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product summary
company name :
Addgene
product type :
cDNA
product name :
pAC-VIOL
catalog :
53087
citations: 1
Reference
Neuman H, Galpaz N, Cunningham F, Zamir D, Hirschberg J. The tomato mutation nxd1 reveals a gene necessary for neoxanthin biosynthesis and demonstrates that violaxanthin is a sufficient precursor for abscisic acid biosynthesis. Plant J. 2014;78:80-93 pubmed publisher
product information
Catalog Number :
53087
Product Name :
pAC-VIOL
article :
doi10.1111/tpj.12451
id8242
pubmed_id24506237
bacterial resistance :
Chloramphenicol
cloning :
backbonepAC-ZEAXipi
backbone_mutationAn Arabidopsis thaliana cDNA encoding zeaxanthin epoxidase (zep) was modified so as to delete codons for the first 60 amino acids (the putative chloroplast transit sequence) and cloned in the NdeI and XhoI sites of pET30C so that expression was under the control of the T7 promoter. This plasmid, pET-AtZEP (unpublished), was a gift of Dr. Martin Sergeant, at that time at the University of Warwick. The zep gene with T7 promoter were excised from pET-AtZEP as a Klenow-blunted BspHI-BspHI fragment of ca. 2.4 kB and cloned in the Klenow-blunted AseI site of pAC-ZEAXipi. Insert orientation and sequences at the junctions of the insert were confirmed by DNA sequencing.
backbone_originFrancis X. Cunningham, Jr.
backbone_size10040
promoter
sequencing_primer_3
sequencing_primer_5
vector_types
Other
low copy number bacterial cloning vector
growth notes :
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.
growth strain :
Produces the carotenoid violaxanthin in E. coli.
growth temp :
Grow liquid cultures at 28 degrees Celsius for 2-3 days for best violaxanthin production, or grow on agar plates at room temperature for 3-7 days.
origin :
30
pi :
alt_names
zeaxanthin epoxidase
cloning
clone_methodRestriction Enzyme
cloning_site_3AseI
cloning_site_5AseI
promoterT7
sequencing_primer_3gatggatcttcgaagcgaac
sequencing_primer_5tagccgctaaagcaaacacc
site_3_destroyed1
site_5_destroyed1
entrez_gene
aliasesAT5G67030, ABA DEFICIENT 1, ARABIDOPSIS THALIANA ABA DEFICIENT 1, ARABIDOPSIS THALIANA ZEAXANTHIN EPOXIDASE, ATABA1, ATZEP, IBS3, IMPAIRED IN BABA-INDUCED STERILITY 3, K8A10.10, K8A10_10, LOS6, LOW EXPRESSION OF OSMOTIC STRESS-RESPONSIVE GENES 6, NON-PHOTOCHEMICAL QUENCHING 2, NPQ2, ZEAXANTHIN EPOXIDASE, ZEP
geneABA1
id836838
genbank_ids
AF281655.1
mutationLacks codons for the first 60 N-terminal amino acids that are thought to comprise the chloroplast transit sequence
namezep
shRNA_sequence
size2774
species
3702
Arabidopsis thaliana
tags
plasmid copy :
Plasmid pET-AtZEP, containing a truncated Arabidopsis thaliana zep cDNA under the control of the T7 promoter, was a gift of Dr. Martin Sergeant, at that time of the University of Warwick.
resistance markers :
1973
tags :
Low Copy
company information
Addgene
490 Arsenal Way, Suite 100
Watertown, MA 02472
info@addgene.org
https://www.addgene.org
617.225.9000
headquarters: USA