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product summary
company name :
Addgene
product type :
cDNA
product name :
pSOD1E133insTT-AcGFP1
catalog :
83441
citations: 1
product information
Catalog Number :
83441
Product Name :
pSOD1E133insTT-AcGFP1
article :
| doi | 10.1016/j.cels.2017.10.010 |
| id | 28192374 |
| pubmed_id | 29128334 |
bacterial resistance :
Kanamycin
cloning :
| backbone | pAcGFP1-N1 | |
| backbone_mutation | ||
| backbone_origin | Clontech | |
| backbone_size | 4700 | |
| promoter | ||
| sequencing_primer_3 | ||
| sequencing_primer_5 | ||
| vector_types |
|
growth notes :
Mutation (first described in PMID: 9065559) was created using the method described in PMID: 17446874. Two outer (F: GCTTCGAATTCCATGGCGAC, AcGFP1_R: GGCCATTCACATCGCCATTC) and two mutation-site primers were used. The mutated PCR product and the backbone vector were digested using EcoRI and SalI, gel purified and ligated. SOD1E133insTT mutation introduces a stop codon that disrupts AcGFP1 tag expression. To avoid this, a second round of PCR was done to clone SOD1E133insTT without the stop codon using the same forward primer, and the reverse primer GCTTCGGTCGACCCATCATTTCCACCTTTGCCCAAG, followed by restriction with EcoRI and SalI and ligation in the backbone.
growth strain :
Expresses SOD1E133insTT in mammalian cells.
origin :
37
pi :
|
plasmid copy :
Elizabeth Fisher( pF147 pSOD1A4VAcGFP1, #26408)
resistance markers :
| 3055 |
tags :
High Copy
company information
Addgene
490 Arsenal Way, Suite 100
Watertown, MA 02472
Watertown, MA 02472
info@addgene.org
https://www.addgene.org617.225.9000
headquarters: USA
questions and comments