Coding

Part:BBa_K3866024

Designed by: Pericles Vasileiou   Group: iGEM21_Thessaly   (2021-09-24)
Revision as of 21:21, 25 September 2021 by Periclesv (Talk | contribs) (→‎Verification of cloning)


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 IDT.

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

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Categories
Parameters
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