pAC-NEUR
(Plasmid #53271)

• Purpose: Contains the Erwinia herbicola (Pantoea agglomerans) Eho10 genes crtE and crtB, and the Rhodobacter capsulatus crtI gene and thereby produces neurosporene in Escherichia coli
• Depositing Lab: Francis X Cunningham Jr
• Publication: Cunningham FX et al Plant Cell. 1994 Aug;6(8):1107-21.

Backbone

• Vector backbone: pAC-PHYT
• Backbone manufacturer: Francis X. Cunningham, Jr.
• Backbone size w/o insert (bp): 8106
• Total vector size (bp): 11849
• Modifications to backbone: A 3.7 kB EcoRV-EcoRV fragment containing the phytoene desaturase gene from Rhodobacter capsulatus (crtI) was excised from plasmid GABX2 (See: Armstrong, G.A., Alberti, M., Leach, F., and Hearst, J.E. (1989). Nucleotide sequence, organization, and nature of the protein products of the carotenoid biosynthesis gene cluster of Rhodobacter capsulatus. Mol. Gen. Genet. 216, 254-268.) and inserted in the Klenow-blunted SalI site of pAC-PHYT.
• Vector type: low copy number bacterial cloning vector

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Top10
• Growth instructions: Grow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness for best neurosporene production, or grow on agar plates at room temperature for 3-7 days.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: crtI
• Alt name: phytoene desaturase
• Species: Rhodobacter capsulatus
• GenBank ID: Z11165.1
• Promoter: endogenous promoters

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (destroyed during cloning)
• 3′ cloning site: SalI (destroyed during cloning)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Terms and Licenses

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

Depositor Comments

For better yield of low copy number pAC-based plasmids, grow liquid cultures on a platform shaker at ca. 30 degrees Celsius. When cultures reach early stationary phase, dilute 2-fold with growth medium, add spectinomycin (150 mg/liter), and "amplify" for several hours before harvest. For plasmid selection and maintenance in E. coli, use chloramphenicol at 30 mg/liter.