Team:Wageningen UR/Parts

Wageningen UR iGEM 2016

 

 

BioBricks overview

Below, you can find all the biobricks we created!

BioBrick number BioBrick name Designer
BBa_K1913025 Wild type plac-FixK2 hybrid promoter best basic part Tianhe
BBa_K1913011 Vitamin b12 riboswitch + mRFP best composite part Carina
BBa_K1913008 vitamin b12 riboswitch Carina
BBa_K1913009 Guanine riboswitch Carina
BBa_K1913010 tetR QPI + mRFP Carina
BBa_K1913012 Guanine riboswitch + guanine riboswitch Carina
BBa_K1913005 lux quorum sensing system + GFP reporter Thomas
BBa_K1913006 434- and lambda cI balance operon + mRFP reporter Thomas
BBa_K1913007 434- and lambda cI operon for tuning protein balance Thomas
BBa_K1913014 3-oxo-hexanoyl-HSL GFP reporter Thomas
BBa_K1913016 434- and lambda cI balance RFP reporter Thomas
BBa_K1913000 chiA for Varroa destructor Lisa
BBa_K1913001 chiB for Varroa destructor Lisa
BBa_K1913002 chiA device regulated by pBAD Lisa
BBa_K1913003 chiB device regulated by pBAD Lisa
BBa_K1913019 Guanine riboswitch BS-yxjA Tianhe
BBa_K1913020 mRFP with degredation tag Tianhe
BBa_K1913021 sGFP with defredation tag Tianhe
BBa_K1913022 Synthetic plac-FixK2 hybrid promoter with RBS Tianhe
BBa_K1913023 Synthetic plac-FixK2 hybrid promoter with RBS Tianhe
BBa_K1913024 Synthetic ptet-FixK2 hybrid promoter with RBS Tianhe
BBa_K1913026 Wild type ptet-FixK2 hybrid promoter Tianhe
BBa_K1913027 Wild type plac-FixK2 hybrid promoter with mRFP Tianhe
BBa_K1913028 Wild type ptet-FixK2 hybrid promoter with mRFP Tianhe
BBa_K1913029 Synthetic plac-FixK2 hybrid promoter +RBS with mRFP Tianhe
BBa_K1913030 Synthetic plac-FixK2 hybrid promoter+RBS with mRFP Tianhe
BBa_K1913031 Synthetic ptet-FixK2 hybrid promoter+RBS with mRFP Tianhe
BBa_K1913032 Toggle Switch device Tianhe
BBa_K1913033 Toggle Switch device Tianhe
BBa_K1913034 Blue light sensor generator Tianhe

Besides, we submitted one part that cannot be classified as a biobrick because it has some illegal restriction sites and it is in the pSB1A3 backbone:

BioBrick number BioBrick name Designer
BBa_K1913015 Cry3Aa with araC/pBAD Jaccoline/Linea

Initally, part BBa_K1913015 was made only to for testing the in vitro assay, jacco, please put the rest of your excuse here ;).

No Cas9 biobrick?!
When we started making the constructs for the Cas9 kill switch, two approaches were taken: one was taking the Cas9 that is available in the iGEM registry (BBa_K1218011) as a starting point for making mutations and expressing Cas9. The other approach was starting with pdCas9 (Addgene plasmid # 46569). While cloning, it proved to be difficult to transfer BBa_K1218011 to another backbone (see also the notebook). Furthermore, when cultures transformed with BBa_K1218011 were checked for Cas9 expression with SDS-PAGE, no convincing Cas9 band could be observed. Because of time limitations we decided to continue working with the Addgene construct for making the mutations, and chose an established system for protein expression. For that reason, no Cas9-biobricks were submitted. Furthermore, the pEVOL construct containing an aminoacyl-synthetase and a tRNA for introducing BipA in response to the TAG stopcodon were isolated from a strain kindly received from George Church (described in Mandel et al., 2015). The MTA that was signed to receive the strain does not allow for redistribution.