Difference between revisions of "Part:BBa K3866024"
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<partinfo>BBa_K3866024 short</partinfo>
 
<partinfo>BBa_K3866024 short</partinfo>
  
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[[Image:T--Thessaly--YgfG0snap.png|800px|thumb|none|<I><b>Figure 1.</b> The level 0 module : pupd2- ygfG (illustration from SnapGene)</i>]]
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[[Image:T--Thessaly--YgfG0snap.png|800px|thumb|none|<I><b>Figure 1.</b> The level 0 module : pupd2- sbm (illustration from SnapGene)</i>]]
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===Usage and Biology===
 
===Usage and Biology===
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===Verification of cloning===
 
===Verification of cloning===
  
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[[File:T--Thessaly--YgfG0--digestion.png|800px|thumb|none|<i><b>Figure 3.</b> (C= Cut, U=Uncut) Restriction digestion of pUPD2-Sbm (C7 + C8) with: EcoRI, Expected bands: 2518bp, 1733bp, Positive result: C7 + C8</i>]]
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[[File:T--Thessaly--YgfG0--digestion.png|600px|thumb|none|<i><b>Figure 3.</b> (C=Cut, U=Uncut) Restriction digestion of pUPD2-YgfG (C1 + C2) with: EcoRV + HindIII, Expected bands: 1718bp, 1174bp, Positive result: C1 + C2</i>]]
  
 
===Source===
 
===Source===
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Synthesized by IDT.
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Synthesized by Twist Biosciences.
  
 
===Sequence and Features===
 
===Sequence and Features===

Latest revision as of 22:12, 25 September 2021


ygfG - Methylmalonyl-CoA decarboxylase GB compatible with B2-B5

Figure 1. The level 0 module : pupd2- ygfG (illustration from SnapGene)

Usage and Biology

Catalyzes the decarboxylation of (R)-methylmalonyl-CoA to propionyl-CoA. Could be part of a pathway that converts succinate to propanoate.

Design Notes

The sequence was domesticated. We removed BsmBI, BsaI, BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for Golden Braid cloning. This has position B2-B5.

Figure 1.The overhangs of this part in the GoldenBraid Grammar

Verification of cloning

Figure 3. (C=Cut, U=Uncut) Restriction digestion of pUPD2-YgfG (C1 + C2) with: EcoRV + HindIII, Expected bands: 1718bp, 1174bp, Positive result: C1 + C2

Source

Synthesized by Twist Biosciences.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 173
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 632

References

Haller T, Buckel T, Rétey J, Gerlt JA (2000). Discovering newenzymes and metabolic pathways: conversion of succinate to propionate by Escherichia coli. Biochemistry, 39:4622–4629. https://doi.org/10.1021/bi992888d