pUB-Cas9-@GL1 vector (V002457)

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"