Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000621 | UBQ:RUBY | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
UBQ:RUBY contains a RUBY under the control of Ubiquitin promoter/marker for transformation.
- Vector Name:
- UBQ:RUBY
- Antibiotic Resistance:
- Spectinomycin, 50 μg/mL
- Length:
- 15624 bp
- Type:
- Plant Expression
- Replication origin:
- ori
- Host:
- Plants
- Promoter:
- NOS
- Cloning Method:
- Gibson Cloning
- 5' Primer:
- CCTGTCAAACACTGATAGTTTctgcagtgcagcgtgacccggtc
- 3' Primer:
- GCTTACTCAGTTAGGTCTAGCTTATCTTTAATCATATTCCATAGTCCA
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
UBQ:RUBY vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- He, Yubing et al. “A reporter for noninvasively monitoring gene expression and plant transformation.” Horticulture research vol. 7,1 152. 19 Sep. 2020, doi:10.1038/s41438-020-00390-1
UBQ:RUBY vector Sequence
LOCUS Exported 15624 bp DNA circular SYN 09-DEC-2024
DEFINITION RUBY under the control of Ubiquitin promoter/marker for
transformation.
ACCESSION .
VERSION .
KEYWORDS UBQ:RUBY
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15624)
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 15624)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 15624)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Hortic
Res."; date: "2020-09-19"; pages: "
10.1038/s41438-020-00390-1. eCollection 2020"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..15624
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 133..157
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
promoter 178..2137
/label=Ubi promoter
/note="Ubiquitin promoter"
CDS 2146..3636
/codon_start=1
/gene="CYP76AD1"
/label=Cytochrome P450 76AD1
/note="Cytochrome P450 76AD1 from Beta vulgaris.
Accession#: I3PFJ5"
/translation="MDHATLAMILAIWFISFHFIKLLFSQQTTKLLPPGPKPLPIIGNI
LEVGKKPHRSFANLAKIHGPLISLRLGSVTTIVVSSADVAKEMFLKKDHPLSNRTIPNS
VTAGDHHKLTMSWLPVSPKWRNFRKITAVHLLSPQRLDACQTFRHAKVQQLYEYVQECA
QKGQAVDIGKAAFTTSLNLLSKLFFSVELAHHKSHTSQEFKELIWNIMEDIGKPNYADY
FPILGCVDPSGIRRRLACSFDKLIAVFQGIICERLAPDSSTTTTTTTDDVLDVLLQLFK
QNELTMGEINHLLVDIFDAGTDTTSSTFEWVMTELIRNPEMMEKAQEEIKQVLGKDKQI
QESDIINLPYLQAIIKETLRLHPPTVFLLPRKADTDVELYGYIVPKDAQILVNLWAIGR
DPNAWQNADIFSPERFIGCEIDVKGRDFGLLPFGAGRRICPGMNLAIRMLTLMLATLLQ
FFNWKLEGDISPKDLDMDEKFGIALQKTKPLKLIPIPRY"
CDS 3646..3702
/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 3703..4527
/codon_start=1
/label=4,5-DOPA dioxygenase extradiol alpha 1
/translation="MKMMNGEDANDQMIKESFFITHGNPILTVEDTHPLRPFFETWREK
IFSKKPKAILIISGHWETVKPTVNAVHINDTIHDFDDYPAAMYQFKYPAPGEPELARKV
EEILKKSGFETAETDQKRGLDHGAWVPLMLMYPEADIPVCQLSVQPHLDGTYHYNLGRA
LAPLKNDGVLIIGSGSATHPLDETPHYFDGVAPWAAAFDSWLRKALINGRFEEVNIYES
KAPNWKLAHPFPEHFYPLHVVLGAAGEKWKAELIHSSWDHGTLCHGSYKFTSA"
CDS 4537..4593
/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 4594..6096
/codon_start=1
/label=cyclo-DOPA 5-O-glucosyltransferase
/translation="MTAIKMNTNGEGETQHILMIPFMAQGHLRPFLELAMFLYKRSHVI
ITLLTTPLNAGFLRHLLHHHSYSSSGIRIVELPFNSTNHGLPPGIENTDKLTLPLVVSL
FHSTISLDPHLRDYISRHFSPARPPLCVIHDVFLGWVDQVAKDVGSTGVVFTTGGAYGT
SAYVSIWNDLPHQNYSDDQEFPLPGFPENHKFRRSQLHRFLRYADGSDDWSKYFQPQLR
QSMKSFGWLCNSVEEIETLGFSILRNYTKLPIWGIGPLIASPVQHSSSDNNSTGAEFVQ
WLSLKEPDSVLYISFGSQNTISPTQMMELAAGLESSEKPFLWVIRAPFGFDINEEMRPE
WLPEGFEERMKVKKQGKLVYKLGPQLEILNHESIGGFLTHCGWNSILESLREGVPMLGW
PLAAEQAYNLKYLEDEMGVAVELARGLEGEISKEKVKRIVEMILERNEGSKGWEMKNRA
VEMGKKLKDAVNEEKELKGSSVKAIDDFLDAVMQAKLEPSLQ"
promoter 6832..7015
/function="NOS promoter"
/label=NOS promoter
/note="nopaline synthase promoter"
terminator 8130..8382
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(8966..8988)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 9195..9219
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 9740..10531
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 10777..11365
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11266..11285
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 11519..11536
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 11551..11691
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(11617..11636)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 11777..11799
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 12035..12229
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(12295..13368)
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
CDS complement(13797..14426)
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"