Basic Vector Information
- Vector Name:
- pUB-Cas9-@GL1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 14834 bp
- Type:
- Binary vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP.
- Promoter:
- CaMV 35S (enhanced)
pUB-Cas9-@GL1 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-@GL1 vector Sequence
LOCUS 40924_44789 14834 bp DNA circular SYN 18-DEC-2018 DEFINITION Binary vector pUB-Cas9-@GL1, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14834) 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 14834) 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 14834) TITLE Direct Submission REFERENCE 4 (bases 1 to 14834) 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..14834 /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..7355 /label=GL1 protospacer sequence /note="GL1 protospacer sequence" misc_RNA 7356..7431 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" regulatory 7556..8195 /label=Arabidopsis thaliana Ubiquitin10 promoter /note="Arabidopsis thaliana Ubiquitin10 promoter" /regulatory_class="promoter" protein_bind 8235..8259 /label=attB1 /note="recombination site for the Gateway(R) BP reaction" CDS 8280..8300 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" CDS 8298..12401 /label=Cas9 /note="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" protein_bind complement(12406..12430) /label=attB2 /note="recombination site for the Gateway(R) BP reaction" terminator 12496..12743 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(12759..12775) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 12783..12799 /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(12807..12837) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(12852..12873) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 13064..13741 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" CDS 13809..14831 /label=HygR /note="aminoglycoside phosphotransferase from E. coli"
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