PurposeContains crtE, crtI, crtY genes of Erwinia herbicola (Pantoea agglomerans) Eho10, and will produce beta-carotene in E. coli when complemented with a gene encoding phytoene synthase
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||53282||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Backbone manufacturerNew England BioLabs
- Backbone size w/o insert (bp) 4245
- Total vector size (bp) 9424
Modifications to backbonePlasmid pAC-BETA was digested with EcoRI and partially digested with BamHI and a 7.1 kB fragment was gel-purified. Plasmid pACYC184 was digested with EcoRI and BamHI and a 2.4 kB fragment was gel-purified. The two fragments were ligated together to give plasmid pAC-85b.
Vector typelow copy number bacterial cloning vector
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol, 25 μg/mL
Growth instructionsUse to confirm the function of the products of genes or cDNAs that are thought to encode a phytoene synthase enzyme, or to screen genomic or cDNA libraries for plasmids containing such genes (crtB) or cDNAs (psy). The plasmid pAdPSY1 can be used as a positive control. Grow in liquid culture on a platform shaker at 28 degrees Celsius in darkness for 2-3 days, or on agar plates at room temperature for 3-7 days. A yellow colony color and an accumulation of beta-carotene is indicative of phytoene synthase activity.
Copy numberLow Copy
Gene/Insert namecrtE, crtY, crtI
SpeciesErwinia herbicola Eho10
Insert Size (bp)7048
- Promoter endogenous promoters
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site BamHI (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-85b was a gift from Francis X Cunningham Jr (Addgene plasmid # 53282 ; http://n2t.net/addgene:53282 ; RRID:Addgene_53282)
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