CasYFP vector (V010040)

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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