pET-21-Com15_thyA-cat vector (V016287)

LOCUS       V016287                 8192 bp    DNA     circular SYN 20-JUL-2021
DEFINITION  Exported.
ACCESSION   V016287
VERSION     V016287
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8192)
  AUTHORS   Chen V, Griffin ME, Maguin P, Varble A, Hang HC.
  TITLE     RecT recombinase expression enables efficient gene editing in
            Enterococcus
  JOURNAL   Appl Environ Microbiol, AEM0084421 (2021) In press
   PUBMED   34232061
REFERENCE   2  (bases 1 to 8192)
  AUTHORS   Chen V, Griffin ME, Hang HC.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAR-2021) Laboratory of Chemical Biology and Microbial
            Pathogenesis, The Rockefeller University, 1230 York Ave., New York,
            NY 10065, USA
REFERENCE   3  (bases 1 to 8192)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Appl
            Environ Microbiol, AEM0084421 (2021) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-MAR-2021) Laboratory of Chemical Biology and Microbial
            Pathogenesis, The Rockefeller University, 1230 York Ave., New York,
            NY 10065, USA"
FEATURES             Location/Qualifiers
     source          1..8192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /label="AmpR promoter"
     CDS             599..1456
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1630..2218
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2404..2546)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2651..2839)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3614..3635)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3651..4730)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4731..4808)
                     /label="lacI promoter"
     promoter        5117..5135
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5136..5160
                     /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)."
     RBS             5175..5197
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    5204..6247
                     /label="homology arm 1"
                     /note="homology arm 1"
     gene            5204..6193
                     /gene="CVT45_04745"
                     /label="CVT45_04745"
     gene            6204..6247
                     /gene="thyA"
                     /label="thyA"
     regulatory      6278..6330
                     /regulatory_class="promoter"
     regulatory      6288..6293
                     /regulatory_class="minus_35_signal"
     regulatory      6312..6317
                     /regulatory_class="minus_10_signal"
     regulatory      6348..6353
                     /regulatory_class="ribosome_binding_site"
     CDS             6360..7007
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    7035..8034
                     /label="homology arm 2"
                     /note="homology arm 2"
     gene            7035..7938
                     /gene="thyA"
                     /label="thyA"
     gene            7958..8035
                     /gene="CVT45_04755"
                     /label="CVT45_04755"
     CDS             8036..8053
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      8120..8167
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"