Part:BBa_K4342000
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
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.| None |