CasYFP vector (V010040)

Basic Vector Information

Vector Name:
Antibiotic Resistance:
10437 bp
Cloning vector
Replication origin:
Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP.

CasYFP vector Vector Map

CasYFP10437 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000f1 oriSK primergRNA scaffoldU6/C8from Chlamydomonas reinhardtiiSV40 NLSEYFPP2Aaminoglycoside 3'-phosphotransferase VIIIT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_435       10437 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector CasYFP, complete sequence.
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 
  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)
  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 
     misc_RNA        complement(785..860)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      complement(881..1380)
     regulatory      1441..2254
                     /label=from Chlamydomonas reinhardtii
                     /note="from Chlamydomonas reinhardtii"
     CDS             2273..2293
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2291..6394
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             6401..7114
                     /note="enhanced YFP"
     CDS             7130..7186
                     /product="2A peptide from porcine teschovirus-1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
     CDS             7187..7987
                     /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"
     regulatory      7991..8230
     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 
     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)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     CDS             complement(9452..10309)
     promoter        complement(10310..10414)
                     /label=AmpR promoter

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