General part design using RFC[10] & RFC[25]

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

Featured parts

[http://parts.igem.org/wiki/index.php?title=Part:BBa_K2170001 BBa_K2170001] Biotin binding receptor with eMA

Our BioBrick [http://parts.igem.org/wiki/index.php?title=Part:BBa_K2170001 BBa_K2170001] contains an eucaryotic receptor construct with enhanced monomeric avidin, which, after expression in an eucaryotic cell, imparts the binding of biotin on the cell surface. This constucts makes it possible to link cells over a biotinylated protein linker and by this makes the printing of tissue without scaffold even possible.

[http://parts.igem.org/wiki/index.php?title=Part:BBa_K2170002 BBa_K2170002] Biotin binding receptor with scAvidin

Our BioBrick [http://parts.igem.org/wiki/index.php?title=Part:BBa_K2170002 BBa_K2170002] contain an eucaryotic biotin binding receptor with the tetrameric biotin binding protein single chain avidin. As it's sibling BioBrick [http://parts.igem.org/wiki/index.php?title=Part:BBa_K2170001 BBa_K2170001] it imparts the polymerization of cells, after transfection, because of the ability to bind biotinylated protein linkers.

Parts we improved

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])

Impressions from the packaging

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.

Sendbox

<groupparts>iGEM2016 LMU-TUM_Munich</groupparts>