EXISTING PARTS FROM THE REGISTRY

Existing Parts from the Registry: list

• BBa_K300009/BBa_I4102 - PoPS->3OC6HSL sender device
• BBa_F2620 - 3OC6HSL -> PoPS Receiver
• BBa_J61001 - R6K Origin of replication
• BBa_J23100, BBa_J23101, BBa_J23105, BBa_J23106, BBa_J23110, BBa_J23114, BBa_J23116, BBa_J23118 - constitutive promoters from Anderson's collection
• BBa_P1004 - chloramphenicol resistance cassette
• BBa_K125500 - GFP fusion brick
• BBa_J72008 - phi80 integration helper plasmid pInt80-649

<partinfo>BBa_K300009</partinfo>/<partinfo>BBa_I4102</partinfo> - PoPS->3OC6HSL sender device

<partinfo>BBa_F2620</partinfo> - 3OC6HSL -> PoPS Receiver

<partinfo>BBa_J61001</partinfo> - R6K Origin of replication

<partinfo>BBa_J23100</partinfo>, <partinfo>BBa_J23101</partinfo>, <partinfo>BBa_J23105</partinfo>, <partinfo>BBa_J23106</partinfo>, <partinfo>BBa_J23110</partinfo>, <partinfo>BBa_J23114</partinfo>, <partinfo>BBa_J23116</partinfo>, <partinfo>BBa_J23118</partinfo> - constitutive promoters from Anderson's collection

<partinfo>BBa_P1004</partinfo> - chloramphenicol resistance cassette

<partinfo>BBa_K125500</partinfo> - GFP fusion brick

This part can be useful to construct fluorescent fusion proteins. It is composed by a tail domain (the GFP <partinfo>K125500</partinfo>) with a transcriptional terminator (<partinfo>BBa_B0015</partinfo>) downstream.

Other protein domains can be fused upstream of this part in order to create chimeric fluorescent proteins, or it can be ligated to tags useful for low-cost protein purification.

This part was used do design the following BioBrick measurement systems:

• <partinfo>BBa_K300086</partinfo>
• <partinfo>BBa_K300088</partinfo>
• <partinfo>BBa_K300090</partinfo>
• <partinfo>BBa_K300091</partinfo>
• <partinfo>BBa_K300092</partinfo>
• <partinfo>BBa_K300099</partinfo>

to test these contructs:

• <partinfo>BBa_K300002</partinfo>
• <partinfo>BBa_K300093</partinfo>
• <partinfo>BBa_K300094</partinfo>
• <partinfo>BBa_K300095</partinfo>
• <partinfo>BBa_K300084</partinfo>
• <partinfo>BBa_K300097</partinfo>

respectively. All of the tested parts are synthetic fusion tags whose activity could be measured by assembling a promoter with RBS upstream and a tail domain with terminator downstream, thus yielding the measurement systems. <partinfo>BBa_K300005</partinfo> was used as a tail domain with terminator downstream. It has been assembled to test the correct folding of the resulting fusion protein by measuring the GFP and to test the affinity tag performance with a proof of concept protein.

In all of the measurement parts, GFP could be successfully detected in bacteria harbouring the measurement parts in high copy plasmids. In this condition, GFP was detected by using an excitation filter at 485nm and an emission filter at 540nm in a Infinite F200 microplate reader (Tecan).

Raw GFP curve	Mean (dGFP/dt)/O.D. over the exponential phase (under the hypothesis that GFP half-life in fusion contructs is similar to the original one - <partinfo>BBa_E0040</partinfo>)
Raw GFP curve	Mean (dGFP/dt)/O.D. over the exponential phase (under the hypothesis that GFP half-life in fusion contructs is similar to the original one - <partinfo>BBa_E0040</partinfo>)

<partinfo>BBa_J72008</partinfo> - phi80 integration helper plasmid pInt80-649