pDGO-01 vector (V008010)

LOCUS       V008010                10709 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V008010
VERSION     V008010
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10709)
  AUTHORS   Olson DG, Tripathi SA, Giannone RJ, Lo J, Caiazza NC, Hogsett DA,
            Hettich RL, Guss AM, Dubrovsky G, Lynd LR.
  TITLE     Deletion of the Cel48S cellulase from Clostridium thermocellum
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 107 (41), 17727-17732 (2010)
   PUBMED   20837514
REFERENCE   2  (bases 1 to 10709)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-AUG-2010) Thayer School of Engineering, Dartmouth
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 10709)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10709)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2010"; volume: "107"; issue: "41"; pages:
            "17727-17732"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (18-AUG-2010) Thayer School of Engineering, Dartmouth College, 8000
            Cummings, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             126..926
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     rep_origin      1244..1832
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             2524..3525
                     /label="repB"
                     /note="RepB replication protein"
     regulatory      3529..4149
                     /note="cbp; derived from Clostridium thermocellum strain
                     DSM 1313"
                     /regulatory_class="promoter"
     CDS             4150..5094
                     /codon_start=1
                     /product="pyrF"
                     /label="pyrF"
                     /note="negative-selectable marker used with 5FOA"
                     /protein_id="ADW95856.1"
                     /translation="MFIDTLIEKIREKDNPSVVGLDPKIEYVPSFIKEDMYKKYGKNLK
                     AVAEAILLFNKYIIDAVYDIVPAVKPQLAYYEMYGLEGMRVFYETCKYAKEKGLLVIAD
                     GKRNDIGSTAQCYSAAYLGKTDIDEGISEAVFDVDALTVNPYLGVDGIKPFIDDCVKYN
                     KGIFVLVKTSNKSSGEIQDILTQEGRSIYEIVAEYVESWGENKKGKYGYSCVGAVVGAT
                     YPNLAKILRKILKNSYILVPGYGAQGGTARDVAHCFNYDGLGAIVNASRSIMCAYKSEQ
                     WKNVYSEEKFYEASRAEAIRMRDDINSALRDRK"
     terminator      5220..5301
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5393..5420
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        5501..5605
                     /label="AmpR promoter"
     CDS             5606..6463
                     /label="AmpR"
                     /note="beta-lactamase"
     protein_bind    6625..6641
                     /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     6649..6665
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(6710..7767)
                     /note="cel48S 3' flanking region; derived from Clostridium
                     thermocellum strain DSM 1313"
     regulatory      7841..8365
                     /note="gapDH; derived from Clostridium thermocellum strain
                     DSM 1313"
                     /regulatory_class="promoter"
     CDS             8366..9013
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     misc_feature    complement(9097..10064)
                     /note="cel48S 5' flanking region; derived from Clostridium
                     thermocellum strain DSM 1313"
     misc_feature    complement(10083..10586)
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"