GGDestX2-Amp
(Plasmid
#157654)
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Purpose(Empty Backbone) Golden Gate assembly and mammalian expression of dCas9 fusion-proteins plus gRNA
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Depositing Lab
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Publication
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 157654 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
This material is available to academics and nonprofits only.
Backbone
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Vector backbonepX330
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Backbone manufacturerFeng Zhang lab
- Backbone size (bp) 4267
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Modifications to backbonepX330 was linearized with BbsI. A dsOligo insert (BsaI, BsaI) with complementary 4 bp overhangs was ligated into the pX330 backbone. This construct was linearized with AgeI and SacI to remove the Cas open reading frame and replace it with a BbsI, BbsI dsOligo insert.
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Vector typeMammalian Expression, Synthetic Biology
- Promoter U6, CBh
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
Cloning Information
- Cloning method Gateway Cloning
- 3′ sequencing primer BGHR (5'-tagaaggcacagtcgagg) (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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 annotated sequence is also available at the Haynes lab Benchling site: https://benchling.com/s/seq-a5tlIDUJ212rLoJgH6yR
How to cite this plasmid
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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:
GGDestX2-Amp was a gift from Karmella Haynes (Addgene plasmid # 157654 ; http://n2t.net/addgene:157654 ; RRID:Addgene_157654) -
For your References section:
Rapid Single-Pot Assembly of Modular Chromatin Proteins for Epigenetic Engineering. Priode JH, Haynes KA. Protocols.io, Jan 19, 2021 10.17504/protocols.io.brgcm3sw
Map uploaded by the depositor.
