AICSDP-4:TUBA1B-mEGFP | Addgene 87421 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 :

AICSDP-4:TUBA1B-mEGFP

catalog :

87421

citations: 2

Reference
Andreu Carbó M, Fernandes S, Velluz M, Kruse K, Aumeier C. Motor usage imprints microtubule stability along the shaft. Dev Cell. 2022;57:5-18.e8 pubmed publisher
Roberts B, Haupt A, Tucker A, Grancharova T, Arakaki J, Fuqua M, et al. Systematic gene tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization. Mol Biol Cell. 2017;28:2854-2874 pubmed publisher

product information

Catalog Number :

87421

Product Name :

AICSDP-4:TUBA1B-mEGFP

article :

doi	10.1091/mbc.E17-03-0209
id	28192202
pubmed_id	28814507

bacterial resistance :

Ampicillin

cloning :

backbone

pUC19

backbone_mutation

backbone_origin

New England BioLabs

backbone_size

promoter

sequencing_primer_3

sequencing_primer_5

vector_types

Mammalian Expression

CRISPR

Other

Donor Template

growth notes :

This plasmid has been used with locus-specific CRISPR/Cas9 to add a mEGFP tag to the second exon of human TUBA1B in WTC human induced pluripotent stem cells by the Allen Institute for Cell Science. Linker (AA) sequence: GGSGGS. After protein tagging using this donor template plasmid and CRISPR/Cas9 reagents, transfected cells may exhibit varying intensity levels of fluorescence, likely due to editing precision. To obtain cells of uniform intensity levels, see our protocol for fluorescence-assisted cell sorting and subcloning of transfected cells (https://www.allencell.org/instructional-videos-and-tutorials-for-cell-methods.html). Further, we recommend PCR-based assays for identifying precisely edited clones as previously described (https://www.molbiolcell.org/doi/abs/10.1091/mbc.e17-03-0209). For more information on the entire plasmid collection, please see https://www.addgene.org/allen-institute-cell-science/ . Please visit https://www.biorxiv.org/content/early/2017/03/31/123042 for bioRxiv preprint.

growth strain :

Homology arms and linker-mEGFP sequence for N-terminus (second exon) tagging of human TUBA1B

origin :

pi :

alt_names

TUBA1B

cloning

clone_method	Gibson Cloning
cloning_site_3
cloning_site_5
promoter
sequencing_primer_3
sequencing_primer_5
site_3_destroyed
site_5_destroyed

entrez_gene

aliases	K-ALPHA-1
gene	TUBA1B
id	10376

genbank_ids

mutation

homology arms contain point mutations to disrupt crRNA binding sites used

name

TUBA1B Homology Arms with mEGFP-linker

shRNA_sequence

size

2750

species

9606

Homo sapiens