pTE1062
(Plasmid #172356)

• Purpose: Detection of Streptomyces coelicolor butyrolactones (SCB) by kanamycin bioassay. Streptomyces genomic integrative vector.
• Depositing Lab: Eriko Takano
• Publication: Biarnes-Carrera et al Methods Mol Biol. 2018;1673:117-128. doi: 10.1007/978-1-4939-7309-5_10.

Backbone

• Vector backbone: pSET152
• Backbone size w/o insert (bp): 5715
• Total vector size (bp): 9186
• Modifications to backbone: Cloning of ScbR under native promoter from Streptomyces coelicolor chromosome. Insertion of kanamycin resistance gene under regulation of kasOp promoter (SCB regulated). Insertion of hygromycin resistance gene for selection.
• Vector type: Bacterial Expression, Synthetic Biology ; Integrative into Streptomyces genome via serine integrase (like parent pSET152)

Growth in Bacteria

• Bacterial Resistance(s): Hygromycin, 200 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: scbR
• Alt name: SCO6265
• Insert Size (bp): 1240
• Entrez Gene: SCO6265 (a.k.a. SCO6265, SCAH10.30c, scbR)

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: GCCGAATAGCCTCTCCAC
• 3′ sequencing primer: CTGGCGTAATAGCGAAGAG

Gene/Insert 2

• Gene/Insert name: kasOp
• Alt name: Promoter for kasO (SCB regulated, positioned to express kanR)
• Species: Streptomyces coelicolor
• Insert Size (bp): 100

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: GCCGAATAGCCTCTCCAC
• 3′ sequencing primer: CTGGCGTAATAGCGAAGAG

Resource Information

• Supplemental Documents:
  • pTE1062.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Biarnes-Carrera, Marc, Chang Kwon Lee, Takuya Nihira, Rainer Breitling, and Eriko Takano. ‘Orthogonal Regulatory Circuits for Escherichia Coli Based on the γ-Butyrolactone System of Streptomyces Coelicolor’. ACS Synthetic Biology 7, no. 4 (2018): 1043–55. https://doi.org/10.1021/acssynbio.7b00425.

pTE1062 is replicative in E. coli via pMB1 type origin, and carrying oriT (RP4) for delivery to Streptomyces by conjugation. In Streptomyces, pTE1062 is not replicative, and therefore selection is on integration into the genome mediated by phiC31 integrase.