pSAM_Cam
(Plasmid #91570)

• Purpose: (Empty Backbone) Mariner transposon mutagenesis & sequencing vector - Transposon contains KanR and Illumina sequencing adapters. Lac promoter expresses transposase. Chloramphenicol resistance on backbone.
• Depositing Lab: Harry Mobley
• Publication: Bachman et al MBio. 2015 Jun 9;6(3):e00775. doi: 10.1128/mBio.00775-15.

Backbone

• Vector backbone: pSAM_Ec
• Backbone manufacturer: N/A
• Backbone size (bp): 4559
• Modifications to backbone: Replaced ampicillin resistance with chloramphenicol
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Kanamycin, 25 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): S17-1λpir
• Growth instructions: Add 10 mM glucose to repress transposase. S17-1λpir may be EndA+ so extra purification steps may be necessary during miniprep.
• Copy number: Low Copy

Resource Information

• Supplemental Documents:
  • pSam_Cam.gbk
  • pSAM_Cam seqbuilder file

Terms and Licenses

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

Depositor Comments

Mariner himar1C9 transposase driven by lac promoter on an R6K gamma suicide plasmid. The transposon allows identification of fitness factors by Illumina sequencing. This transposon will randomly insert a kanamycin resistance cassette into genomic TA sites. Chloramphenicol resistance is contained on the plasmid backbone.

For further background, see:
Goodman, Andrew L., Meng Wu, and Jeffrey I. Gordon. “Identifying Microbial Fitness Determinants by Insertion Sequencing (INSeq) Using Genome-Wide Transposon Mutant Libraries.” Nature Protocols 6.12 (2011): 1969–1980. PMC. Web. 11 Apr. 2017.

Wiles, Travis J. et al. “Combining Quantitative Genetic Footprinting and Trait Enrichment Analysis to Identify Fitness Determinants of a Bacterial Pathogen.” Ed. Danielle A. Garsin. PLoS Genetics 9.8 (2013): e1003716. PMC. Web. 11 Apr. 2017.