Purpose(Empty Backbone) Mariner transposon mutagenesis & sequencing vector - Transposon contains KanR and Illumina sequencing adapters. Lac promoter expresses transposase. Chloramphenicol resistance on backbone.
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||91570||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 4559
Modifications to backboneReplaced ampicillin resistance with chloramphenicol
Vector typeBacterial Expression
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Kanamycin
Growth instructionsAdd 10 mM glucose to repress transposase. S17-1λpir may be EndA+ so extra purification steps may be necessary during miniprep.
Copy numberLow Copy
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pSAM_Cam was a gift from Harry Mobley (Addgene plasmid # 91570 ; http://n2t.net/addgene:91570 ; RRID:Addgene_91570)
For your References section:Genome-Wide Identification of Klebsiella pneumoniae Fitness Genes during Lung Infection. Bachman MA, Breen P, Deornellas V, Mu Q, Zhao L, Wu W, Cavalcoli JD, Mobley HL. MBio. 2015 Jun 9;6(3):e00775. doi: 10.1128/mBio.00775-15. 10.1128/mBio.00775-15 PubMed 26060277