Difference between revisions of "Part:BBa K4726000:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | + | In order to clone this genetic construct onto our expression vector (pET-3a) we added NdeI site in 5' and BamHI site in 3' of the gene. We added TATATA sequences before the NdeI site and after the BamHI site to assist with the restriction digestion (those sites are not on the part sequence). The part sequence included a 6x His-tag on the C-terminus of the protein. | |
===Source=== | ===Source=== | ||
| − | + | This gene originates from <i>Flavonifractor plautii</i>, a bacterium from the human gut microbiota. It encodes a N-acetyl-α-D-galactosamine deacetylase. | |
| + | |||
| + | The amino sequence is: | ||
| + | |||
| + | MRNRRKAVSLLTGLLVTAQLFPTAALAADSSESALNKAPGYQDFPAYYSD | ||
| + | SAHADDQVTHPDVVVLEEPWNGYRYWAVYTPNVMRISIYENPSIVASSDG | ||
| + | VHWVEPEGLSNPIEPQPPSTRYHNCDADMVYNAEYDAMMAYWNWADDQGG | ||
| + | GVGAEVRLRISYDGVHWGVPVTYDEMTRVWSKPTSDAERQVADGEDDFIT | ||
| + | AIASPDRYDMLSPTIVYDDFRDVFILWANNTGDVGYQNGQANFVEMRYSD | ||
| + | DGITWGEPVRVNGFLGLDENGQQLAPWHQDVQYVPDLKEFVCISQCFAGR | ||
| + | NPDGSVLHLTTSKDGVNWEQVGTKPLLSPGPDGSWDDFQIYRSSFYYEPG | ||
| + | SSAGDGTMRVWYSALQKDTNNKMVADSSGNLTIQAKSEDDRIWRIGYAEN | ||
| + | SFVEMMRVLLDDPGYTTPALVSGNSLMLSAETTSLPTGDVMKLETSFAPV | ||
| + | DTSDQVVKYTSSDPDVATVDEFGTITGVSVGSARIMAETREGLSDDLEIA | ||
| + | VVENPYTLIPQSNMTATATSVYGGTTEGPASNVLDGNVRTIWHTNYAPKD | ||
| + | ELPQSITVSFDQPYTVGRFVYTPRQNGTNGIISEYELYAIHQDGSKDLVA | ||
| + | SGSDWALDAKDKTVSFAPVEAVGLELKAIAGAGGFGTAAELNVYAYGPIE | ||
| + | PAPVYVPVDDRDASLVFTGAWNSDSNGSFYEGTARYTNEIGASVEFTFVG | ||
| + | TAIRWYGQNDVNFGAAEVYVDGVLAGEVNVYGPAAAQQLLFEADGLAYGK | ||
| + | HTIRIVCVSPVVDFDYFSYVGEHHHHHH | ||
===References=== | ===References=== | ||
| + | [1] Rahfeld P, Sim L, Moon H, Constantinescu I, Morgan-Lang C, Hallam SJ, Kizhakkedathu JN, Withers SG. An enzymatic pathway in the human gut microbiome that converts A to universal O type blood. Nat Microbiol. 2019 Sep;4(9):1475-1485. doi: 10.1038/s41564-019-0469-7. Epub 2019 Jun 10. PMID: 31182795. | ||
Latest revision as of 01:25, 10 October 2023
N-acetyl-α-D-galactosamine deacetylase_Flavonifractor plautii
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 865
Illegal EcoRI site found at 1411
Illegal NgoMIV site found at 1584
Illegal NgoMIV site found at 1894 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
In order to clone this genetic construct onto our expression vector (pET-3a) we added NdeI site in 5' and BamHI site in 3' of the gene. We added TATATA sequences before the NdeI site and after the BamHI site to assist with the restriction digestion (those sites are not on the part sequence). The part sequence included a 6x His-tag on the C-terminus of the protein.
Source
This gene originates from Flavonifractor plautii, a bacterium from the human gut microbiota. It encodes a N-acetyl-α-D-galactosamine deacetylase.
The amino sequence is:
MRNRRKAVSLLTGLLVTAQLFPTAALAADSSESALNKAPGYQDFPAYYSD SAHADDQVTHPDVVVLEEPWNGYRYWAVYTPNVMRISIYENPSIVASSDG VHWVEPEGLSNPIEPQPPSTRYHNCDADMVYNAEYDAMMAYWNWADDQGG GVGAEVRLRISYDGVHWGVPVTYDEMTRVWSKPTSDAERQVADGEDDFIT AIASPDRYDMLSPTIVYDDFRDVFILWANNTGDVGYQNGQANFVEMRYSD DGITWGEPVRVNGFLGLDENGQQLAPWHQDVQYVPDLKEFVCISQCFAGR NPDGSVLHLTTSKDGVNWEQVGTKPLLSPGPDGSWDDFQIYRSSFYYEPG SSAGDGTMRVWYSALQKDTNNKMVADSSGNLTIQAKSEDDRIWRIGYAEN SFVEMMRVLLDDPGYTTPALVSGNSLMLSAETTSLPTGDVMKLETSFAPV DTSDQVVKYTSSDPDVATVDEFGTITGVSVGSARIMAETREGLSDDLEIA VVENPYTLIPQSNMTATATSVYGGTTEGPASNVLDGNVRTIWHTNYAPKD ELPQSITVSFDQPYTVGRFVYTPRQNGTNGIISEYELYAIHQDGSKDLVA SGSDWALDAKDKTVSFAPVEAVGLELKAIAGAGGFGTAAELNVYAYGPIE PAPVYVPVDDRDASLVFTGAWNSDSNGSFYEGTARYTNEIGASVEFTFVG TAIRWYGQNDVNFGAAEVYVDGVLAGEVNVYGPAAAQQLLFEADGLAYGK HTIRIVCVSPVVDFDYFSYVGEHHHHHH
References
[1] Rahfeld P, Sim L, Moon H, Constantinescu I, Morgan-Lang C, Hallam SJ, Kizhakkedathu JN, Withers SG. An enzymatic pathway in the human gut microbiome that converts A to universal O type blood. Nat Microbiol. 2019 Sep;4(9):1475-1485. doi: 10.1038/s41564-019-0469-7. Epub 2019 Jun 10. PMID: 31182795.