Day 1

Preparation of Stock Solutions • gBlocks 3 of our gBlocks from IDT had arrived: pCopA MymT HH (pc), MymT sfGFP HH (mg) and TAT Csp1 HH (tc). These arrived in the form of a solid, white powder. We resuspended the gBlocks in different volumes of MilliQ, depending on the weight delivered, to give a stock solution of 10 ng/l. Part name Mass (ng) Volume MilliQ added (l) TAT Csp1 HH 500 50 MymT sfGFP HH 1000 100 pCopA MymT HH 1000 100 • Primers The forward and reverse primers from IDT came as 32.4 nmol and 23.8 nmol of solid respectively. Forward: 5’-CGACTTGATCACGTAGAATTC-3’ Reverse: 5’-ACACGATCGATATAACTGCAGC-3’ For our stock solutions, the primers were suspended in different volumes of MilliQ to give 100M. Primer Amount (nmol) Volume MilliQ added (l) Forward 32.4 324 Reverse 23.8 238 Preparation of reaction solutions • gBlocks Following resuspension, 10l of each DNA solution was added to 90l MilliQ (1:10 dilution) in an Eppendorf to give a final concentration of 1 ng/l and final solution volume of 100l. • Primers Following resuspension, 10l of each primer solution was added to 90l MilliQ (1:10 dilution) in an Eppendorf to give a final concentration of 10M and final solution volume of 100l.

Day 2

Day 3

Day 4

Day 5

Week 2