(364) SB KRAB-SadCas9
(Plasmid
#163022)
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PurposeSleeping Beauty TET-On expression of CRISPRi (without U6-gRNA)
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Depositing Lab
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Depositing OrganizationVanderbilt University
Located in the United States of America -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 163022 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 163022-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 163022-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backboneSleeping Beauty TET-On
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Backbone manufacturerEric Kowarz (Addgene plasmid # 60509)
- Backbone size w/o insert (bp) 5700
- Total vector size (bp) 9206
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Vector typeMammalian Expression ; Transposon
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Selectable markersNeomycin (select with G418)
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)NEB Stable
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Growth instructions34-35 degrees is ideal
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameMammalian codon-optimized SadCas9
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SpeciesS. aureus
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Insert Size (bp)3483
- Promoter TET-On
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Tags
/ Fusion Proteins
- KRAB (N terminal on insert)
- myc NLS (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoR1 (not destroyed)
- 3′ cloning site Not1 (not destroyed)
- 5′ sequencing primer SB-Tre-S
- 3′ sequencing primer SVpa-Seq-A
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byKeith Joung (S. aureus dCas9 from Addgene #70703).
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.
DNA (Catalog # 163022-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 # 163022-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:
(364) SB KRAB-SadCas9 was a gift from Tonia Rex (Addgene plasmid # 163022 ; http://n2t.net/addgene:163022 ; RRID:Addgene_163022) -
For your References section:
Optimization of S. aureus dCas9 and CRISPRi Elements for a Single Adeno-Associated Virus that Targets an Endogenous Gene. Backstrom JR, Sheng J, Wang MC, Bernardo-Colon A, Rex TS. Mol Ther Methods Clin Dev. 2020 Sep 6;19:139-148. doi: 10.1016/j.omtm.2020.09.001. eCollection 2020 Dec 11. 10.1016/j.omtm.2020.09.001 PubMed 33024795
