pXJ065 vector (V016307)

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LOCUS       V016307                 8785 bp    DNA     circular SYN 16-MAY-2020
DEFINITION  Exported.
ACCESSION   V016307
VERSION     V016307
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8785)
  AUTHORS   Wang Q.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-FEB-2020) Single-Cell Center, Qingdao Institute of
            Bioenergy and Bioprocess Technology, No. 189 Songling Road, Laoshan
            District, Qingdao, Shandong Province, Qindao, Shandong 266000, China
REFERENCE   2  (bases 1 to 8785)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (20-FEB-2020) Single-Cell Center, Qingdao Institute of Bioenergy and
            Bioprocess Technology, No. 189 Songling Road, Laoshan District,
            Qingdao, Shandong Province, Qindao, Shandong 266000, China"
FEATURES             Location/Qualifiers
     source          1..8785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             1564..2559
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     CDS             2560..2577
                     /label="6xHis"
                     /note="6xHis affinity tag"
     misc_RNA        3260..3335
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     CDS             complement(3623..3655)
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             complement(3659..3676)
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             complement(3686..3703)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     RBS             complement(3722..3744)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(3759..3783)
                     /label="lac operator"
                     /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(3784..3802)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4111..4188
                     /label="lacI promoter"
     CDS             4189..5268
                     /label="lacI"
                     /note="lac repressor"
     protein_bind    5284..5305
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             6080..6268
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    6373..6515
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6701..7289)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             7411..8223
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(8319..8774)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"