PurposeIntegrate plasmid of Streptococcus pneumoniae, for chromosome integration of IPTG-inducible dCas9sp
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||85588||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 8273
- Total vector size (bp) 11911
Modifications to backbonegfp_sf gene was removed; dCas9sp gene driven by IPTG inducible promoter PL was inserted
Vector typeBacterial Expression, CRISPR
Growth in Bacteria
Copy numberHigh Copy
Insert Size (bp)4105
- Promoter PLsp
- Cloning method Restriction Enzyme
- 5′ cloning site XbaI (unknown if destroyed)
- 3′ cloning site SalI (unknown if destroyed)
- 5′ sequencing primer TCTCATAGGCCGGCCTGTTAG
- 3′ sequencing primer gctttgctataaatacgcttcacag (Common Sequencing Primers)
To find information of pJWV100, please refer to "Overkamp W, Beilharz K, Detert Oude Weme R, Solopova A, Karsens H, Kovacs A, Kok J, Kuipers OP, Veening JW (2013) Benchmarking various green fluorescent protein variants in Bacillus subtilis, Streptococcus pneumoniae, and Lactococcus lactis for live cell imaging. Applied and environmental microbiology 79: 6481-6490";
To find information of IPTG-inducible promoter, PL, please refer to "Robin Sorg, PhD thesis, Engineering Approaches to Investigate Pneumococcal Gene Expression Regulation and Antibiotic Resistance Development, 2016, ISBN: 978-90-367-9259-2"
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:pJWV102-PL-dCas9 was a gift from Jan-Willem Veening (Addgene plasmid # 85588 ; http://n2t.net/addgene:85588 ; RRID:Addgene_85588)
For your References section:High-throughput CRISPRi phenotyping identifies new essential genes in Streptococcus pneumoniae. Liu X, Gallay C, Kjos M, Domenech A, Slager J, van Kessel SP, Knoops K, Sorg RA, Zhang JR, Veening JW. Mol Syst Biol. 2017 May 10;13(5):931. doi: 10.15252/msb.20167449. PubMed 28490437