Difference between revisions of "Part:BBa K4275012:Design"

(→‎Design Notes)
(→‎Design Notes)
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1. A 6x His affinity purification tag(HHHHHH) added to N-terminal following the start codon, to allow for Ni-NTA purification.  
 
1. A 6x His affinity purification tag(HHHHHH) added to N-terminal following the start codon, to allow for Ni-NTA purification.  
  
−
2. The number of cohesins is cut from 9 to 2, and the number of CBM is cut from 2 to 1 to reduce the metabolic burden and probability of successful expression in the E.Coli BL21 recombinant protein expression system.
+
2. The number of cohesins is cut from 9 to 2, and the number of CBM is cut from 2 to 1 to reduce the metabolic burden and increase the probability of successful expression in the E.Coli BL21 recombinant protein expression system.
  
 
3. DNA sequence is codon-optimized based on the codon-usage table of <i>E.coli</i> Strain K12.MG1655.
 
3. DNA sequence is codon-optimized based on the codon-usage table of <i>E.coli</i> Strain K12.MG1655.

Revision as of 08:04, 12 October 2022


CipA1B2C


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 314
    Illegal XhoI site found at 1237
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 543
    Illegal AgeI site found at 325
    Illegal AgeI site found at 669
    Illegal AgeI site found at 868
    Illegal AgeI site found at 1539
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. A 6x His affinity purification tag(HHHHHH) added to N-terminal following the start codon, to allow for Ni-NTA purification.

2. The number of cohesins is cut from 9 to 2, and the number of CBM is cut from 2 to 1 to reduce the metabolic burden and increase the probability of successful expression in the E.Coli BL21 recombinant protein expression system.

3. DNA sequence is codon-optimized based on the codon-usage table of E.coli Strain K12.MG1655.

Source

Clostridium thermocellum

References

1. Anandharaj, Marimuthu et al. "Constructing A Yeast To Express The Largest Cellulosome Complex On The Cell Surface". Proceedings Of The National Academy Of Sciences, vol 117, no. 5, 2020, pp. 2385-2394. Proceedings Of The National Academy Of Sciences, https://doi.org/10.1073/pnas.1916529117.