beauty inducible transposon plasmid psbtet gp  (Addgene inc)

Journal: bioRxiv

Article Title: A scalable transposon mutagenesis system for non-model bacteria

doi: 10.64898/2025.12.22.696024

Figure Lengend Snippet: ( A ) Schematic of the transposon variant library components. LE, left mosaic end; RE, right mosaic end; oriT , origin of transfer; ABR, antibiotic resistance. ( B ) Mean transcription strength and GC content (%) of 16 promoters selected for inclusion in the transposon variant library (green). Transcription strength for 421 promoter sequences was measured across 10 bacterial strains by Yim et al. 2019 . ( C ) Himar1-only library variant performance in E. coli DH10B, with either the catP (CAM) or ( D ) aadA(9) (SPEC) ABR gene. Relative transposition efficiency (enrichment) was measured by Tn-seq after pooled conjugative delivery. ( E ) Correlation between previously published transcriptional strength of promoters and their transposition efficiencies in E. coli when driving the Himar1 transposase. ( F ) Correlation between previously published transcriptional strength of promoters and their transposition efficiencies in E. coli when driving the catP ABR.

Article Snippet: All transposon/ABR promoter part plasmids and transposon plasmid destination vectors used in this study have been made available via Addgene (see Supplementary Data Table 1 and Supplementary Data Table 2 for Addgene IDs).

Techniques: Variant Assay

Journal: bioRxiv

Article Title: A scalable transposon mutagenesis system for non-model bacteria

doi: 10.64898/2025.12.22.696024

Figure Lengend Snippet: ( A ) Bacterial strains ranked by the number of unique genomic insertion events measured by Tn-seq analysis. ‘High confidence’ strains received over 100 insertions per sample, while ‘low confidence’ strains received between 10-100. Each data point represents the total number of unique genomic insertions sites obtained from delivery of a 324-member transposon variant library. Note the data for Cupriavidus necator is a combination of two different strains (see Supplementary Table 2) ( B ) Himar1 transposase promoter efficiencies in strains with at least 100 observed insertion events in multiple samples. Each row represents a single replicate, and rows are clustered by relative transposase promoter efficiencies. Strains are colored by their class: Gammaproteobacteria (red), Alphaproteobacteria (green), Betaproteobacteria (purple), or Verruomicrobia (yellow). ( C ) The replicability of promoter efficiencies as measured by Tn-seq across all microbes screened consistently increases with transformation efficiency for both transposase and ABR promoters. ( D ) Validation of promoter efficiencies. Promoters with high (H), medium (M), and low (L) abundances from the screen results were individually cloned and then tested for their conjugation efficiencies in three strains. Conjugation efficiency is reported as the number of transconjugants per recipient.

Article Snippet: All transposon/ABR promoter part plasmids and transposon plasmid destination vectors used in this study have been made available via Addgene (see Supplementary Data Table 1 and Supplementary Data Table 2 for Addgene IDs).

Techniques: Variant Assay, Transformation Assay, Biomarker Discovery, Clone Assay, Conjugation Assay

Journal: bioRxiv

Article Title: A scalable transposon mutagenesis system for non-model bacteria

doi: 10.64898/2025.12.22.696024

Figure Lengend Snippet: ( A ) Number of genomic insertions observed for different recipient strains undergoing dual-transposon promoter screen, as determined by sequencing. Conjugations were performed either under standard petri dish-based format (‘high input’) or 96-well plate-based format (‘low input’) and transconjugants sequenced after growth in selective media. ( B ) Comparison of Tn5 (orange) or Himar1 (blue) promoter abundance between the Himar1-only or Tn5-only transposase library screens (y-axis) and Himar-Tn5 dual-transposase library screen (x-axis). ( C ) Comparing the efficiencies of promoters associated with Himar1 (y-axis) versus Tn5 (x-axis) transposons for three strains.

Article Snippet: All transposon/ABR promoter part plasmids and transposon plasmid destination vectors used in this study have been made available via Addgene (see Supplementary Data Table 1 and Supplementary Data Table 2 for Addgene IDs).

Techniques: Sequencing, Comparison