URL: http://www.addgene.org/66617
Proper Citation: RRID:Addgene_66617
Insert Name: TPST1
Organism: Homo sapiens
Bacterial Resistance: Kanamycin
Defining Citation: PMID:18522538
Vector Backbone Description: Backbone Marker:Clontech; Vector Backbone:pEGFP-N1; Vector Types:Mammalian Expression; Bacterial Resistance:Kanamycin
Comments: Molecular cloning of TPST (tyrosylprotein sulfotransferase)–EGFP (enhanced green fluorescent protein) The TPST1 and TPST2 sequences were obtained from human ESTs (expressed sequence tags) from the RZPD (Deutsches Ressourcenzentrum für Genomforschung, Berlin, Germany; GenBank® accession numbers NM_003596 and AF061254 respectively). TPST1–EGFP and TPST2–EGFP were obtained by PCR using the following primers: plus strand, 5′-GGGGCTCGAGATGGTTGGAAAGCTGAAG-3′, and minus strand, 5′-GGGGCTGCAGCTCCACTTGCTCAGTC-3′ for TPST1; and plus strand, 5′-GGGGCTCGAGATGCGCCTGTCGGTGC-3′, and minus strand, 5′-GGGGCTGCAGCGAGCTTCCTAAGTGG-3′ for TPST2 (XhoI and PstI, restriction sites are underlined). PCR products were purified and ligated into the pGEM-T-Easy vector (Promega) following the manufacturer's instructions. Positive clones were digested with XhoI and PstI, and the fragments obtained were inserted in the corresponding sites of pEGFP-N1 (Clontech). All constructs were verified by double-strand DNA sequencing (MWG Biotech, Ebersburg, Germany). Two differences to the published sequence of TPST2 were identified (276C>T and 1002T>C, relative to the start codon), but neither of which affect the predicted protein sequence.
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Source: Addgene (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
URL: http://www.addgene.org
