pAC-ZEAXipi
(Plasmid #53287)

• Purpose: Contains crtE, idi, crtI, crtY, crtB, and crtZ genes of Erwinia herbicola (Pantoea agglomerans) Eho10 and thereby produces zeaxanthin in E. coli
• Depositing Lab: Francis X Cunningham Jr
• Publication: Cunningham FX et al Photosynth Res. 2007 May;92(2):245-59. Epub 2007 Jul 17.

Backbone

• Vector backbone: pAC-EHER
• Backbone manufacturer: Francis X. Cunningham, Jr.
• Backbone size w/o insert (bp): 13581
• Total vector size (bp): 10040
• Modifications to backbone: pAC-EHER was digested with DrdI and partially digested with BsaI, and a resulting 10.64 kB fragment was gel-purified, blunted with the Klenow enzyme, and ligated to give pAC-EHER10.6 (unpublished). This latter plasmid was then digested with SphI, partially digested with DraIII, the ends were made blunt with the Klenow enzyme, and a gel-purified 10.04 kB fragment was then ligated to give pAC-ZEAXipi.
• 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 in darkness for 2-3 days for best zeaxanthin production, or grow on agar plates at room temperature for 3-7 days.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: crtE, idi, crtY, crtI, crtB, crtZ
• Alt name: idi also known as ORF6
• Species: Erwinia herbicola Eho10
• GenBank ID: M87280.1
• Promoter: endogenous promoters

Cloning Information

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

Resource Information

• Articles Citing this Plasmid:
  • 3 References

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.