pAC-PHYTipi
(Plasmid #53283)

• Purpose: Contains crtE, crtB, and idi genes of Erwinia herbicola (Pantoea agglomerans) Eho10, and thereby produces phytoene 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): 6103
• Modifications to backbone: Plasmid pAC-EHER was digested with DrdI and then partially digested with DraIII, and a 9.46 kB fragment was gel-purified, blunted with the Klenow enzyme, and ligated to give pAC-BETA-CrtX (unpublished). pAC-BETA-CrtX was digested with SphI and BglII, the ends were blunted with mung bean nuclease, and a 6.1 kB fragment was gel-purified and ligated to give pAC-PHYTipi.
• 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 phytoene 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
• 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: BglII (destroyed during cloning)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• 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.