Part:BBa_K3425022

iGEM Type IIS standard Level 2 Dummy (MTU-DY)

Type IIS iGEM standard allows for fast and efficient construction of transcriptional units (TU) and multi-transcriptional units (MTU) in groups of four components. These components are needed in Level 1 assemblies (promoter, RBS, CDS and terminator) but might not be needed in higher level assemblies if the amount of transcriptional units in your system is not a multiple of four.

For this reason, placeholders called dummy parts were created based on an existing Modular Assembly method [1]. These parts mimic a basic Type IIS part or a transcriptional unit and can be included in the assembly whenever needed.

This page corresponds to MTU-DY which was cloned to all pEven plasmids (pSB3C1#) and mimics multi-transcriptional units or basic parts. Not present in the part would be the SapI recognition sites, which are positioned so that the fusion sites are cut in the proper way. The part with these sites can be found in Team Uppsala 2020's Parts page. Sequencing results of this part cloned in all pEven plasmids can be found in the Sequence Analysis section and the trace files are in the iGEM Type IIS standard collection page as well as our Parts page.

More information about this cloning standard and dummy parts can be found in Team Uppsala 2020's iGEM Type IIS standard guidebook.

This part is part of iGEM Uppsala 2020's iGEM Type IIS standard collection.

Usage and Biology

Experiments with cloning together all four MTU-DY mimicking four MTUs in pSB1K02 showed that most clones contain deletions or insertions of various sizes in the area of the "multi-dummy" sequence.

This is probably due to the short and repetitive sequence generated by the dummies, since all of them contain the same fusion sites and spacer in between. More information can be found in the composite part BBa_K3425065.

Dummy parts are only supposed to be fillers that allow the cloning reaction to ligate properly when you have less than four parts to assemble. In this sense, this dummy sequence works as intended. However, it is not ideal to have unpredictable mutations occurring systematically. Longer dummy parts with different sequences from each other were designed in silico as a better alternative.

The following table details the part numbers and names of these new dummies (which contain the plasmid they should be cloned into), along with the multi transcriptional unit they are mimicking.

Registry Number	Name	Mimics
BBa_K3425027	pSB3C11-DY	MTU1
BBa_K3425028	pSB3C12-DY	MTU2
BBa_K3425029	pSB3C13-DY	MTU3
BBa_K3425030	pSB3C14-DY	MTU4

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

1. Binder, A., Lambert, J., Morbitzer, R., Popp, C., Ott, T., Lahaye, T., and Parniske, M. (2014) A Modular Plasmid Assembly Kit for Multigene Expression, Gene Silencing and Silencing Rescue in Plants. PLOS ONE. 9, e88218