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URL: http://www.addgene.org/255796
Proper Citation: RRID:Addgene_255796
Insert Name: NrdJ-1C - XtpS
Organism: Xenorhabdus nematophila ATCC 19061
Bacterial Resistance: Spectinomycin and Streptomycin
Vector Backbone Description: Vector Backbone:pCDF; Vector Types:Bacterial Expression; Bacterial Resistance:Spectinomycin and Streptomycin
Comments: This plasmid encodes the termination unit of the non-ribosomal peptide synthetase (NRPS) called xenotetrapeptide synthetase (XtpS), split at an XUTII site and fused to the C-terminal part of the split intein NrdJ-1.When co-expressed with pIN092 and pIN201, the three fragments splice into a functional NRPS that produces the non-ribosomal peptide (NRP) called xenotetrapeptide (XTP; cyclo[Leu-D-Val-Val-D-Val]; CC(C)C[C@H]1C(=O)N[C@@H](C(=O)N[C@H](C(=O)N[C@@H](C(=O)N1)C(C)C)C(C)C)C(C)C; 410.6 g/mol). For successful NRP production, the three plasmids must be co-transformed into an E. coli strain that expresses a broad-spectrum phosphopantetheinyl transferase, such as MtaA (DH10B::mtaA) or Sfp (BAP1), which is required for the post-translational activation of the T domains. Expression is controlled by an araBAD promoter, inducible with 0.2% arabinose.
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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.
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