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(Plasmid #53288)


Item Catalog # Description Quantity Price (USD)
Plasmid 53288 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.


  • Vector backbone
  • Backbone manufacturer
    Francis X. Cunningham, Jr.
  • Backbone size w/o insert (bp) 9476
  • Total vector size (bp) 11304
  • Modifications to backbone
    The entire open reading frame of an Arabidopsis lcyB cDNA [GenBank U50739.1] in plasmid pZL1 was excised as an EcoRI-HindIII fragment of ca. 1.8 kB and cloned in the same sites of pBluescript SK- so that the cDNA was fused, in frame, to the N terminal portion of the lacZ gene and under the control of the lac promoter. From this plasmid (pAtLCYbSK) a ca. 1.8-kB NotI-NotI fragment containing the lcyB cDNA (without the lac promoter) was excised, made blunt with the Klenow enzyme, and cloned in the Klenow-blunted HindIII site of pAC-LYC. The lcyB cDNA in this 11.3 kB plasmid was ascertained, by restriction enzyme digestion, to be in the forward orientation. Despite the lack of a bacterial promoter, production of the LCYb enzyme is sufficient so that beta-carotene is the major carotenoid in E. coli, but also low enough so that the introduction of a second plasmid with a strongly-expressed lycopene epsilon cyclase cDNA (i.e plasmid pY2F) will yield predominantly alpha-carotene in E. coli.
  • Vector type
    low copy number bacterial cloning vector

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol, 25 μg/mL
  • Growth Temperature
  • Growth Strain(s)
  • 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. Combine with plasmid pY2F and grow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness for best alpha-carotene production, or grow on agar plates at room temperature for 3-7 days.
  • Copy number
    Low Copy


  • Gene/Insert name
  • Alt name
    lycopene beta-cyclase
  • Species
    A. thaliana (mustard weed)
  • Insert Size (bp)
  • GenBank ID
  • Entrez Gene
    LYC (a.k.a. AT3G10230, LYCOPENE BETA-CYCLASE, lycopene cyclase)
  • Promoter none

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (destroyed during cloning)
  • 3′ cloning site HindIII (destroyed during cloning)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
  • Zeocin® is an InvivoGen trademark.

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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pAC-BETA-At was a gift from Francis X Cunningham Jr (Addgene plasmid # 53288 ; ; RRID:Addgene_53288)
  • For your References section:

    Carotenoid biosynthesis in the primitive red alga Cyanidioschyzon merolae. Cunningham FX Jr, Lee H, Gantt E. Eukaryot Cell. 2007 Mar;6(3):533-45. Epub 2006 Nov 3. 10.1128/EC.00265-06 PubMed 17085635