Difference between revisions of "Part:BBa K3866023"

Revision as of 21:07, 25 September 2021

sbm - Methylmalonyl-CoA mutase GB compatible with B2-B5

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

Usage and Biology

Catalyzes the interconversion of succinyl-CoA and methylmalonyl-CoA. Could be part of a pathway that converts succinate to propionate.

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-Sbm (C7 + C8) with: EcoRI, Expected bands: 2518bp, 1733bp, Positive result: C7 + C8

Source

Synthesized by IDT.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1697
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1697
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1697
Illegal BamHI site found at 985
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1697
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1697
Illegal NgoMIV site found at 2012
Illegal AgeI site found at 149
1000
COMPATIBLE WITH RFC[1000]

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

@@ Line 18: / Line 18: @@
 ===Verification of cloning===
-[[File:T--Thessaly--Sbm0--digestion.png|800px|thumb|none|<i><b>Figure 3.</b> (C= Cut, U=Uncut) Restriction digestion of pTrc:RBS (C1 + C2) with: EcoRI + EcoRV, Expected bands: 1280bp, 896bp, Positive result: C1 + C2</i>]]
+[[File:T--Thessaly--Sbm0--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>]]
 ===Source===