Team:York/Parts


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

So what do all of these parts do?

BBa_K1807000

  • 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.

BBa_K1807001

  • 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.

BBa_K1807002

  • 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).

BBa_K1807003

  • This part is a protein coding device containing the Polyphosphate Kinase (PPK) gene from Candidatus Accumulibacter phosphatis strain BA-91. PPK produces polyphosphate, more specifically PPK catalyzes the reversible conversion of the γ-phosphate of ATP to the end of the polyphosphate chain. This enzyme is responsible for the formation of long chain polyphosphate molecules (up to a thousand orthophosphate residues long).

BBa_K1807004

  • This part is a protein coding device containing the Polyphosphate Kinase (PPK) gene from Candidatus Accumulibacter phosphatis strain Sk-12. PPK produces polyphosphate, more specifically PPK catalyzes the reversible conversion of the γ-phosphate of ATP to the end of the polyphosphate chain. This enzyme is responsible for the formation of long chain polyphosphate molecules (up to a thousand orthophosphate residues long).

BBa_K1807005

  • This part is a protein coding device containing the Polyphosphate Kinase (PPK) gene from Candidatus Accumulibacter phosphatis strain UW-1. PPK produces polyphosphate, more specifically PPK catalyzes the reversible conversion of the γ-phosphate of ATP to the end of the polyphosphate chain. This enzyme is responsible for the formation of long chain polyphosphate molecules (up to a thousand orthophosphate residues long).

BBa_K1807006

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

BBa_K1807007

  • This part is a protein coding device containing the Phosphate Specific Transporter (Pst) gene from Candidatus Accumulibacter phosphatisSCAB. This transporter consists of 4 individual genes- pstA, pstC, pstB and pstS. PstA and pstC are phosphate transporter permeases, pstB is a phosphate transporter ATPase and pstS is a phosphate transporter periplasmic binding protein.