Difference between revisions of "Part:BBa K5101001:Design"

 
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===Source===
 
===Source===
  
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This plasmid, pET29a-pJ23119-RBS-AMP-T7, is constructed using the pET29a vector as the backbone, which is widely used for high-level expression in <i>E. coli</i>. The <i>J23119</i> promoter, sourced from the iGEM Registry of Standard Biological Parts, is a strong, constitutive promoter that ensures robust expression of downstream genes(<partinfo>BBa_J23119</partinfo>). The antimicrobial peptide (AMP) gene included in this plasmid was derived from Lactobacilli in the human vaginal flora(<partinfo>BBa K5101000</partinfo>). This AMP was discovered and initially characterized by our host lab at Nanjing Tech University. The sequence screening was performed in-house, and detailed sequence data is pending publication. To align with scientific documentation standards and for comparison, a similar sequence can be referenced in the NCBI database under Reference Sequence: WP_049159833.1.
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This plasmid, pET29a-p<i>J23119</i>-RBS-AMP-T7, is constructed using the pET29a vector as the backbone, which is widely used for high-level expression in <i>E. coli</i>. The <i>J23119</i> promoter, sourced from the iGEM Registry of Standard Biological Parts, is a strong, constitutive promoter that ensures robust expression of downstream genes(<partinfo>BBa_J23119</partinfo>). The antimicrobial peptide (AMP) gene included in this plasmid was derived from <i>Lactobacillus</i> spp. from the human vaginal flora(<partinfo>BBa_K5101000</partinfo>). This AMP-expressing <i>Lactobacillus</i> spp. was discovered and initially characterized by our host lab at Nanjing Tech University. The detailed sequence data is pending publication. To align with scientific documentation standards and for comparison, a same sequence of the AMP can be referenced in the NCBI database under Reference Sequence: WP_049159833.1.
  
 
===References===
 
===References===

Latest revision as of 07:37, 11 September 2024


Plasmid pET29a-pJ23119-RBS-AMP-T7


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 293
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 263
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

During the design phase, special attention was given to ensure the stability of the AMP gene in the plasmid backbone and the compatibility of the promoter with the E. coli expression system. The RBS was optimized for the E. coli ribosome.


Source

This plasmid, pET29a-pJ23119-RBS-AMP-T7, is constructed using the pET29a vector as the backbone, which is widely used for high-level expression in E. coli. The J23119 promoter, sourced from the iGEM Registry of Standard Biological Parts, is a strong, constitutive promoter that ensures robust expression of downstream genes(BBa_J23119). The antimicrobial peptide (AMP) gene included in this plasmid was derived from Lactobacillus spp. from the human vaginal flora(BBa_K5101000). This AMP-expressing Lactobacillus spp. was discovered and initially characterized by our host lab at Nanjing Tech University. The detailed sequence data is pending publication. To align with scientific documentation standards and for comparison, a same sequence of the AMP can be referenced in the NCBI database under Reference Sequence: WP_049159833.1.

References