PurposeContains Erwinia herbicola Eho10 (Pantoea agglomerans) crtE and crtB genes, and thereby produces phytoene in E. coli
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||53300||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) 13581
- Total vector size (bp) 8106
Modifications to backboneConstructed from pAC-EHER by deletion of adjacent 1.1 kB Sall-Sal1 and 2.0 kB Sall-Bglll fragments, and deletion also of adjacent 0.8 kB and 1.1 kB BamHI-BamHI fragments.
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 phytoene production, or grow on agar plates at room temperature for 3-7 days.
Copy numberLow Copy
Gene/Insert namecrtE, crtB
SpeciesErwinia herbicola Eho10
- Promoter endogenous promoters
- 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)
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-PHYT was a gift from Francis X Cunningham Jr (Addgene plasmid # 53300 ; http://n2t.net/addgene:53300 ; RRID:Addgene_53300)
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