Parts

CONSTRUCT 1: Part BBa_K1163103

This part was created by the Evry 2013 Team. It is iron sensitive with the pAceB promoter region (BBa_K1163101), which contains the FUR binding site and an RBS so another was not needed. FUR (Ferric Uptake Regulator) is a protein which dimerises in the presence of Fe2+ and binds to this promoter to inhibit downstream transcription. In this part the coding region is for the LacI protein (BBa_K1163003). The construct has two terminators (BBa_B0010 and BBa_B0012) because the second is a bad terminator.

CONSTRUCT 2: Part BBa_K2245001

This part is sensitive to LacI with the LacI regulated promoter (BBa_R0010), in the presence of LacI downstream transcription is inhibited. The RBS (BBa_B0034) was chosen because of this part's similarity to the Evry 2013 team's. The coding region is for the amilCP chromoprotein (BBa_K592009) which emits a blue colour. The terminator is BBa_B0010. Together these first two plasmids form the constructs for our final project, the LacI coding region and promoter acting as an inverter system because the iron regulated promoter causes less transcription in the presence of more iron but we needed it to have the opposite effect.

CONSTRUCT 3: Part BBa_K2245000

We created this construct to test the parts without the inverter system, using the same BioBricks as Part BBa_K1163103 but with the amilCP chromoprotein coding region instead of the LacI coding region. This acted as a control to ensure that the inverter system so we could see the effect of the inverter system.

Backbones

We ordered our DNA from IDT so it arrived in backbones with ampicillin resistance. In order to add them to the registry and co-transform them we digested and ligated them into backbones with chloramphenicol resistance (pSB1C3).

List of Parts

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Name	Type	Description	Designer	Length
<a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2245000">BBa_K2245000</a>	Composite	Iron Regulated amilCP	Nikita Shukan	1112
<a class="noul_link part_link" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K2245001">BBa_K2245001</a>	Composite	LacI promoter with amilCP	Nikita Shukan	983

Team:Judd UK/Pages/Parts

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