pAC-BETAipi
(Plasmid #53277)

• Purpose: Contains ctrE, crtB, crtI, crtY, and idi genes of Erwinia herbicola (Pantoea agglomerans) Eho10 and thereby produces beta-carotene in Escherichia coli
• Depositing Lab: Francis X Cunningham Jr
• Publication: Cunningham FX et al Plant J. 2005 Feb;41(3):478-92.

Backbone

• Vector backbone: pAC-BETA
• Backbone manufacturer: Francis X. Cunningham, Jr.
• Backbone size w/o insert (bp): 10609
• Total vector size (bp): 9259
• Modifications to backbone: A 1.6 kb SphI-DrdI fragment was deleted from pAC-BETA to give pAC-BETA9.0 (unpublished). A 2.6 kb HindIII-blunted SphI fragment, containing CrtE and idi (ORF6), a gene encoding a type 2 isopentenyl diphosphate isomerase, was excised from pAC-EHER and inserted into the HindIII-Klenow-blunted SalI sites of pAC-BETA9.0 to give pAC-BETAipi.
• 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 beta-carotene production, or grow on agar plates at room temperature for 3-7 days.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: idi
• Alt name: isopentenyl diphosphate isomerase
• Alt name: ORF6
• Species: Erwinia herbicola Eho10
• GenBank ID: M87280.1
• Promoter: endogenous promoter

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• Articles Citing this Plasmid:
  • 4 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.