pHCT2 vector (V005576)

LOCUS       V005576                11112 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005576
VERSION     V005576
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11112)
  AUTHORS   Alexander WG, Peris D, Pfannenstiel BT, Opulente DA, Kuang M,
            Hittinger CT.
  TITLE     Efficient engineering of marker-free synthetic allotetraploids of
            Saccharomyces
  JOURNAL   Fungal Genet. Biol. (2015) In press
   PUBMED   26555931
REFERENCE   2  (bases 1 to 11112)
  AUTHORS   Alexander WG, Peris D, Pfannenstiel B, Opulente DA, Kuang M,
            Hittinger CT.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-SEP-2015) Laboratory of Genetics, University of
            Wisconsin-Madison, 425-G Henry Mall, 4350 Genetics Biotechnology
            Center, Madison, WI 53706, USA
REFERENCE   3  (bases 1 to 11112)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11112)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Fungal
            Genet. Biol. (2015) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (08-SEP-2015) Laboratory of Genetics, University of
            Wisconsin-Madison, 425-G Henry Mall, 4350 Genetics Biotechnology
            Center, Madison, WI 53706, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(31..1035)
                     /label="tTA"
                     /note="tetracycline-controlled transactivator, comprising a
                     fusion of the tetracycline repressor TetR with the
                     C-terminal activation domain of herpes simplex virus VP16"
     promoter        complement(1140..1343)
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(1344..1723)
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     terminator      1835..2022
                     /label="ADH1 terminator"
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     protein_bind    2052..2322
                     /label="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     CDS             2510..4267
                     /gene="HO"
                     /label="Homothallic switching endonuclease"
                     /note="Homothallic switching endonuclease from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P09932"
     misc_feature    4292..4315
                     /gene="HO"
                     /label="6x glycine linker"
                     /note="6x glycine linker"
     CDS             4316..4813
                     /label="13xMyc"
                     /note="13 tandem Myc epitope tags"
     terminator      5006..5253
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5276..5292)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    5300..5316
                     /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(5324..5354)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5369..5390)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5678..6266)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6440..7297)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7298..7402)
                     /label="AmpR promoter"
     gene            7610..8729
                     /label="natMX6"
                     /note="yeast selectable marker conferring nourseothricin
                     resistance"
     rep_origin      9072..9909
                     /label="ARS1"
                     /note="S. cerevisiae autonomously replicating sequence
                     ARS1/ARS416"
     misc_feature    10045..10337
                     /label="CEN4"
                     /note="S. cerevisiae CEN4 centromere"
     primer_bind     11096..11112
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"