pAC-LYC
(Plasmid #53270)

• Purpose: Contains 3 genes (crtE, crtI, and crtB) of the carotenoid pathway gene cluster of Erwinia herbicola (Pantoea agglomerans) Eho10 and thereby produces lycopene 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: pAC-EHER
• Backbone manufacturer: Francis X. Cunningham, Jr.
• Backbone size w/o insert (bp): 13581
• Total vector size (bp): 9476
• Modifications to backbone: Constructed from pAC-EHER by deletion of adjacent 0.8 and 1.1 kB BamHI-BamHI frag- ments, as well as and adjacent 1.6 and 0.6 kB AvaI-AvaI fragments. This served to remove most or all of the coding regions of the idi (ORF6), crtX, crtY, and crtZ genes.
• 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. Use to confirm the function of the products of prospective DXS or IPI or LytB-encoding genes or cDNAs. Active enzymes will yield E. coli colonies much darker in color [see Cunningham FX Jr, Lafond TP, Gantt E (2000) Evidence of a role for LytB in the nonmevalonate pathway of isoprenoid biosynthesis. J Bacteriol. 182, 5841-5848.]. Use pAtipiTrc or pdxs/ipiTrc as a positive control.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: crtE, crtI, crtB
• Species: Erwinia herbicola Eho10
• GenBank ID: M87280.1
• Promoter: endogenous promoters

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NA (unknown if destroyed)
• 3′ cloning site: NA (unknown if destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: none

Resource Information

• Articles Citing this Plasmid:
  • 7 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.