pSH121 vector (V003239)

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LOCUS       V003239                12396 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003239
VERSION     V003239
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12396)
  AUTHORS   Hon S, Holwerda EK, Worthen RS, Maloney MI, Tian L, Cui J, Lin PP,
            Lynd LR, Olson DG.
  TITLE     Expressing the Thermoanaerobacterium saccharolyticum pforA in
            engineered Clostridium thermocellum improves ethanol production
  JOURNAL   Biotechnol Biofuels 11, 242 (2018)
   PUBMED   30202437
REFERENCE   2  (bases 1 to 12396)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Worthen RS, Maloney MI, Tian L, Cui J,
            Lin PP, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-APR-2018) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 12396)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12396)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol
            Biofuels 11, 242 (2018)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (20-APR-2018) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..573
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(840..1418)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label="tdk"
                     /protein_id="AYA22162.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(840..1418)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(1419..2039)
                     /label="C. thermocellum cbp promoter"
                     /note="C. thermocellum cbp promoter"
                     /regulatory_class="promoter"
     CDS             complement(2118..3119)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(3120..3574)
                     /direction=LEFT
                     /label="C. thermocellum origin of replication"
                     /note="C. thermocellum origin of replication"
     CDS             complement(3680..4537)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(4538..4642)
                     /label="AmpR promoter"
     misc_feature    4732..5266
                     /label="upstream homology recombination flank"
                     /note="upstream homology recombination flank"
     regulatory      5267..5529
                     /label="T. saccharolyticum pforA promoter"
                     /note="T. saccharolyticum pforA promoter"
                     /regulatory_class="promoter"
     CDS             5530..9057
                     /codon_start=1
                     /gene="pforA"
                     /product="pyruvate ferredoxin oxidoreductase"
                     /label="pforA"
                     /note="derived from Thermoanaerobacterium saccharolyticum"
                     /protein_id="AYA22165.1"
                     /translation="MSKVMKTMDGNTAAAHVAYAFTEVAAIYPITPSSPMAEHVDEWSA
                     HGRKNLFGQEVKVIEMQSEAGAAGAVHGSLAAGALTTTFTASQGLLLMIPNMYKIAGEL
                     LPGVFHVSARALASHALSIFGDHQDVMACRQTGFALLASGSVQEVMDLGSVAHLAAIKG
                     RVPFLHFFDGFRTSHEYQKIEVMDYEDLRKLLDMDAVREFKKRALNPEHPVTRGTAQNP
                     DIYFQEREASNRYYNAVPEIVEEYMKEISKITGREYKLFNYYGAPDAERIVIAMGSVTE
                     TIEETIDYLLKKGEKVGVVKVHLYRPFSFKHFMDAIPKTVKKIAVLDRTKEAGAFGEPL
                     YEDVRAAFYDSEMKPIIVGGRYGLGSKDTTPAQIVAVFDNLKSDTPKNNFTIGIVDDVT
                     YTSLPVGEEIETTAEGTISCKFWGFGSDGTVGANKSAIQIIGDNTDMYAQAYFSYDSKK
                     SGGVTISHLRFGKKPIRSTYLINNADFVACHKQAYVYNYDVLAGLKKGGTFLLNCTWKP
                     EELDEKLPASMKRYIAKNNINFYIINAVDIAKELGLGARINMIMQSAFFKLANIIPIDE
                     AVKHLKDAIVKSYGHKGEKIVNMNYAAVDRGIDALVKVDVPASWANAEDEAKVERNVPD
                     FIKNIADVMNRQEGDKLPVSAFVGMEDGTFPMGTAAYEKRGIAVDVPEWQIDNCIQCNQ
                     CAYVCPHAAIRPFLLNEEEVKNAPEGFTSKKAIGKGLEGLNFRIQVSVLDCTGCGVCAN
                     TCPSKEKSLIMKPLETQLDQAKNWEYAMSLSYKENPLGTDTVKGSQFEKPLLEFSGACA
                     GCGETPYARLVTQLFGDRMLIANATGCSSIWGGSAPSTPYTVNKDGHGPAWANSLFEDN
                     AEFGFGMALAVKQQREKLADIVKEALELDLTQDLKNALKLWLDNFNSSEITKKTANIIV
                     SLIQDYKTDDSKVKELLNEILDRKEYLVKKSQWIFGGDGWAYDIGFGGLDHVLASGEDV
                     NVLVFDTEVYSNTGGQSSKATPVGAIAQFAAAGKGIGKKDLGRIAMSYGYVYVAQIAMG
                     ANQAQTIKALKEAESYPGPSLIIAYAPCINHGIKLGMGCSQIEEKKAVEAGYWHLYRYN
                     PMLKAEGKNPFILDSKAPTASYKEFIMGEVRYSSLAKTFPERAEALFEKAEELAKEKYE
                     TYKKLAEQN"
     gene            5530..9057
                     /gene="pforA"
                     /label="pforA"
                     /note="Tsac_0046"
     misc_feature    9058..9617
                     /label="first downstream homology recombination flank"
                     /note="first downstream homology recombination flank"
     regulatory      9621..10195
                     /label="C. thermocellum gapDH promoter"
                     /note="C. thermocellum gapDH promoter"
                     /regulatory_class="promoter"
     CDS             10196..10843
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             10864..11409
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label="hpt"
                     /protein_id="AYA22167.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            10864..11409
                     /gene="hpt"
                     /label="hpt"
     misc_feature    11594..12153
                     /label="second downstream homology recombination flank"
                     /note="second downstream homology recombination flank"
     terminator      12186..12374
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"