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PurposeBacillus subtilis sgRNA expression vector; integrates into amyE
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Depositing Labs
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 79874 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepDG1662
- Backbone size w/o insert (bp) 6982
- Total vector size (bp) 7572
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Modifications to backboneThe rfp-targeting sgRNA RR1 was PCR-amplified from pgRNA-bacteria (Addgene #44251) using primers containing the veg promoter and EcoRI sites, digested with EcoRI, then ligated into either pDG1662 digested with EcoRI to generate pJMP2.
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Vector typeBacterial Expression, CRISPR
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Selectable markersBacillus subtilis chloramphenicol marker
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namesgRNA RR1
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Alt namesgRNA NT
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Alt nameanti-RFP sgRNA
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gRNA/shRNA sequenceAACTTTCAGTTTAGCGGTCT
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SpeciesStreptococcus pyogenes
- Promoter veg
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer agtgattatgccgcgatttc (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byThe rfp-targeting sgRNA RR1 was PCR-amplified from pgRNA-bacteria (Addgene #44251)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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.
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For your Materials & Methods section:
pJMP2 was a gift from Carol Gross & Jason Peters & Stanley Qi (Addgene plasmid # 79874 ; http://n2t.net/addgene:79874 ; RRID:Addgene_79874) -
For your References section:
A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria. Peters JM, Colavin A, Shi H, Czarny TL, Larson MH, Wong S, Hawkins JS, Lu CH, Koo BM, Marta E, Shiver AL, Whitehead EH, Weissman JS, Brown ED, Qi LS, Huang KC, Gross CA. Cell. 2016 Jun 2;165(6):1493-506. doi: 10.1016/j.cell.2016.05.003. Epub 2016 May 26. 10.1016/j.cell.2016.05.003 PubMed 27238023