Basic Vector Information
- Vector Name:
- pLX-B2-XT1-XT2-hCas9
- Antibiotic Resistance:
- Kanamycin
- Length:
- 10749 bp
- Type:
- Cloning vector
- Replication origin:
- pBBR1 oriV
- Host:
- Plants
- Source/Author:
- Pasin F, Bedoya LC, Bernabe-Orts JM, Gallo A, Simon-Mateo C, Orzaez D, Garcia JA.
- Promoter:
- CaMV 35S
pLX-B2-XT1-XT2-hCas9 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 ℃.
pLX-B2-XT1-XT2-hCas9 vector Sequence
LOCUS 40924_29336 10749 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pLX-B2-XT1-XT2-hCas9, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10749)
AUTHORS Pasin F, Bedoya LC, Bernabe-Orts JM, Gallo A, Simon-Mateo C, Orzaez
D, Garcia JA.
TITLE Multiple T-DNA Delivery to Plants Using Novel Mini Binary Vectors
with Compatible Replication Origins
JOURNAL ACS Synth Biol (2017) In press
PUBMED 28657330
REFERENCE 2 (bases 1 to 10749)
AUTHORS Pasin F, Garcia JA.
TITLE Direct Submission
JOURNAL Submitted (26-MAR-2017) Department of Plant Molecular Genetics,
Centro Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid,
Madrid 28049, Spain
REFERENCE 3 (bases 1 to 10749)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10749)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2017) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-MAR-2017) Department of Plant Molecular Genetics, Centro
Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid, Madrid
28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10749
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..24
/label=assembly linker
/note="assembly linker"
regulatory 25..72
/regulatory_class="terminator"
misc_feature 133..157
/label=LB T-DNA repeat
/note="left border repeat from octopine Ach5 T-DNA"
misc_feature 184..208
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
terminator complement(287..539)
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(582..1373)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
promoter complement(1407..1586)
/label=NOS promoter
/note="nopaline synthase promoter"
regulatory 1677..1878
/label=U626 (Arabidopsis thaliana)
/note="U626 (Arabidopsis thaliana)"
/regulatory_class="promoter"
misc_RNA 1902..1977
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(1989..2013)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 2031..2232
/label=U626 (Arabidopsis thaliana)
/note="U626 (Arabidopsis thaliana)"
/regulatory_class="promoter"
misc_RNA 2256..2338
/label=sgRNA
/note="sgRNA"
misc_RNA 2256..2331
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(2343..2367)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter 2996..3340
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 3413..7516
/label=Cas9
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
misc_feature 7529..7549
/label=SV40 NLS
/note="SV40 NLS"
CDS 7529..7549
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
terminator 7576..7828
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 8115..8138
/label=RB
/note="RB"
regulatory complement(8203..8243)
/regulatory_class="terminator"
misc_feature 8253..8276
/label=assembly linker
/note="assembly linker"
CDS complement(8290..9102)
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 9204..9227
/label=assembly linker
/note="assembly linker"
CDS complement(9231..9890)
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
rep_origin complement(9891..10662)
/direction=LEFT
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
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