PurposeContains ctrE, crtB, and crtI genes of Erwinia herbicola (Pantoea agglomerans) Eho10, and the lcyE gene of Lactuca sativa and thereby produces epsilon-carotene in Escherichia coli
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||53276||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Backbone manufacturerFrancis X. Cunningham, Jr.
- Backbone size w/o insert (bp) 9476
- Total vector size (bp) 11729
Modifications to backboneA 2.28 kb PvuII-PvuII fragment with the Lactuca sativa (romaine lettuce) lycopene ɛ-cyclase was excised from the original cDNA library plasmid pDY4 [Cunningham FX Jr, Gantt E (2001) One ring or two? Determination of ring number in carotenoids by lycopene epsilon-cyclases. Proc Natl Acad Sci USA. 98, 2905-2910] and ligated in the Klenow-blunted SalI site of pAC-LYC. Orientation of the inserted lettuce lcyE cDNA was ascertained by restriction enzyme digestion.
Vector typelow copy number bacterial cloning vector
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol, 25 μg/mL
Growth instructionsGrow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness for best epsilon-carotene production, or grow on agar plates at room temperature for 3-7 days.
Copy numberLow Copy
Alt namelycopene epsilon bicyclase
Insert Size (bp)2249
- Promoter lac
- 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)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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.
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-EPSILON was a gift from Francis X Cunningham Jr (Addgene plasmid # 53276 ; http://n2t.net/addgene:53276 ; RRID:Addgene_53276)
For your References section:A study in scarlet: enzymes of ketocarotenoid biosynthesis in the flowers of Adonis aestivalis. Cunningham FX Jr, Gantt E. Plant J. 2005 Feb;41(3):478-92. 10.1111/j.1365-313X.2004.02309.x PubMed 15659105