PurposeContains ctrE, crtB, crtI, crtY, and idi genes of Erwinia herbicola (Pantoea agglomerans) Eho10 and thereby produces beta-carotene in Escherichia coli
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||53277||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) 10609
- Total vector size (bp) 9259
Modifications to backboneA 1.6 kb SphI-DrdI fragment was deleted from pAC-BETA to give pAC-BETA9.0 (unpublished). A 2.6 kb HindIII-blunted SphI fragment, containing CrtE and idi (ORF6), a gene encoding a type 2 isopentenyl diphosphate isomerase, was excised from pAC-EHER and inserted into the HindIII-Klenow-blunted SalI sites of pAC-BETA9.0 to give pAC-BETAipi.
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 beta-carotene production, or grow on agar plates at room temperature for 3-7 days.
Copy numberLow Copy
Alt nameisopentenyl diphosphate isomerase
SpeciesErwinia herbicola Eho10
- Promoter endogenous promoter
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site SalI (not 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-BETAipi was a gift from Francis X Cunningham Jr (Addgene plasmid # 53277 ; http://n2t.net/addgene:53277 ; RRID:Addgene_53277)
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