IGEM:IMPERIAL/2006/project/Oscillator/project browser/Full System/Implementation

Super Parts	Not applicable
Actual Part
Sub Parts	Prey Construct	Predator Construct

• Specifications
• Design
• Modelling
• Implementation
• Testing/Validation

Ligation strategy

Ligating the Prey Construct:

For	Stage 1	Stage 2
<bbpart>J37034</bbpart>= (<bbpart>C0261</bbpart>+<bbpart>I13504</bbpart>)
<bbpart>J37015</bbpart>= (<bbpart>F2620</bbpart>+<bbpart>J37034</bbpart>)

Ligating the Polycystronic Predator Construct:

For	Stage 1	Stage 2	Stage 3	Stage 4
<bbpart>J37023</bbpart> (<bbpart>"PCR'd"</bbpart>)
<bbpart>J37024</bbpart>= (<bbpart>J37023</bbpart>+<bbpart>B0014</bbpart>)
<bbpart>J37025</bbpart>= (<bbpart>B0034</bbpart>+<bbpart>J37024</bbpart>)
<bbpart>J37033</bbpart>= (<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)
<bbpart>J37019</bbpart>= (<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)
<bbpart>J37036</bbpart>= (<bbpart>J37019</bbpart>+<bbpart>J37025</bbpart>)

Ligating the Monocystronic Predator Construct:

For	Stage 1	Stage 2	Stage 3	Stage 4	Stage 5
<bbpart>J37023</bbpart> (<bbpart>"PCR'd"</bbpart>)
<bbpart>J37024</bbpart>= (<bbpart>J37023</bbpart>+<bbpart>B0014</bbpart>)
<bbpart>J37025</bbpart>= (<bbpart>B0034</bbpart>+<bbpart>J37024</bbpart>)
<bbpart>J37033</bbpart>= (<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)
<bbpart>J37019</bbpart>= (<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)
<bbpart>J37031</bbpart>= (<bbpart>J37019</bbpart>+<bbpart>B0014</bbpart>)
<bbpart>J37018</bbpart>= (<bbpart>J37031</bbpart>+<bbpart>R0062</bbpart>)
<bbpart>J37035</bbpart>= (<bbpart>J37018</bbpart>+<bbpart>J37025</bbpart>)

Quality control

Part	Logo	Built	Sequenced	Primers Used for Sequencing
<bbpart>J37018</bbpart>		No	N/A	N/A
<bbpart>J37019</bbpart>		Yes	Yes	Forward pSB1(AK3/A3/A2) Sequence: TAT TGT CTC ATG AGC GG Seconday Structure: None
<bbpart>J37023</bbpart>		Yes	Yes	?
<bbpart>J37024</bbpart>		No	N/A	N/A
<bbpart>J37025</bbpart>		No	N/A	N/A
<bbpart>J37031</bbpart>		No	N/A	N/A
<bbpart>J37033</bbpart>		Yes	Not sequenced	N/A
<bbpart>J37035</bbpart>		No	N/A	N/A
<bbpart>J37036</bbpart>		Yes	?	?

WetLab work entries

• The prey has been constructed and tested but is waiting parameter extraction.
• The predator construct has not been ligated.

• Issues included non-functional registry parts such as the registry Aiia and B0015 stop codon.
• We found a new source of AiiA and created J37023 which is a LVA and Flag tagged Aiia gene.