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pY2F
(Plasmid #53285)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 53285 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBluescript SK-
  • Backbone manufacturer
    Stratagene
  • Backbone size w/o insert (bp) 2958
  • Total vector size (bp) 4884
  • Modifications to backbone
    A cDNA library plasmid referred to as y2 [Cunningham,F.X. Jr., Pogson,B., Sun,Z., McDonald,K.A., DellaPenna,D. and Gantt,E. (1996) Functional analysis of the beta and epsilon lycopene cyclase enzymes of Arabidopsis reveals a mechanism for control of cyclic carotenoid formation. Plant Cell 8, 1613-1626.] contained an Arabidopsis thaliana lcyE cDNA in the reverse orientation relative to the lac promoter of the cloning vector (pBluescript SK-). To improve expression, the cDNA was excised from plasmid y2 as an XhoI-SmaI fragment of 1.9 kB and ligated in pBluescript SK- that had been digested with XhoI, blunted with the Klenow enzyme and then digested with SalI. The resulting plasmid contained the cDNA in the forward orientation relative to the lac promoter.
  • 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
    When using together with plasmid pAC-BETA or pAC-BETAipi to produce alpha-carotene, grow liquid cultures at 28 degrees Celsius in darkness for 2-3 days , or grow on agar plates at room temperature for 3-7 days.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    lcyE
  • Alt name
    lycopene epsilon-cyclase
  • Species
    A. thaliana (mustard weed)
  • Insert Size (bp)
    1929
  • GenBank ID
    U50738.1
  • Entrez Gene
    LUT2 (a.k.a. AT5G57030, LUTEIN DEFICIENT 2, LYCOPENE EPSILON-CYCLASE)
  • Promoter lac

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SalI (destroyed during cloning)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer M13pUC-rev
  • 3′ sequencing primer M13pUC-fwd
  • (Common Sequencing Primers)

Terms and Licenses

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

    pY2F was a gift from Francis X Cunningham Jr (Addgene plasmid # 53285 ; http://n2t.net/addgene:53285 ; RRID:Addgene_53285)
  • For your References section:

    A portfolio of plasmids for identification and analysis of carotenoid pathway enzymes: Adonis aestivalis as a case study. Cunningham FX Jr, Gantt E. Photosynth Res. 2007 May;92(2):245-59. Epub 2007 Jul 17. 10.1007/s11120-007-9210-0 PubMed 17634749