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pAC-LYC
(Plasmid #53270)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 53270 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

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
  • 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
  • (Common Sequencing Primers)

Resource Information

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-LYC was a gift from Francis X Cunningham Jr (Addgene plasmid # 53270 ; http://n2t.net/addgene:53270 ; RRID:Addgene_53270)
  • 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