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URL: http://www.addgene.org/47498
Proper Citation: RRID:Addgene_47498
Insert Name: cyclin E1
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Defining Citation: PMID:23873023
Vector Backbone Description: Backbone Size:6521; Vector Backbone:MIGR1; Vector Types:Mammalian Expression, Retroviral, MSCV-IRES-GFP; Bacterial Resistance:Ampicillin
Comments: Human cyclin E1 (CCNE1) cDNA was mutated at N- and C-terminal Cdc4 phosphodegrons (T62A and T380A, see references) to disrupt interaction with Fbw7 ubiquitin ligase. cDNA is inserted into MIGR1 (MSCV-based retroviral vector), permitting transduction of hematopoietic cells. Cloned cDNA contains approximately 100 bp from CCNE1 3' UTR. GFP is co-expressed with cDNA via IRES, permitting enrichment for transduced cells using flow sorting. References: Koepp DM, Schaefer LK, Ye X, Keyomarsi K, Chu C, Harper JW et al (2001). Phosphorylation-dependent ubiquitination of cyclin E by the SCFFbw7 ubiquitin ligase. Science 294: 173-177. Strohmaier H, Spruck CH, Kaiser P, Won KA, Sangfelt O, Reed SI (2001). Human F-box protein hCdc4 targets cyclin E for proteolysis and is mutated in a breast cancer cell line. Nature 413: 316-322.
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Source: Addgene
