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Addgene

pAC-ZETA
(Plasmid #53316)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 53316 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pAC-PHYT
  • Backbone manufacturer
    Francis X. Cunningham, Jr.
  • Backbone size w/o insert (bp) 8106
  • Total vector size (bp) 10441
  • Modifications to backbone
    A 2.25 kB Pvull-BamHI fragment containing the Synechococcus sp PCC7942 phytoene desaturase gene, pds (crtP), fused to the N terminal portion of the lacZ gene with promoter, was excised from pPDSdel35 [Linden H, Misawa N, Chamovitz D, Pecker I, Hirschberg J, Sandmann G (1991) Functional complementation in Escherichia coli of different phytoene desaturase genes and analysis of accumulated carotenes. Z Naturforsch C. 46, 1045-1051.], blunted with the Klenow enzyme, and ligated in the Klenow-blunted SalI site of pAC-PHYT. The pAC-ZETAipi plasmid produces a mixture of cis isomers of zeta-carotene in E. coli, predominantly 9,15,9′-tri-cis-zeta-carotene and 9,9′-di-cis-zeta-carotene [see Isaacson T, Ohad I, Beyer P, Hirschberg J (2004) Analysis in vitro of the enzyme CRTISO establishes a poly-cis-carotenoid biosynthesis pathway in plants. Plant Physiol. 136, 4246-4255.]
  • 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 zeta-carotene production, or grow on agar plates at room temperature for 3-7 days.
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    pds
  • Alt name
    crtP
  • Species
    Synechococcus PCC7942
  • Insert Size (bp)
    2249
  • GenBank ID
    X55289.1
  • Promoter lazZ

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SalI (not destroyed)
  • 3′ cloning site SalI (destroyed during cloning)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Plasmid pPDSdel35 [Linden H, Misawa N, Chamovitz D, Pecker I, Hirschberg J, Sandmann G (1991) Functional complementation in Escherichia coli of different phytoene desaturase genes and analysis of accumulated carotenes. Z Naturforsch C. 46, 1045-1051.] was obtained from Dr. Joseph Hirschberg of the Hebrew University of Jerusalem.
  • Article Citing this Plasmid

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • 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-ZETA was a gift from Francis X Cunningham Jr (Addgene plasmid # 53316 ; http://n2t.net/addgene:53316 ; RRID:Addgene_53316)
  • For your References section:

    Molecular structure and enzymatic function of lycopene cyclase from the cyanobacterium Synechococcus sp strain PCC7942. Cunningham FX Jr, Sun Z, Chamovitz D, Hirschberg J, Gantt E. Plant Cell. 1994 Aug;6(8):1107-21. 10.1105/tpc.6.8.1107 PubMed 7919981