pAC-LYCipi
(Plasmid #53279)

• Purpose: Contains crtE, crtB, crtI, and idi genes of Erwinia herbicola (Pantoea agglomerans) Eho10, and thereby produces lycopene in Escherichia coli
• Depositing Lab: Francis X Cunningham Jr
• Publication: Cunningham FX et al Eukaryot Cell. 2007 Mar;6(3):533-45. Epub 2006 Nov 3.

Backbone

• Vector backbone: pAC-LYC
• Backbone manufacturer: Francis X. Cunningham, Jr.
• Backbone size w/o insert (bp): 9476
• Total vector size (bp): 8916
• Modifications to backbone: Extraneous sequence was removed from pAC-LYC by digestion with SphI and DrdI, blunting with mung bean nuclease, and recircularization with ligation. This 7.8-kb plasmid was then digested with HindIII and EcoRV to yield a 6.4-kB fragment that was ligated with a HindIII + mung bean-blunted DraIII fragment of 2.55 kB obtained from pAC-EHER.
• 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 lycopene 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: ORF6
• Species: Erwinia herbicola Eho10
• Insert Size (bp): 2750
• GenBank ID: M87280.1
• Promoter: endogenous promoters

Cloning Information

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

Resource Information

• Articles Citing this Plasmid:
  • 2 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.