pMCh-623
(Plasmid
#92266)
-
PurposePlasmid for expression of NHA-tagged Drosophila melanogaster GW182 6W6A with six mutated W-motifs in its N-terminal domain in D. melanogaster cells
-
Depositing Labs
-
Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
---|---|---|---|---|---|
Plasmid | 92266 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
-
Vector backbonepAC5.1-NHA
- Backbone size w/o insert (bp) 5445
- Total vector size (bp) 9619
-
Vector typeInsect Expression
Growth in Bacteria
-
Bacterial Resistance(s)Ampicillin, 100 μg/mL
-
Growth Temperature37°C
-
Growth Strain(s)DH5alpha
-
Copy numberHigh Copy
Gene/Insert
-
Gene/Insert namedGW182 6W6A
-
Alt nameGawky
-
SpeciesD. melanogaster (fly)
-
Insert Size (bp)4164
-
MutationThe following point mutations were introduced W239A; W293A; W316A; W338A; W438A; W484A.
-
GenBank IDNM_166783.2
-
Entrez Genegw (a.k.a. Dmel_CG31992, CG11484, CG31992, CG9905, Dmel\CG31992, GQ182, GW, GW182, Gw, Gw182, anon-EST:Posey205, dGW182, gw182)
- Promoter Ac5
-
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
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Note: Addgene's quality control sequencing finds two additional amino acid residue substitutions (V704A and N799S). The effects of these additional residue changes on protein function are not known.
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.
-
For your Materials & Methods section:
pMCh-623 was a gift from Marina Chekulaeva & Witold Filipowicz (Addgene plasmid # 92266 ; http://n2t.net/addgene:92266 ; RRID:Addgene_92266) -
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