Price Information
Cat No. | Plasmid Name | Availability | Add to cart |
---|---|---|---|
V001191 | 35S:RUBY | In stock (lyophilized plasmid) |
Buy one, get one free! |
Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.
Basic Vector Information
RUBY reporter converts tyrosine to vividly red betalain, which is clearly visible to naked eyes without the need of using special equipment or chemical treatments. Tyrosine is first hydroxylated on the benzene ring, resulting in L-3,4-dihydroxyphenylalanine (L-DOPA). The reaction is catalyzed by the P450 oxygenase CYP76AD1. L-DOPA can be further oxidized into cyclo-DOPA by CYP76AD1. Alternatively, L-DOPA is catalyzed by L-DOPA 4,5-dioxygenase (DODA) into betalamic acid, which is subsequently condensed with cyclo-DOPA into betanidin. The condensation reaction does not require an enzyme. Finally, a sugar moiety is added to betanidin by a glucosyltransferase to generate the colorful betalain. CYP76AD1, DODA, and Glucosyltransferase were organized into a single open reading frame to form the RUBY reporter and the three genes were linked by sequences that encode 2A peptides.
35S:RUBY vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
References
- He, Y., Zhang, T., Sun, H. et al. A reporter for noninvasively monitoring gene expression and plant transformation. Hortic Res 7, 152 (2020). https://doi.org/10.1038/s41438-020-00390-1
35S:RUBY vector Sequence
LOCUS 35S:RUBY. 14335 bp DNA circular SYN 25-DEC-2023 DEFINITION RUBY under the control of CaMV 35S promoter/marker for transformation. ACCESSION . VERSION . KEYWORDS 35S:RUBY. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14335) AUTHORS He Y, Zhang T, Sun H, Zhan H, Zhao Y TITLE A reporter for noninvasively monitoring gene expression and plant transformation. JOURNAL Hortic Res. 2020 Sep 19;7:152. doi: 10.1038/s41438-020-00390-1. eCollection 2020. PUBMED 33024566 REFERENCE 2 (bases 1 to 14335) TITLE Direct Submission REFERENCE 3 (bases 1 to 14335) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Hortic Res."; date: "2020-09-19" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..14335 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 32..708 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" CDS 717..2207 /gene="CYP76AD1" /label=CYP76AD1 /note="Cytochrome P450 76AD1 from Beta vulgaris. Accession#: I3PFJ5" CDS 2217..2273 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 2274..3098 /codon_start=1 /label=4,5-DOPA dioxygenase extradiol alpha 1 /translation="MKMMNGEDANDQMIKESFFITHGNPILTVEDTHPLRPFFETWREK IFSKKPKAILIISGHWETVKPTVNAVHINDTIHDFDDYPAAMYQFKYPAPGEPELARKV EEILKKSGFETAETDQKRGLDHGAWVPLMLMYPEADIPVCQLSVQPHLDGTYHYNLGRA LAPLKNDGVLIIGSGSATHPLDETPHYFDGVAPWAAAFDSWLRKALINGRFEEVNIYES KAPNWKLAHPFPEHFYPLHVVLGAAGEKWKAELIHSSWDHGTLCHGSYKFTSA" CDS 3108..3164 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 3165..4667 /codon_start=1 /label=cyclo-DOPA 5-O-glucosyltransferase /translation="MTAIKMNTNGEGETQHILMIPFMAQGHLRPFLELAMFLYKRSHVI ITLLTTPLNAGFLRHLLHHHSYSSSGIRIVELPFNSTNHGLPPGIENTDKLTLPLVVSL FHSTISLDPHLRDYISRHFSPARPPLCVIHDVFLGWVDQVAKDVGSTGVVFTTGGAYGT SAYVSIWNDLPHQNYSDDQEFPLPGFPENHKFRRSQLHRFLRYADGSDDWSKYFQPQLR QSMKSFGWLCNSVEEIETLGFSILRNYTKLPIWGIGPLIASPVQHSSSDNNSTGAEFVQ WLSLKEPDSVLYISFGSQNTISPTQMMELAAGLESSEKPFLWVIRAPFGFDINEEMRPE WLPEGFEERMKVKKQGKLVYKLGPQLEILNHESIGGFLTHCGWNSILESLREGVPMLGW PLAAEQAYNLKYLEDEMGVAVELARGLEGEISKEKVKRIVEMILERNEGSKGWEMKNRA VEMGKKLKDAVNEEKELKGSSVKAIDDFLDAVMQAKLEPSLQ" terminator 4675..4923 /label=HSP terminator /note="efficient transcription terminator from the Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya et al., 2010)" promoter 5403..5586 /label=NOS promoter /note="nopaline synthase promoter" CDS 5638..6648 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" terminator 6701..6953 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(7528..7545) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7528..7544) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7537..7559) /label=M13/pUC Forward /note="In lacZ gene" misc_feature complement(7766..7790) /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 8311..9099 /label=SmR /note="aminoglycoside adenylyltransferase (Murphy, 1985)" rep_origin 9348..9936 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10090..10107 /label=L4440 /note="L4440 vector, forward primer" misc_feature complement(10122..10262) /label=bom /note="basis of mobility region from pBR322" primer_bind 10348..10370 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin complement(10606..10800) /direction=LEFT /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(10869..11939) /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(12371..12997) /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)"