pYTRW26K_1Ti1
(Plasmid #177292)

• Purpose: genomic integration of genes (inserted in I-SceI-site) via rrn-interposon/SacB with TcR and eYFP
• Depositing Lab: Karl-Erich Jaeger
• Publication: Weihmann et al FEMS Microbes. 2022 Dec 19;4:xtac030. doi: 10.1093/femsmc/xtac030. eCollection 2023.

Backbone

• Vector backbone: pYTRW21K_1Ti1
• Backbone size w/o insert (bp): 11167
• Vector type: Bacterial Expression, Synthetic Biology ; Yeast Expression, rrn genomic integration
• Selectable markers: URA3 ; Tetracycline

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: PsacB-sacB
• Alt name: sucrose sensitivity module
• Alt name: Levansucrase
• Species: Bacillus subtilis
• Insert Size (bp): 1884

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: CGATTGACGGCATTACG
• 3′ sequencing primer: GTCTGCGTAGAATCCTC, TTAAAGACGCGGCCAGC

Resource Information

• Supplemental Documents:
  • pYTRW26K_1Ti1.gb
• A portion of this plasmid was derived from a plasmid made by: Schaefer et al (pK19-mobSacB in PMID: 8045426)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Part of the pYT vector series which facilitates chromosomal interposon integration in rrn "landing pads" with selection on tetracyline and transcriptional reporter eYFP (https://www.fpbase.org/protein/eyfp/; methods: flow cytometry, fluorescence visualisation, fluorescence measurements). Target genes can be inserted at the I-SceI site. The vector is equipped with SacB for counter selection.