pMCh-646
(Plasmid
#92289)
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PurposePlasmid for expression of NHA-tagged Drosophila melanogaster GW182 CED 7W7A with seven mutated W-motifs in D. melanogaster cells
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Depositing Labs
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 92289 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepAC5.1-NHA
- Backbone size w/o insert (bp) 5455
- Total vector size (bp) 6295
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Vector typeInsect Expression
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 namedGW182 CED 7W7A
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Alt nameGawky
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SpeciesD. melanogaster (fly)
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Insert Size (bp)840
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MutationdGW182 CED: aa 940-1215 with the following point mutations: W942A; W1024A; W1037A; W1051A; W1092A; W1107A; W1114A
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GenBank IDNM_166783.2
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Entrez Genegw (a.k.a. Dmel_CG31992, CG11484, CG31992, CG9905, Dmel\CG31992, GQ182, GW, GW182, Gw, Gw182, anon-EST:Posey205, dGW182, gw182)
- Promoter Ac5
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Tag
/ Fusion Protein
- NHA (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site SbfI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer ACA CAA AGC CGC TCC ATC AG
- 3′ sequencing primer CCT TAG AAG GCA CAG TCG AGG (Common Sequencing Primers)
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:
pMCh-646 was a gift from Marina Chekulaeva & Witold Filipowicz (Addgene plasmid # 92289 ; http://n2t.net/addgene:92289 ; RRID:Addgene_92289) -
For your References section:
miRNA repression involves GW182-mediated recruitment of CCR4-NOT through conserved W-containing motifs. Chekulaeva M, Mathys H, Zipprich JT, Attig J, Colic M, Parker R, Filipowicz W. Nat Struct Mol Biol. 2011 Oct 7;18(11):1218-26. doi: 10.1038/nsmb.2166. 10.1038/nsmb.2166 PubMed 21984184