pSH073 vector (V003246)

LOCUS       V003246                10397 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003246
VERSION     V003246
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10397)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
            Zheng T, Papanek B, Guss AM, Lynd LR.
  TITLE     The ethanol pathway from Thermoanaerobacterium saccharolyticum
            improves ethanol production in Clostridium thermocellum
  JOURNAL   Metab. Eng. 42, 175-184 (2017)
   PUBMED   28663138
REFERENCE   2  (bases 1 to 10397)
  AUTHORS   Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
            Zheng T, Papanek BA, Guss AM, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAR-2017) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 10397)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10397)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            42, 175-184 (2017)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (27-MAR-2017) Thayer School of Engineering, Dartmouth College, 14
            Engineering Drive, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..92
                     /label="cat promoter"
                     /note="cat promoter"
                     /regulatory_class="promoter"
     CDS             93..740
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             complement(824..1681)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(1682..1773)
                     /label="AmpR promoter"
     rep_origin      2033..2578
                     /direction=RIGHT
                     /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             3049..4050
                     /label="repB"
                     /note="RepB replication protein"
     regulatory      4099..4307
                     /label="Clo1313_2638 promoter"
                     /note="Clo1313_2638 promoter"
                     /regulatory_class="promoter"
     CDS             4308..5507
                     /codon_start=1
                     /gene="adhA"
                     /product="AdhA"
                     /label="adhA"
                     /note="NADPH-dependent alcohol dehydrogenase"
                     /protein_id="ASK86096.1"
                     /translation="MWETKVNPSKIFELRCKNTTYFGVGSIHKIKDILENLKINGINNV
                     IFITGKGSYKTSGAWDVVRPVLEELDLKYSLYDKVGPNPTVDMIDEAAKIGRESGAKAV
                     IGIGGGSPIDTAKSVAVLLKYTDKNARELYKQKFIPDDAVPIIAINLTHGTGTEVDRFA
                     VATIPEKNYKPAIAYDCLYPMFAIDDPSLMTKLDKKQTIAVTVDALNHITEAATTLVAS
                     PYSILTAKETVRLIVRYLPAAVNDPLNIVARYYLLYASALAGISFDNGLLHLTHALEHP
                     LSAVKPEIAHGLGLGAILPAVIKAIYPATAEVLADVYSPIVPGLKGLPVEAEYVAEKVQ
                     EWLFSVGCIQKLSDFGFTKDDIPNLVKLAKTTPSLDGLLSIAPVEATESVIEKIYLKSL
                     "
     gene            4308..5507
                     /gene="adhA"
                     /label="adhA"
     CDS             5525..6379
                     /codon_start=1
                     /gene="nfnA"
                     /product="NfnA"
                     /label="nfnA"
                     /note="NADH-dependent reduced ferredoxin:NADP+
                     oxidoreductase"
                     /protein_id="ASK86098.1"
                     /translation="MNEILEKKQLNPTVKMMVINAPLMAKKAKPGQFVIVRVDEKGERI
                     PLTIADYDRNKGTITIIFQEVGMSTKKLGTLNVGDRLHDFVGPLGKPVEFSKDTKRVLA
                     IGGGVGVAPLYPKVKMLNEMKVPVDSIIGGRSAEYVILEDEMKKVSENLYITTDDGTKG
                     RKGFVTDVLKELIEKDNKYDEVIAIGPLIMMKMVCNITKEYNIPTMVSMNPIMIDGTGM
                     CGGCRVTVGGETKFACVDGPAFDGLKVDFDEAMRRQNMYKDMERKVLENYEHECKLGGI
                     LNG"
     gene            5525..6379
                     /gene="nfnA"
                     /label="nfnA"
     CDS             6372..7763
                     /codon_start=1
                     /gene="nfnB"
                     /product="NfnB"
                     /label="nfnB"
                     /note="NADH-dependent reduced ferredoxin:NADP+
                     oxidoreductase"
                     /protein_id="ASK86099.1"
                     /translation="MANMSLKKVPMPEQEPDQRNKNFKEVALGYEENMAVEEAERCIQC
                     KNQPCVEGCPVHVKIPEFIKLIANRDFEGAYQKIKETNNLPAICGRVCPQESQCESVCT
                     RGKKGEPVAIGRLERFTADWHMKNNEDKIEKPETNGRKVAVIGSGPAGLSCAGDLAKMG
                     YDTTIFEAFHTPGGVLMYGIPEFRLPKEIVQKEIDSLKKLGVKIETNMVIGKILTIDDL
                     FDMGYEAVFIGTGAGLPKFMNIPGENLNGVYSANEFLTRINLMKAYDFPNSPTPVKVGK
                     KVAVVGGGNVAMDAARSAKRMGAEEVYIVYRRSEEEMPARLEEIHHAKEEGIIFKLLTN
                     PVRIIGDESGSVKGIECVNMVLGDVDESGRRRPVEEKGSEHVIDVDTVIIAIGQSPNPL
                     ITSTTEGLEKQRWGGIIVNEETLETSRRGVFAGGDAVTGAATVILAMGAGKKAAASIHK
                     YLSEK"
     gene            6372..7763
                     /gene="nfnB"
                     /label="nfnB"
     CDS             7781..10360
                     /codon_start=1
                     /gene="adhE"
                     /product="AdhE"
                     /label="adhE"
                     /note="bifunctional alcohol and aldehyde dehydrogenase"
                     /protein_id="ASK86097.1"
                     /translation="MATTKTELDVQKQIDLLVSRAQEAQKKFMSYTQEQIDAIVKAMAL
                     AGVDKHVELAKMAYEETKMGVYEDKITKNLFATEYVYHDIKNEKTVGIINENIEENYME
                     VAEPIGVIAGVTPVTNPTSTTMFKCLISIKTRNPIIFSFHPKAIKCSIAAAKVMYEAAL
                     KAGAPEGCIGWIETPSIEATQLLMTHPGVSLILATGGAGMVKAAYSSGKPALGVGPGNV
                     PCYIEKSANIKRAVSDLILSKTFDNGVICASEQAVIIDEEIADEVKKLMKEYGCYFLNK
                     DEIKKLEKFAIDEQSCAMSPAVVGQPAAKIAEMAGFKVPEGTKILVAEYEGVGPKYPLS
                     REKLSPILACYTVKDYNEGIKKCEEMTEFGGLGHSAVIHSENQNVINEFARRVRTGRLI
                     VNSPSSQGAIGDIYNTNTPSLTLGCGSMGRNSTTDNVSVKNLLNIKRVVIRKDRMKWFK
                     IPPKIYFESGSLQYLCKVKRKKAFIVTDPFMVKLGFVDKVTYQLDKANIEYEIFSEVEP
                     DPSVDTVMNGVKIMNSYNPDLIIAVGDGSAIDAAKGMWLFYEYPDTEFETLRLKFADIR
                     KRAFKFPELGKKALFIAIPTTSGTGSEVTAFAVITDKKRNIKYPLADYELTPDIAIIDP
                     DLTKTVPPSVTADTGMDVLTHAIEAYVSVMASDYTDALAEKAIKIVFEYLPRAYKNGND
                     EEAREKMHNASCMAGMAFTNAFLGINHSMAHILGGKFHIPHGRANAILLPYVIRYNAEK
                     PTKFVAFPQYEYPKAAERYAEIAKFLGLPASTVEEGVESLIEAIKNLMKELNIPLTLKD
                     AGINKEQFEKEIEEMSDIAFNDQCTGTNPRMPLTKEIAEIYRKAYGA"
     gene            7781..10360
                     /gene="adhE"
                     /label="adhE"