PurposeContains the Erwinia herbicola (Pantoea agglomerans) Eho10 genes crtE and crtB, and the Rhodobacter capsulatus crtI gene and thereby produces neurosporene in Escherichia coli
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||53271||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) 8106
- Total vector size (bp) 11849
Modifications to backboneA 3.7 kB EcoRV-EcoRV fragment containing the phytoene desaturase gene from Rhodobacter capsulatus (crtI) was excised from plasmid GABX2 (See: Armstrong, G.A., Alberti, M., Leach, F., and Hearst, J.E. (1989). Nucleotide sequence, organization, and nature of the protein products of the carotenoid biosynthesis gene cluster of Rhodobacter capsulatus. Mol. Gen. Genet. 216, 254-268.) and inserted in the Klenow-blunted SalI site of pAC-PHYT.
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 neurosporene production, or grow on agar plates at room temperature for 3-7 days.
Copy numberLow Copy
Alt namephytoene desaturase
- Promoter endogenous promoters
- Cloning method Restriction Enzyme
- 5′ cloning site SalI (destroyed during cloning)
- 3′ cloning site SalI (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-NEUR was a gift from Francis X Cunningham Jr (Addgene plasmid # 53271 ; http://n2t.net/addgene:53271 ; RRID:Addgene_53271)
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