pCRISPR-Cas9 vector (V008167)

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LOCUS       V008167                11265 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008167
VERSION     V008167
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11265)
  AUTHORS   Tong Y, Charusanti P, Zhang L, Weber T, Lee SY.
  TITLE     CRISPR-Cas9 based engineering of actinomycetal genomes
  JOURNAL   ACS Synth Biol (2015) In press
   PUBMED   25806970
REFERENCE   2  (bases 1 to 11265)
  AUTHORS   Tong Y, Charusanti P, Zhang L, Weber T, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-2015) The Novo Nordisk Foundation Center for
            Biosustainability, Kogle Alle 6, Horsholm 2970, Denmark
REFERENCE   3  (bases 1 to 11265)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11265)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
            Biol (2015) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (23-MAR-2015) The Novo Nordisk Foundation Center for
            Biosustainability, Kogle Alle 6, Horsholm 2970, Denmark"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(467..576)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             892..1692
                     /label="ApmR"
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     rep_origin      1994..2582
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2870..2891
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2906..2936
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2944..2960
                     /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)."
     primer_bind     2968..2984
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_RNA        3123..3198
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     terminator      complement(3368..3402)
                     /label="lambda t0 terminator"
                     /note="minimal transcription terminator from phage lambda
                     (Scholtissek and Grosse, 1987)"
     CDS             3570..7673
                     /label="Cas9"
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     terminator      complement(7863..7911)
                     /label="fd terminator"
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             8255..9061
                     /gene="tsnR"
                     /label="23S rRNA (adenosine(1067)-2'-O)-methyltransferase"
                     /note="23S rRNA (adenosine(1067)-2'-O)-methyltransferase
                     from Streptomyces azureus. Accession#: P18644"
     CDS             9627..11066
                     /label="pSG5 Rep"
                     /note="replication initiator protein from the Streptomyces
                     ghanaensis plasmid pSG5"