Difference between revisions of "Team:York/Parts"
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<br>- BBa_K1807000 Assembly 5'end Overhang (complementary to pSB1C3): cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGAG | <br>- BBa_K1807000 Assembly 5'end Overhang (complementary to pSB1C3): cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGAG | ||
<br>- BBa_K1807000 Assembly 3'end Overhang (complementary to pSB1C3): TACTAGTAGCGGCCGCTGCAGtccggcaaaaaagggcaag | <br>- 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.</li> | + | The subparts of the device are as follows: BBa_R0011, BBa_B0034, BBa_E0038, BBa_B0015.</li> |
</ul> | </ul> | ||
</div> | </div> | ||
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<h3>BBa_K1807001</h3> | <h3>BBa_K1807001</h3> | ||
<ul> | <ul> | ||
− | <li>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.</li> | + | <li>This part codes for the exopolyphosphatase (PPX) enzyme of <i>Escherichia coli</i>. PPX is able to release phosphate residues from the ends of a polyphosphate chain.</li> |
</ul> | </ul> | ||
</div> | </div> | ||
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<h3>BBa_K1807002</h3> | <h3>BBa_K1807002</h3> | ||
<ul> | <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 (Akiyama et al., 1992). This enzyme is responsible for the formation of long chain polyphosphate molecules (up to a thousand orthophosphate residues long).</li> | + | <li>This part is a protein coding device containing the Polyphosphate Kinase (PPK) gene from <i>Escherichia coli</i>. 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> | </ul> | ||
</div> | </div> |
Latest revision as of 23:16, 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 | ✔ | ✔ |
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.