Difference between revisions of "Part:BBa K5196014:Design"

 
 
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<partinfo>BBa_K5196014 short</partinfo>
 
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<partinfo>BBa_K5196014 SequenceAndFeatures</partinfo>
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__TOC__
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===Design Notes===
 
===Design Notes===
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This primer amplifies ALDH fragment 1 and generates 5' overhangs.
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<p>We considered maintaining the integrity of the T7 promoter site and kanamycin resistance sequence during our construct formation. We decided to use the XbaI and PspXI restriction sites to insert the ALDH sequence [2,3].
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</p>
  
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<p>
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This primer regenerates the 5’ overhang of ALDH Fragment 1.
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</p>
  
 
===Source===
 
===Source===
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<p>
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The source of the ALDH gene is <em>Pseudonocardia dioxanivorans</em> strain CB1190 [1,4]. The pET-28-NicA2 plasmid was derived from <em>E. coli</em>.
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</p>
  
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The ALDH gene is native to P. Putida.
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===References===
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<p>[1] Sales, C. M., Grostern, A., Parales, J. V., Parales, R. E., & Alvarez-Cohen, L. (2013). Oxidation of the Cyclic Ethers 1,4-Dioxane and Tetrahydrofuran by a Monooxygenase in Two Pseudonocardia Species. Applied and Environmental Microbiology, 79(24), 7702-7708. https://doi.org/10.1128/AEM.02418-13
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</p>
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<p>[2] Biolabs, N. E. (n.d.). <em>Nebuilder.</em> NEBuilder. https://nebuilder.neb.com/#!/add/
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</p>
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<p>[3] <em>Integrated DNA Technologies</em>. IDT. (2024, September 11). https://www.idtdna.com/page
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</p>
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<p>[4] Database, A. P. S. (2022, November 1). <em>Alphafold protein structure database.</em> https://alphafold.ebi.ac.uk/entry/Q88CR0
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</p>
  
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===References===
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<p>Sequence and Features:</p>
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<partinfo>BBa_K5196014 SequenceAndFeatures</partinfo>

Latest revision as of 06:39, 2 October 2024


ALDH Gibson Primer 1

Design Notes

We considered maintaining the integrity of the T7 promoter site and kanamycin resistance sequence during our construct formation. We decided to use the XbaI and PspXI restriction sites to insert the ALDH sequence [2,3].

This primer regenerates the 5’ overhang of ALDH Fragment 1.

Source

The source of the ALDH gene is Pseudonocardia dioxanivorans strain CB1190 [1,4]. The pET-28-NicA2 plasmid was derived from E. coli.

References

[1] Sales, C. M., Grostern, A., Parales, J. V., Parales, R. E., & Alvarez-Cohen, L. (2013). Oxidation of the Cyclic Ethers 1,4-Dioxane and Tetrahydrofuran by a Monooxygenase in Two Pseudonocardia Species. Applied and Environmental Microbiology, 79(24), 7702-7708. https://doi.org/10.1128/AEM.02418-13

[2] Biolabs, N. E. (n.d.). Nebuilder. NEBuilder. https://nebuilder.neb.com/#!/add/

[3] Integrated DNA Technologies. IDT. (2024, September 11). https://www.idtdna.com/page

[4] Database, A. P. S. (2022, November 1). Alphafold protein structure database. https://alphafold.ebi.ac.uk/entry/Q88CR0

Sequence and Features:


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]