Difference between revisions of "Part:BBa K4342000"

 
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<partinfo>BBa_K4342000 short</partinfo>
 
<partinfo>BBa_K4342000 short</partinfo>
  
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Confers kanamycin resistance and AZT susceptibility. Designed to ligate to Gene Upstream Homology (Type 1) and Gene Downstream Homology (Type 5) via BsaI digestion for ADP1 Genetic Engineering. Used to ligate to P. destructans Target Sequence Upstream Homology (Type 1) and CymR Repressor (Type 56) via BsaI digestion in P. destructans Detector.
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<h1>Usage and Biology</h1>
  
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The <i> tdk/kan </i> cassette is a highly versatile sequence used for efficiently and effectively engineering the <i> Acinetobacter baylyi </i> ADP1 genome. This part contains the <i> kanR </i> and <i> tdk </i> genes which allow for the selection and counterselection of genetically engineered ADP1 cells. When this cassette is integrated into the ADP1 genome, the <i> kanR </i> gene confers resistance to kanamycin and the <i> tdk </i> gene is lethal to the cell when Azidothymidine is present (Metzgar et. al - 2004).
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===Usage and Biology===
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The 2022 UT Austin iGEM team used the <i> tdk/kan </i> cassette to successfully delete a number of genes, insert DNA sequences into the ADP1 chromosome, and engineer an ADP1-based biosensor for detecting any DNA sequence of interest. The composite parts below utilized the <i> tdk/kan </i> cassette for genetic engineering.
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<span class='h3bb'>Sequence and Features</span>
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<ul>
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<partinfo>BBa_K4342000 SequenceAndFeatures</partinfo>
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<li> <i> ACIAD2049 tdk/kan </i> Cassette </li>
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<li> <i> ACIAD2049 </i> Rescue Cassette</li>
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<li> <i> AcrB tdk/kan </i> Cassette </li>
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<li> <i> AcrB </i> Rescue Cassette </li>
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<li> <i> PbpG tdk/kan Cassette </i> </li>
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<li> <i> PbpG </i> Rescue Cassette </li>
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<li> <i> RecJ tdk/kan </i> Cassette </li>
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<li> <i> RecJ </i> Rescue Cassette </li>
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<li> Broken <i> nptII </i> Gene </li>
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<li> <i> nptII </i> Gene Detector Construct</li>
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<li> <i> TEM-1 </i> Broken Gene </li>
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<li> <i> TEM-1 </i> Gene Detector Construct</li>
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<li> <i> P. destructans </i> Detector Construct</li>
  
  
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===Functional Parameters===
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<h1>Design</h1>
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<partinfo>BBa_K4342000 parameters</partinfo>
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As seen in figure x, the tdk/kan cassette is flanked by bsaI restriction sites on both its 5’ and 3’ ends. These restriction sites produce sticky ends which can ligate to any DNA sequence possessing the necessary restriction sites with the complementary overhangs. Figure X shows how the tdk/kan cassette can be ligated to DNA sequences homologous to ADP1’s genome.
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Revision as of 16:40, 10 October 2022


tdk/kan Cassette

Usage and Biology

The tdk/kan cassette is a highly versatile sequence used for efficiently and effectively engineering the Acinetobacter baylyi ADP1 genome. This part contains the kanR and tdk genes which allow for the selection and counterselection of genetically engineered ADP1 cells. When this cassette is integrated into the ADP1 genome, the kanR gene confers resistance to kanamycin and the tdk gene is lethal to the cell when Azidothymidine is present (Metzgar et. al - 2004).

The 2022 UT Austin iGEM team used the tdk/kan cassette to successfully delete a number of genes, insert DNA sequences into the ADP1 chromosome, and engineer an ADP1-based biosensor for detecting any DNA sequence of interest. The composite parts below utilized the tdk/kan cassette for genetic engineering.

  • ACIAD2049 tdk/kan Cassette
  • ACIAD2049 Rescue Cassette
  • AcrB tdk/kan Cassette
  • AcrB Rescue Cassette
  • PbpG tdk/kan Cassette
  • PbpG Rescue Cassette
  • RecJ tdk/kan Cassette
  • RecJ Rescue Cassette
  • Broken nptII Gene
  • nptII Gene Detector Construct
  • TEM-1 Broken Gene
  • TEM-1 Gene Detector Construct
  • P. destructans Detector Construct


    • Design

    As seen in figure x, the tdk/kan cassette is flanked by bsaI restriction sites on both its 5’ and 3’ ends. These restriction sites produce sticky ends which can ligate to any DNA sequence possessing the necessary restriction sites with the complementary overhangs. Figure X shows how the tdk/kan cassette can be ligated to DNA sequences homologous to ADP1’s genome.