBa_K3956010	scl2	978
BBa_K3956014	cpTEF_6-I scl2 PRM9	1390