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PurposeContains 3 genes (crtE, crtI, and crtB) of the carotenoid pathway gene cluster of Erwinia herbicola (Pantoea agglomerans) Eho10 and thereby produces lycopene in Escherichia coli
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Depositing Lab
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Depositing OrganizationUniversity of Maryland, College Park
Located in the United States of America -
Publication
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Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 53270 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 53270-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 53270-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepAC-EHER
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Backbone manufacturerFrancis X. Cunningham, Jr.
- Backbone size w/o insert (bp) 13581
- Total vector size (bp) 9476
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Modifications to backboneConstructed from pAC-EHER by deletion of adjacent 0.8 and 1.1 kB BamHI-BamHI frag- ments, as well as and adjacent 1.6 and 0.6 kB AvaI-AvaI fragments. This served to remove most or all of the coding regions of the idi (ORF6), crtX, crtY, and crtZ genes.
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Vector typelow copy number bacterial cloning vector
Growth in Bacteria
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Bacterial Resistance(s)Chloramphenicol, 25 μg/mL
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Growth Temperature30°C
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Growth Strain(s)Top10
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Growth instructionsGrow liquid cultures on a platform shaker at 28 degrees Celsius for 2-3 days in darkness for best lycopene production, or grow on agar plates at room temperature for 3-7 days. Use to confirm the function of the products of prospective DXS or IPI or LytB-encoding genes or cDNAs. Active enzymes will yield E. coli colonies much darker in color [see Cunningham FX Jr, Lafond TP, Gantt E (2000) Evidence of a role for LytB in the nonmevalonate pathway of isoprenoid biosynthesis. J Bacteriol. 182, 5841-5848.]. Use pAtipiTrc or pdxs/ipiTrc as a positive control.
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Copy numberUnknown
Gene/Insert
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Gene/Insert namecrtE, crtI, crtB
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SpeciesErwinia herbicola Eho10
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GenBank IDM87280.1
- Promoter endogenous promoters
Cloning Information
- 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
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
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.
DNA (Catalog # 53270-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 53270-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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.
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For your Materials & Methods section:
pAC-LYC was a gift from Francis X Cunningham Jr (Addgene plasmid # 53270 ; http://n2t.net/addgene:53270 ; RRID:Addgene_53270) -
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