pAC-EHER
(Plasmid #53262)

• Purpose: Contains a 7 gene carotenoid pathway gene cluster of Erwinia herbicola (Pantoea agglomerans) Eho10 and thereby produces zeaxanthin diglucoside 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: pACYC184
• Backbone manufacturer: New England BioLabs
• Backbone size w/o insert (bp): 4245
• Total vector size (bp): 13581
• Modifications to backbone: A 9.34 kB BglII-BglII genomic fragment, containing a seven gene carotenoid pathway cluster from Erwinia herbicola (Pantoea agglomerans) Eho10, was excised from plasmid pPL376, and inserted into the BamHI site of pACYC184 in the forward orientation.
• 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 carotenoid production, or grow on agar plates at room temperature for 3-7 days.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: crtE, idi, crtX, crtY, crtI, crtB, crtZ
• Alt name: the idi gene was earlier referred to as ORF6
• Species: Erwinia herbicola Eho10
• Insert Size (bp): 9340
• GenBank ID: M87280.1
• Promoter: endogenous promoters

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The E. herbicola carotenoid gene cluster was excised from plasmid pPL376, obtained from the late Robert W. Tuveson. See: 1. Tuveson, R.W., Larson, R.A., and Kagan, J. (1986) Role of cloned carotenoid genes expressed in Escherichia coli in protecting against inactivation by near-UV light and specific phototoxic molecules. J. Bacteriol. 170, 4675-4680. 2. Perry, K.L., Simonitch, T.A., Harrison-Lavoie, K. J., and Liu, S. (1986) Cloning and regulation of frwinia herbicola pigment genes. J. Bacteriol. 168, 607-612. 3. Hundle,B., Alberti,M., Nievelstein,V., Beyer,P., Kleinig,H., Armstrong,G.A., Burke,D.H. and Hearst,J.E.(1994) Functional assignment of Erwinia herbicola Eho10 carotenoid genes expressed in Escherichia coli Mol. Gen. Genet. 245, 406-416.
• Article Citing this Plasmid:
  • 1 Reference

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.