pSH074 vector (V003245)

LOCUS       V003245                14612 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003245
VERSION     V003245
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14612)
  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 14612)
  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 (28-MAR-2017) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 14612)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 14612)
  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
            (28-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..14612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..579)
                     /codon_start=1
                     /gene="tdk"
                     /product="Tdk"
                     /label="tdk"
                     /note="thymidine kinase"
                     /protein_id="ASK05943.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(1..579)
                     /gene="tdk"
                     /label="tdk"
     regulatory      complement(580..1200)
                     /label="C therm cbp promoter"
                     /note="C therm cbp promoter"
                     /regulatory_class="promoter"
     CDS             complement(1279..2280)
                     /label="repB"
                     /note="RepB replication protein"
     rep_origin      complement(2281..2735)
                     /direction=LEFT
                     /label="C thermocellum replication origin"
                     /note="C thermocellum replication origin"
     CDS             complement(2841..3698)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(3699..3803)
                     /label="AmpR promoter"
     misc_feature    3893..4392
                     /label="upstream homology region"
                     /note="upstream homology region"
     regulatory      4396..4970
                     /label="C thermocellum gapDH promoter"
                     /note="C thermocellum gapDH promoter"
                     /regulatory_class="promoter"
     CDS             4971..5618
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             5639..6184
                     /codon_start=1
                     /gene="hpt"
                     /product="Hpt"
                     /label="hpt"
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="ASK05947.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            5639..6184
                     /gene="hpt"
                     /label="hpt"
     misc_feature    6369..6868
                     /label="upstream homology region"
                     /note="upstream homology region"
     CDS             6869..8068
                     /codon_start=1
                     /gene="adhA"
                     /product="AdhA"
                     /label="adhA"
                     /note="NADPH-dependent alcohol dehydrogenase"
                     /protein_id="ASK05948.1"
                     /translation="MWETKVNPSKIFELRCKNTTYFGVGSIHKIKDILENLKINGINNV
                     IFITGKGSYKTSGAWDVVRPVLEELDLKYSLYDKVGPNPTVDMIDEAAKIGRESGAKAV
                     IGIGGGSPIDTAKSVAVLLKYTDKNARELYKQKFIPDDAVPIIAINLTHGTGTEVDRFA
                     VATIPEKNYKPAIAYDCLYPMFAIDDPSLMTKLDKKQTIAVTVDALNHITEAATTLVAS
                     PYSILTAKETVRLIVRYLPAAVNDPLNIVARYYLLYASALAGISFDNGLLHLTHALEHP
                     LSAVKPEIAHGLGLGAILPAVIKAIYPATAEVLADVYSPIVPGLKGLPVEAEYVAEKVQ
                     EWLFSVGCIQKLSDFGFTKDDIPNLVKLAKTTPSLDGLLSIAPVEATESVIEKIYLKSL
                     "
     gene            6869..8068
                     /gene="adhA"
                     /label="adhA"
     CDS             8086..8940
                     /codon_start=1
                     /gene="nfnA"
                     /product="NfnA"
                     /label="nfnA"
                     /note="NADH-dependent reduced ferredoxin:NADP+
                     oxidoreductase"
                     /protein_id="ASK05949.1"
                     /translation="MNEILEKKQLNPTVKMMVINAPLMAKKAKPGQFVIVRVDEKGERI
                     PLTIADYDRNKGTITIIFQEVGMSTKKLGTLNVGDRLHDFVGPLGKPVEFSKDTKRVLA
                     IGGGVGVAPLYPKVKMLNEMKVPVDSIIGGRSAEYVILEDEMKKVSENLYITTDDGTKG
                     RKGFVTDVLKELIEKDNKYDEVIAIGPLIMMKMVCNITKEYNIPTMVSMNPIMIDGTGM
                     CGGCRVTVGGETKFACVDGPAFDGLKVDFDEAMRRQNMYKDMERKVLENYEHECKLGGI
                     LNG"
     gene            8086..8940
                     /gene="nfnA"
                     /label="nfnA"
     CDS             8933..10324
                     /codon_start=1
                     /gene="nfnB"
                     /product="NfnB"
                     /label="nfnB"
                     /note="NADH-dependent reduced ferredoxin:NADP+
                     oxidoreductase"
                     /protein_id="ASK05950.1"
                     /translation="MANMSLKKVPMPEQEPDQRNKNFKEVALGYEENMAVEEAERCIQC
                     KNQPCVEGCPVHVKIPEFIKLIANRDFEGAYQKIKETNNLPAICGRVCPQESQCESVCT
                     RGKKGEPVAIGRLERFTADWHMKNNEDKIEKPETNGRKVAVIGSGPAGLSCAGDLAKMG
                     YDTTIFEAFHTPGGVLMYGIPEFRLPKEIVQKEIDSLKKLGVKIETNMVIGKILTIDDL
                     FDMGYEAVFIGTGAGLPKFMNIPGENLNGVYSANEFLTRINLMKAYDFPNSPTPVKVGK
                     KVAVVGGGNVAMDAARSAKRMGAEEVYIVYRRSEEEMPARLEEIHHAKEEGIIFKLLTN
                     PVRIIGDESGSVKGIECVNMVLGDVDESGRRRPVEEKGSEHVIDVDTVIIAIGQSPNPL
                     ITSTTEGLEKQRWGGIIVNEETLETSRRGVFAGGDAVTGAATVILAMGAGKKAAASIHK
                     YLSEK"
     gene            8933..10324
                     /gene="nfnB"
                     /label="nfnB"
     CDS             10342..12921
                     /codon_start=1
                     /gene="adhE"
                     /product="AdhE"
                     /label="adhE"
                     /note="bifunctional alcohol and aldehyde dehydrogenase;
                     bifunctional alcohol and aldehyde dehydrognase"
                     /protein_id="ASK05951.1"
                     /translation="MATTKTELDVQKQIDLLVSRAQEAQKKFMSYTQEQIDAIVKAMAL
                     AGVDKHVELAKMAYEETKMGVYEDKITKNLFATEYVYHDIKNEKTVGIINENIEENYME
                     VAEPIGVIAGVTPVTNPTSTTMFKCLISIKTRNPIIFSFHPKAIKCSIAAAKVMYEAAL
                     KAGAPEGCIGWIETPSIEATQLLMTHPGVSLILATGGAGMVKAAYSSGKPALGVGPGNV
                     PCYIEKSANIKRAVSDLILSKTFDNGVICASEQAVIIDEEIADEVKKLMKEYGCYFLNK
                     DEIKKLEKFAIDEQSCAMSPAVVGQPAAKIAEMAGFKVPEGTKILVAEYEGVGPKYPLS
                     REKLSPILACYTVKDYNEGIKKCEEMTEFGGLGHSAVIHSENQNVINEFARRVRTGRLI
                     VNSPSSQGAIGDIYNTNTPSLTLGCGSMGRNSTTDNVSVKNLLNIKRVVIRKDRMKWFK
                     IPPKIYFESGSLQYLCKVKRKKAFIVTDPFMVKLGFVDKVTYQLDKANIEYEIFSEVEP
                     DPSVDTVMNGVKIMNSYNPDLIIAVGDGSAIDAAKGMWLFYEYPDTEFETLRLKFADIR
                     KRAFKFPELGKKALFIAIPTTSGTGSEVTAFAVITDKKRNIKYPLADYELTPDIAIIDP
                     DLTKTVPPSVTADTGMDVLTHAIEAYVSVMASDYTDALAEKAIKIVFEYLPRAYKNGND
                     EEAREKMHNASCMAGMAFTNAFLGINHSMAHILGGKFHIPHGRANAILLPYVIRYNAEK
                     PTKFVAFPQYEYPKAAERYAEIAKFLGLPASTVEEGVESLIEAIKNLMKELNIPLTLKD
                     AGINKEQFEKEIEEMSDIAFNDQCTGTNPRMPLTKEIAEIYRKAYGA"
     gene            10342..12921
                     /gene="adhE"
                     /label="adhE"
     misc_feature    12939..13457
                     /label="downstream homology region"
                     /note="downstream homology region"
     terminator      13490..13678
                     /label="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      complement(13938..14526)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"