Basic Vector Information
pUB-Cas9-@BAR 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-Cas9-@BAR vector Sequence
LOCUS 40924_44784 14835 bp DNA circular SYN 18-DEC-2018
DEFINITION Binary vector pUB-Cas9-@BAR, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14835)
AUTHORS Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP.
TITLE An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis
thaliana
JOURNAL Front Plant Sci 8, 39 (2017)
PUBMED 28174584
REFERENCE 2 (bases 1 to 14835)
AUTHORS Hahn F, Mantegazza O, Eisenhut M, Greiner A, Hegemann P, Weber APM.
TITLE Direct Submission
JOURNAL Submitted (03-NOV-2016) Plant Biochemistry, Heinrich Heine
University Dusseldorf, Universitatsstr. 1, Dusseldorf 40225, Germany
REFERENCE 3 (bases 1 to 14835)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 14835)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Front Plant
Sci 8, 39 (2017)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(03-NOV-2016) Plant Biochemistry, Heinrich Heine University
Dusseldorf, Universitatsstr. 1, Dusseldorf 40225, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14835
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 40..214
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature complement(292..316)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 741..1532
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 1622..2210
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2396..2536)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(2880..3074)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(3143..4207)
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(4644..5270)
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
misc_feature complement(6570..6594)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 6797..6813
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 6846..6855
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 6850..6859
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 6903..7326
/label=AtU6-26
/note="Arabidopsis U6-26 gene promoter"
misc_feature 7336..7356
/label=bar1 protospacer sequence
/note="bar1 protospacer sequence"
misc_RNA 7357..7432
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 7557..8196
/label=Arabidopsis thaliana Ubiquitin10 promoter
/note="Arabidopsis thaliana Ubiquitin10 promoter"
/regulatory_class="promoter"
protein_bind 8236..8260
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 8281..8301
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 8299..12402
/label=Cas9
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
protein_bind complement(12407..12431)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
terminator 12497..12744
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(12760..12776)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 12784..12800
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
promoter complement(12808..12838)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(12853..12874)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 13065..13742
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 13810..14832
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
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