Question:

Should we separate plasmids from Biobricks? Should each section apply to each construct?

Contents 1 Construction of plasmids 1.1 Assembling DNA parts 1.2 Confirming 1.3 Corroborating sequences 1.4 Conclusions 2 Transforming Synechocystis PCC 6803 2.1 Transformation 2.2 Reestreaking 2.3 Confirming DNA 2.4 Conclusions 3 Revealing phenotypes 3.1 Microscopy 3.2 Luminometer readings 3.3 Conclusions

Construction of plasmids

Our first attempts to construct plasmid pSB1C3_IntK was through a Gibson Assembly reaction of 6 different parts (see construct detail BBa_K743004) all at once, however we were unsuccessful due to many reasons, many which became clear afterwards (see NOTEPAD WEEK SOMETHING ABOUT GIBSON). We did many attempts betting on depletion of PpsbAB (140bp) due to T5 exonuclease where we changed concentration ratios of the parts, nevertheless we were not able to assemble the parts.
During late June we were able to Biobrick the recombination sites ( Part:BBa_K743000 and Part:BBa_K743001) and we decided to use standard assembly to construct a smaller version of pSB1C3_IntK where

Assembling DNA parts

Confirming

Corroborating sequences

Conclusions

Transforming Synechocystis PCC 6803

Transformation

Reestreaking

Confirming DNA

Conclusions

Revealing phenotypes

Microscopy

Luminometer readings

Conclusions