pEG608 | Addgene 40194 product information

This webpage contains legacy information. The product is either no longer available from the supplier or has been delisted at Labome.

product summary

company name :

Addgene

product type :

cDNA

product name :

pEG608

catalog :

40194

citations: 1

Reference
Griffin E, Odde D, Seydoux G. Regulation of the MEX-5 gradient by a spatially segregated kinase/phosphatase cycle. Cell. 2011;146:955-68 pubmed publisher

product information

Catalog Number :

40194

Product Name :

pEG608

article :

doi	10.1016/j.cell.2011.08.012
id	5812
pubmed_id	21925318

bacterial resistance :

Ampicillin

cloning :

backbone

pCG150

backbone_mutation

See comment below

backbone_origin

Addgene plasmid 17247

backbone_size

6351

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Worm Expression

growth notes :

Dendra::MEX-5 constructs were constructed as follows. A 4.4 kb mex-5 promoter fragment based on Tenlen et al. (2008) was cloned into pDONRP4P1R (Invitrogen). Dendra2/TEV/S-peptide (Gallo et al., 2010) was cloned into pDONR201. A MEX-5 genomic fragment from the start ATG through 648 bp of 3'UTR was cloned into pDONRP2RP3. Exon 2 of mex-5 was recoded to be RNAi-resistant (Gen-Script) so as to allow depletion of endogenous MEX-5/6 without depletion of the transgene. These constructs were assembled into pCG150 using three-way Gateway system (LR reaction) (Invitrogen) (Merritt et al., 2008). Mutations were made by recombinant PCR and all inserts were sequenced verified. GFP::MEX-5 transgenes were constructed in the same way as Dendra::MEX-5 constructs except a pDONR201-GFP/TEV/FLAG entry clone was used and wild-type sequence of MEX-5 exon 2 was used. Alternate plasmid name: mex-5 prom::GFP/TEV/FLAG::MEX-5::mex-5 3'UTR

origin :

pi :

alt_names

cloning

clone_method	Gateway Cloning
cloning_site_3
cloning_site_5
promoter	4.4 kb mex-5 promoter fragment
sequencing_primer_3
sequencing_primer_5	GFP-F
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	CELE_W02A2.7
gene	mex-5
id	178296

genbank_ids

NM_070165.4

NP_502566.1

mutation

Exon 2 recoded to be RNAi-resistant

name

Mex-5 (RNAi resistant)

shRNA_sequence

size

species

6239

Caenorhabditis elegans