All of our BioBricks were assembled according to the RFC[10] or RFC[25] standard. For all basic parts the RFC[10] standard was used. Because the RFC[10] scar codes for a stop codon this assembly standard does not allow the generation of fusion proteins. Therefore all fusion proteins were assembled by RFC[25] standard. Thus all RFC[10] and RFC[25] digestion sites were eliminated from the BioBricks either directly when planing the genesynthesis or by quick-change PCR. Only one of the BioBricks which we supply for the community could not be freed from all digestion sites, because one part of the BioBrick

• GMK_TK90 "Kill-Switch": [http://parts.igem.org/Part:BBa_K404113 BBa_K404113] & [http://parts.igem.org/Part:BBa_K782063 BBa_K782063] were improved as both parts were wrong as they both contained a forbidden PstI restriction site that made it impossible to ligate BioBricks downstream of the BioBrick. A corrected BioBrick was provided ([http://parts.igem.org/Part:BBa_K2170060 BBa_K2170060])

• Tet-Operator: [http://parts.igem.org/Part:BBa_I739001 BBa_I739001] was improved by confirming functionality, adding a Tet-Operator together with an OmpA secretion signal yielding the part [http://parts.igem.org/Part:BBa_K2170141 BBa_K2170141] which can be used by teams for the SEC-dependent secretion in the bacterial periplasm.

• The collection of eukaryotic signal peptides in RFC[25] was expanded significantly by identifying non-working signal peptides ([http://parts.igem.org/Part:BBa_I712009 BBa_I712009], as not in RFC[25]), characterizing working signal peptides ([http://parts.igem.org/Part:BBa_K157001 BBa_K157001]) and designing, constructing and testing two additional signal peptides with significantly higher functionality ([http://parts.igem.org/Part:BBa_K2170214 BBa_K2170214] & [http://parts.igem.org/Part:BBa_K2170215 BBa_K2170215])

Fig.2 Finally it's done. Ready to send our 2016 BioBricks

After month of cloning we could finally pack and send our 65 BioBricks on Friday the 14th of September.