Difference between revisions of "Part:BBa K4726001:Design"

 
 
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===Design Notes===
 
===Design Notes===
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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.
  
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===Source===
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This gene was originally taken from the <i>Flavonifractor plautii</i> bacteria. It encodes a α-D-galactosamine galactosaminidase.
  
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The amino acids sequence encoded by this gene is:
  
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===Source===
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MRGKKFISLTLSTMLCLQLLPTASFAAAPATDTGNAGLIAEGDYAIAGNG
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VRVTYDADGQTITLYRTEGSGLIQMSKPSPLGGPVIGGQEVQDFSHISCD
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VEQSTSGVMGSGQRMTITSQSMSTGLIRTYVLETSDIEEGVVYTATSYEA
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GASDVEVSWFIGSVYELYGAEDRIWSYNGGGEGPMHYYDTLQKIDLTDSG
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KFSRENKQDDTAASIPVSDIYIADGGITVGDASATRREVHTPVQETSDSA
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QVSIGWPGKVIAAGSVIEIGESFAVVHPGDYYNGLRGYKNAMDHLGVIMP
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APGDIPDSSYDLRWESWGWGFNWTIDLIIGKLDELQAAGVKQITLDDGWY
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TNAGDWALNPEKFPNGASDALRLTDAIHEHGMTALLWWRPCDGGIDSILY
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QQHPEYFVMDADGRPARLPTPGGGTNPSLGYALCPMADGAIASQVDFVNR
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AMNDWGFDGFKGDYVWSMPECYNPAHNHASPEESTEKQSEIYRVSYEAMV
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ANDPNVFNLLCNCGTPQDYYSLPYMTQIATADPTSVDQTRRRVKAYKALM
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GDYFPVTADHNNIWYPSAVGTGSVLIEKRDLSGTAKEEYEKWLGIADTVQ
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LQKGRFIGDLYSYGFDPYETYVVEKDGVMYYAFYKDGSKYSPTGYPDIEL
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KGLDPNKMYRIVDYVNDRVVATNLMGDNAVFNTRFSDYLLVKAVEISEPD
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PEPVDPDYGFTSVDDRDEALIYTGTWHDDNNASFSEGTARYTNSTDASVV
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FSFTGTSIRWYGQRDTNFGTAEVYLDDELKTTVDANGAAEAGVCLFEALD
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LPAAEHTIKIVCKSGVIDIDRFAYEAATLEPIYEKVDALSDRITYVGNWE
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EYHNSEFYMGNAMRTDEAGAYAELTFRGTAVRLYAEMSFNFGTADVYLDG
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ELVENIILYGQEATGQLMFERTGLEEGEHTIRLVQNAWNINLDYISYLPE
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QDQPTPPETTVTVDAMDAQLVYTGVWNDDYHDVFQEGTARYASSAGASVE
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FEFTGSEIRWYGQNDSNFGVASVYIDNEFVQQVNVNGAAAVGKLLFQKAD
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LPAGSHTIRIVCDTPVIDLDYLTYTTNAHHHHHH
  
 
===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 23:59, 9 October 2023


α-D-galactosamine galactosaminidase_Flavonifractor plautii


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2998
    Illegal PstI site found at 77
    Illegal PstI site found at 571
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2998
    Illegal PstI site found at 77
    Illegal PstI site found at 571
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2998
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2998
    Illegal PstI site found at 77
    Illegal PstI site found at 571
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2998
    Illegal PstI site found at 77
    Illegal PstI site found at 571
    Illegal NgoMIV site found at 84
    Illegal NgoMIV site found at 448
    Illegal NgoMIV site found at 2403
    Illegal NgoMIV site found at 3153
    Illegal AgeI site found at 2106
  • 1000
    COMPATIBLE 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.

Source

This gene was originally taken from the Flavonifractor plautii bacteria. It encodes a α-D-galactosamine galactosaminidase.

The amino acids sequence encoded by this gene is:

MRGKKFISLTLSTMLCLQLLPTASFAAAPATDTGNAGLIAEGDYAIAGNG VRVTYDADGQTITLYRTEGSGLIQMSKPSPLGGPVIGGQEVQDFSHISCD VEQSTSGVMGSGQRMTITSQSMSTGLIRTYVLETSDIEEGVVYTATSYEA GASDVEVSWFIGSVYELYGAEDRIWSYNGGGEGPMHYYDTLQKIDLTDSG KFSRENKQDDTAASIPVSDIYIADGGITVGDASATRREVHTPVQETSDSA QVSIGWPGKVIAAGSVIEIGESFAVVHPGDYYNGLRGYKNAMDHLGVIMP APGDIPDSSYDLRWESWGWGFNWTIDLIIGKLDELQAAGVKQITLDDGWY TNAGDWALNPEKFPNGASDALRLTDAIHEHGMTALLWWRPCDGGIDSILY QQHPEYFVMDADGRPARLPTPGGGTNPSLGYALCPMADGAIASQVDFVNR AMNDWGFDGFKGDYVWSMPECYNPAHNHASPEESTEKQSEIYRVSYEAMV ANDPNVFNLLCNCGTPQDYYSLPYMTQIATADPTSVDQTRRRVKAYKALM GDYFPVTADHNNIWYPSAVGTGSVLIEKRDLSGTAKEEYEKWLGIADTVQ LQKGRFIGDLYSYGFDPYETYVVEKDGVMYYAFYKDGSKYSPTGYPDIEL KGLDPNKMYRIVDYVNDRVVATNLMGDNAVFNTRFSDYLLVKAVEISEPD PEPVDPDYGFTSVDDRDEALIYTGTWHDDNNASFSEGTARYTNSTDASVV FSFTGTSIRWYGQRDTNFGTAEVYLDDELKTTVDANGAAEAGVCLFEALD LPAAEHTIKIVCKSGVIDIDRFAYEAATLEPIYEKVDALSDRITYVGNWE EYHNSEFYMGNAMRTDEAGAYAELTFRGTAVRLYAEMSFNFGTADVYLDG ELVENIILYGQEATGQLMFERTGLEEGEHTIRLVQNAWNINLDYISYLPE QDQPTPPETTVTVDAMDAQLVYTGVWNDDYHDVFQEGTARYASSAGASVE FEFTGSEIRWYGQNDSNFGVASVYIDNEFVQQVNVNGAAAVGKLLFQKAD LPAGSHTIRIVCDTPVIDLDYLTYTTNAHHHHHH

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.