pHMK34 vector (V005530)

Basic Vector Information

Vector Name:
pHMK34
Antibiotic Resistance:
Ampicillin
Length:
11568 bp
Type:
Yeast integrative vector
Replication origin:
ori
Source/Author:
Alexander WG, Peris D, Pfannenstiel BT, Opulente DA, Kuang M, Hittinger CT.
Promoter:
CMV

pHMK34 vector Vector Map

pHMK3411568 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500tTACMV promoterADH1 terminatortetracycline response elementHomothallic switching endonuclease13xMycCYC1 terminatorM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterhphMX6ARS1CEN4M13 fwd

pHMK34 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V005530                11568 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005530
VERSION     V005530
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11568)
  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 11568)
  AUTHORS   Alexander WG, Peris D, Pfannenstiel B, Opulente DA, Kuang M,
            Hittinger CT.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-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 11568)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11568)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal
            Genet. Biol. (2015) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-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..11568
                     /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"
     regulatory      complement(1334..1723)
                     /regulatory_class="enhancer"
     enhancer        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"
     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..9185
                     /label="hphMX6"
                     /note="yeast selectable marker conferring hygromycin
                     resistance"
     rep_origin      9528..9981
                     /label="ARS1"
                     /note="S. cerevisiae autonomously replicating sequence
                     ARS1"
     misc_feature    10501..10793
                     /label="CEN4"
                     /note="S. cerevisiae CEN4 centromere"
     primer_bind     11552..11568
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"

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