Basic Vector Information
- Vector Name:
- CasYFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10437 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP.
- Promoter:
- T3
CasYFP 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 ℃.
CasYFP vector Sequence
LOCUS 40924_435 10437 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector CasYFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10437)
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 10437)
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 10437)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10437)
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..10437
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 4..362
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(385..401)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
misc_RNA complement(785..860)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory complement(881..1380)
/label=U6/C8
/note="U6/C8"
/regulatory_class="promoter"
regulatory 1441..2254
/label=from Chlamydomonas reinhardtii
/note="from Chlamydomonas reinhardtii"
/regulatory_class="promoter"
CDS 2273..2293
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 2291..6394
/label=Cas9
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
CDS 6401..7114
/label=EYFP
/note="enhanced YFP"
CDS 7130..7186
/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 7187..7987
/codon_start=1
/product="aminoglycoside 3'-phosphotransferase VIII"
/function="Selection marker in Chlamydomonas reinhardtii"
/label=aminoglycoside 3'-phosphotransferase VIII
/note="selection marker aminoglycoside
3'-phosphotransferase gene (aph) from Streptomyces rimosus"
/protein_id="AQN67803.1"
/translation="DDALRALRGRYPGCEWVVVEDGASGAGVYRLRGGGRELFVKVAAL
GAGVGLLGEAERLVWLAEVGIPVPRVVEGGGDERVAWLVTEAVPGRPASARWPREQRLD
VAVALAGLARSLHALDWERCPFDRSLAVTVPQAARAVAEGSVDLEDLDEERKGWSGERL
LAELERTRPADEDLAVCHGDLCPDNVLLDPRTCEVTGLIDVGRVGRADRHSDLALVLRE
LAHEEDPWFGPECSAAFLREYGRGWDGAVSEEKLAFYRLLDEFF"
regulatory 7991..8230
/regulatory_class="terminator"
promoter complement(8249..8267)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8288..8304)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8312..8328
/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(8336..8366)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(8381..8402)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8690..9278)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9452..10309)
/label=AmpR
/note="beta-lactamase"
promoter complement(10310..10414)
/label=AmpR promoter
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