Difference between revisions of "Part:BBa K3866023"
 
(4 intermediate revisions by the same user not shown)
Line 4: Line 4:
  
  
−
[[Image:T--Thessaly--sbm0snap.png|700px|thumb|none|<I><b>Figure 1.</b> The level 0 module : pupd2- sbm (illustration from SnapGene)</i>]]
+
[[Image:T--Thessaly--sbm0snap.png|800px|thumb|none|<I><b>Figure 1.</b> The level 0 module : pupd2- sbm (illustration from SnapGene)</i>]]
  
 
===Usage and Biology===
 
===Usage and Biology===
Line 18: Line 18:
 
===Verification of cloning===
 
===Verification of cloning===
  
−
[[File:T--Thessaly--Sbm0--digestion.png|500px|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===
 
===Source===
−
Synthesized by IDT.
+
Synthesized by Twist Biosciences.
 +
 
 +
===Experimental Use and Experience===
 +
This part showed functionality at the following parts: <bbpart>BBa_K3866026</bbpart>, <bbpart>BBa_K3866029</bbpart>,
 +
<bbpart>BBa_K3866031</bbpart>
  
 
===Sequence and Features===
 
===Sequence and Features===

Latest revision as of 22:36, 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 Twist Biosciences.

Experimental Use and Experience

This part showed functionality at the following parts: BBa_K3866026, BBa_K3866029, BBa_K3866031

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