pBBR_Cas9_sp1
(Plasmid
#154851)
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PurposeExpresses SpCas9 and sgRNA that targets the upp gene in Rhodobacter sphaeroides genome
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Depositing Lab
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Depositing OrganizationWageningen University, Department Agrotechnology & Food Sciences
Located in the Netherlands -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 154851 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 154851-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 154851-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepBBRMCS2
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Vector typeCRISPR
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructionsThe plasmid should be transformed to E. coli S17-1, the vector donor strain for R. sphaeroides in diparental conjugation.
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Copy numberUnknown
Gene/Insert 1
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Gene/Insert nameSpcas9 gene
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SpeciesStreptococcus pyogenes
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Insert Size (bp)4122
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MutationCodon harmonized variant for expression in Rhodobacter sphaeroides
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Tag
/ Fusion Protein
- 6xHis (C terminal on insert)
Gene/Insert 2
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Gene/Insert namesgRNA scaffold
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SpeciesStreptococcus pyogenes
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Insert Size (bp)79
Resource Information
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.
Depositor Comments
The plasmid should be transformed to E. coli S17-1, the vector donor strain for R. sphaeroides in diparental conjugation.
DNA (Catalog # 154851-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 154851-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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:
pBBR_Cas9_sp1 was a gift from John van der Oost (Addgene plasmid # 154851 ; http://n2t.net/addgene:154851 ; RRID:Addgene_154851) -
For your References section:
Efficient Cas9-based genome editing of Rhodobacter sphaeroides for metabolic engineering. Mougiakos I, Orsi E, Ghiffary MR, Post W, de Maria A, Adiego-Perez B, Kengen SWM, Weusthuis RA, van der Oost J. Microb Cell Fact. 2019 Nov 25;18(1):204. doi: 10.1186/s12934-019-1255-1. 10.1186/s12934-019-1255-1 PubMed 31767004
