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UBQ:RUBY vector (V000621)

Price Information

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V000621 UBQ:RUBY In stock, instant shipping

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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 Vector Map

UBQ:RUBY15624 bp70014002100280035004200490056006300700077008400910098001050011200119001260013300140001470015400RB T-DNA repeatP2AP2ANOS promoterNOS terminatorM13 ForwardLB T-DNA repeatSmRoriL4440bompGEX 3'pVS1 oriVpVS1 RepApVS1 StaA

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               15624 bp DNA     circular SYN 12-JUL-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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Hortic 
            Res."; date: "2020-09-19"; volume: "7:152. doi"; pages: " 
            10.1038/s41438-020-00390-1. eCollection 2020"
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"
     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             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"
     promoter        6800..6979
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     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(8957..8974)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8957..8973)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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"