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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
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