Revision as of 21:57, 18 September 2015

Part Table

Source Organism	Gene	Part Name	Function	Characterised	Sequenced	Submitted
Expression vector “pAdapt” in pSB1C3	lacZalpha	BBa_K1807000	Used as cloning vector	✔	✔	✔
Escherichia coli	EcPPX	BBa_K1807001	Phosphate Kinase		✔	✔
Escherichia coli	EcPPK	BBa_K1807002	Phosphate Kinase		✔	✔
Escherichia coli	EcPstSCAB		Phosphate specific transporter		✔
Kingella oralis	KoPPK	BBa_K1807006	PolyPhosphate Kinase		✔	✔
Sinorhizobium meliloti	SmPstSCAB		Phosphate specific transporter	✔	✔	✔
Candidatus Accumulibacter phosphatis	ApPPK BA-91	BBa_K1807003	PolyPhosphate kinase	✔	✔	✔
Candidatus Accumulibacter phosphatis	ApPPK SK-12	BBa_K1807004	PolyPhosphate kinase	✔	✔	✔
Candidatus Accumulibacter phosphatis	ApPPK UW-1	BBa_K1807005	PolyPhosphate kinase	✔	✔	✔
Candidatus Accumulibacter phosphatis	ApPstSCAB	BBa_K1807007	Phosphate specific transporter		✔	✔

This device allows for the IPTG-inducible expression of lacZα peptide which in the presence of Xgal in the medium and chromosomal lacZΔM15 produces blue-coloured colonies. The device was used as an expression vector. SmaI restriction sites flank the lacZ alpha coding sequence. Cutting the part with this enzyme allows for the in-frame insertion of any desired protein coding sequence that contains the splice-in flanking sequences (they can be found in the design section).
BBa_K1807000 was designed as a blue-white screening device that would also be easily used in Gibson Assembly. Surrounding the lacZ alpha coding sequence are two SmaI restriction sites (CCC/GGG). SmaI is a blunt-end endonuclease- we used it to simulatenously linearize our vector and remove the lacZ alpha coding sequence. Our BioBricks contained overhangs that make them compatible with the SmaI-digested BBa_K1807000. The overhang sequences used to make this part are as follows:
- BBa_K1807000 Assembly 5'end Overhang (complementary to pSB1C3): cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGAG
- BBa_K1807000 Assembly 3'end Overhang (complementary to pSB1C3): TACTAGTAGCGGCCGCTGCAGtccggcaaaaaagggcaag The subparts of the device are as follows: BBa_R0011, BBa_B0034, BBa_E0038, BBa_B0015.

This part codes for the exopolyphosphatase (PPX) enzyme of Escherichia coli. PPX is able to release phosphate residues from the ends of a polyphosphate chain.

This part is a protein coding device containing the Polyphosphate Kinase (PPK) gene from Escherichia coli. PPK produces polyphosphate, more specifically PPK catalyzes the reversible conversion of the γ-phosphate of ATP to the end of the polyphosphate chain (Akiyama et al., 1992). This enzyme is responsible for the formation of long chain polyphosphate molecules (up to a thousand orthophosphate residues long).

@@ Line 225: / Line 225: @@
      <h3>BBa_K1807002</h3>
      <ul>
-     <li>This part is a protein coding device containing the Polyphosphate Kinase (PPK) gene from Escherichia coli. PPK produces polyphosphate, more specifically PPK catalyzes the reversible conversion of the γ-phosphate of ATP to the end of the polyphosphate chain. </li>
+     <li>This part is a protein coding device containing the Polyphosphate Kinase (PPK) gene from Escherichia coli. PPK produces polyphosphate, more specifically PPK catalyzes the reversible conversion of the γ-phosphate of ATP to the end of the polyphosphate chain (Akiyama et al., 1992). This enzyme is responsible for the formation of long chain polyphosphate molecules (up to a thousand orthophosphate residues long).</li>
      </ul>
    </div>