LOCUS       Exported                6950 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDGO145, complete sequence.
ACCESSION   KY852359
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6950)
  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 6950)
  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 6950)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6950
                     /organism="Cloning vector pDGO145"
                     /mol_type="other DNA"
                     /label=derived from Clostridium thermocellum
                     /note="derived from Clostridium thermocellum"
                     /db_xref="taxon:2018468"
     CDS             complement(1..579)
                     /codon_start=1
                     /gene="tdk"
                     /product="Tdk"
                     /label=tdk
                     /note="thymidine kinase"
                     /protein_id="ASK05960.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(1..579)
                     /gene="tdk"
                     /label=tdk
     regulatory      complement(580..1200)
                     /regulatory_class="promoter"
                     /note="C therm cbp promoter"
     CDS             complement(1276..2280)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /note="replication initiator protein"
                     /protein_id="ASK05961.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(1276..2280)
                     /gene="repB"
                     /label=repB
     rep_origin      complement(2281..2735)
                     /direction=LEFT
                     /label=C thermocellum replication origin
                     /note="C thermocellum replication origin"
     CDS             complement(2838..3698)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="ampicillin resistance"
                     /protein_id="ASK05962.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2838..3698)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(3699..3803)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3896..4470
                     /regulatory_class="promoter"
                     /note="C therm gapDH promoter"
     CDS             4471..5121
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /note="chloramphenicol acetyltransferase"
                     /protein_id="ASK05963.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            4471..5121
                     /gene="cat"
                     /label=cat
     CDS             5139..5684
                     /codon_start=1
                     /gene="hpt"
                     /product="Hpt"
                     /label=hpt
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="ASK05964.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            5139..5684
                     /gene="hpt"
                     /label=hpt
     rep_origin      complement(6112..6939)
                     /direction=LEFT
                     /label=E coli replication origin
                     /note="E coli replication origin"
     rep_origin      6139..6684
                     /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."