Basic Vector Information
pUB-DEST 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 ℃.
pUB-DEST vector Sequence
LOCUS 40924_44799 10818 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUB-DEST, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10818)
AUTHORS Grefen C, Donald N, Hashimoto K, Kudla J, Schumacher K, Blatt MR.
TITLE A ubiquitin-10 promoter-based vector set for fluorescent protein
tagging facilitates temporal stability and native protein
distribution in transient and stable expression studies
JOURNAL Plant J. 64 (2), 355-365 (2010)
PUBMED 20735773
REFERENCE 2 (bases 1 to 10818)
AUTHORS Grefen C, Donald N, Hashimoto K, Kudla J, Schumacher K, Blatt MR.
TITLE Direct Submission
JOURNAL Submitted (27-NOV-2012) Plant Sciences Group, University of Glasgow,
University Ave, Glasgow G12 8QQ, United Kingdom
REFERENCE 3 (bases 1 to 10818)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10818)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.";
date: "2010"; volume: "64"; issue: "2"; pages: "355-365"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-NOV-2012) Plant Sciences Group, University of Glasgow,
University Ave, Glasgow G12 8QQ, United Kingdom"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10818
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 1..639
/note="ubiquitin 10 promoter from Arabidopsis thaliana
Columbia-6 ecotype (At4g05050)"
/regulatory_class="promoter"
protein_bind 668..792
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 817..847
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 901..1557
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 1902..2204
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(2248..2372)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
regulatory 2449..2657
/label=T35S terminator
/note="T35S terminator"
/regulatory_class="terminator"
promoter 2794..2973
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 3015..3563
/label=BlpR
/note="phosphinothricin acetyltransferase"
terminator 3586..3838
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature complement(3931..3955)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 4476..5264
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
rep_origin 5514..6102
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(6288..6428)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(6772..6966)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(7035..8099)
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(8536..9162)
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
misc_feature complement(10492..10516)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
protein_bind 10681..10702
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 10717..10747
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10755..10771
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 10779..10795
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
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