pDY4
(Plasmid #53589)

• Purpose: Produces an active Lactuca sativa (lettuce) lycopene epsilon-cyclase in E. coli. Use with pAC-LYC or pAC-LYCipi to produce epsilon-carotene in E. coli.
• Depositing Lab: Francis X Cunningham Jr
• Publication: Cunningham FX et al Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2905-10. Epub 2001 Feb 20.

Backbone

• Vector backbone: pBluescript SK-
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 2958
• Total vector size (bp): 4762
• Modifications to backbone: This is a cDNA library plasmid containing a Lactuca sativa (romaine lettuce) lycopene epsilon-cyclase cDNA. This plasmid has been referred to elsewhere as pLsLCYe [Cunningham FX Jr, Gantt E (2001) A portfolio of plasmids for identification and analysis of carotenoid pathway enzymes: Adonis aestivalis as a case study. Photosynth Res. 92, 245-259.].
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Growth instructions: Can be used with pAC-LYC or pAC-LYCipi in E. coli strain TOP10 to produce epsilon-carotene. For this purpose, grow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness, or grow on agar plates at room temperature for 3-7 days.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: lcyE
• Alt name: DY4
• Alt name: lycopene epsilon-cyclase
• Alt name: lycopene epsilon-bicyclase
• Species: Lactuca sativa
• Insert Size (bp): 1780
• GenBank ID: AF321538.1
• Promoter: lac

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: NA
• 3′ sequencing primer: NA

Resource Information

• A portion of this plasmid was derived from a plasmid made by: This plasmid was selected in a screen of a Lactuca sativa (Romaine lettuce) cDNA library. The library was kindly provided to us by Harry Yamamoto and Bob Bugos of the University of Hawaii [see Bugos RC, Yamamoto HY (1996) Molecular cloning of violaxanthin de-epoxidase from romaine lettuce and expression in Escherichia coli. Proc Natl Acad Sci USA. 93, 6320-6325.].

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please note that the DY4 insert in this plasmid contains an extra aspartic acid, D178, compared to GenBank reference sequence AF321538.1