pdCas9d.e.f vector (V015649)

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LOCUS       62056_8305        9458 bp DNA     circular SYN 24-NOV-2020
DEFINITION  Expression vector pdCas9d.e.f, complete sequence.
ACCESSION   MW221490
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9458)
  AUTHORS   Alvarez B, Mencia M, de Lorenzo V, Fernandez LA.
  TITLE     In vivo diversification of target genomic sites using processive 
            base deaminase fusions blocked by dCas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9458)
  AUTHORS   Alvarez B, Fernandez LA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-NOV-2020) Microbial Biotechnology, CNB-CSIC (Centro 
            Nacional de Biotecnologia-CSIC), Darwin 3, Cantoblanco, Madrid 
            28049, Spain
REFERENCE   3  (bases 1 to 9458)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-NOV-2020) Microbial Biotechnology, CNB-CSIC (Centro Nacional de 
            Biotecnologia-CSIC), Darwin 3, Cantoblanco, Madrid 28049, Spain"
FEATURES             Location/Qualifiers
     source          1..9458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..660)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(661..763)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(1289..1834)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1946..1974
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     misc_RNA        complement(2292..2370)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     CDS             2666..6769
                     /label=dCas9
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     misc_feature    6793..6924
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   6925..6960
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes
                     CRISPR/Cas system"
     misc_feature    6965..6990
                     /label=d spacer sequence
                     /note="d spacer sequence"
     repeat_region   6991..7026
     misc_feature    7031..7056
                     /label=e spacer sequence
                     /note="e spacer sequence"
     repeat_region   7057..7092
     misc_feature    7097..7122
                     /label=f spacer sequence
                     /note="f spacer sequence"
     repeat_region   7123..7158