LOCUS       Exported                9473 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDGO134, complete sequence.
ACCESSION   KU682277
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9473)
  AUTHORS   Olson DG.
  TITLE     Conversion of phosphoenolpyruvate to pyruvate in Clostridium 
            thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9473)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2016) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9473)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9473
                     /organism="Cloning vector pDGO134"
                     /mol_type="other DNA"
                     /db_xref="taxon:1889895"
     CDS             complement(1..1005)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein"
                     /label=repB
                     /protein_id="AOC59188.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(1..1005)
                     /gene="repB"
                     /label=repB
     rep_origin      complement(1006..1460)
                     /direction=LEFT
                     /label=C. thermocellum replication origin
                     /note="C. thermocellum replication origin"
     CDS             complement(1563..2423)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="Bla; ampicillin resistance protein; provides 
                     antibiotic resistance to ampicillin and carbenicillin"
                     /protein_id="AOC59189.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1563..2423)
                     /gene="bla"
                     /label=bla
     promoter        complement(2424..2528)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4857..5507
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="Cat; provides antibiotic resistance to 
                     thiamphenicol"
                     /protein_id="AOC59191.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            4857..5507
                     /gene="cat"
                     /label=cat
     CDS             5525..6070
                     /codon_start=1
                     /gene="hpt"
                     /product="hypoxanthine phosphoribosyltransferase"
                     /label=hpt
                     /note="Hpt; counterselection with 8AZH"
                     /protein_id="AOC59192.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            5525..6070
                     /gene="hpt"
                     /label=hpt
     misc_feature    6255..7043
                     /note="partial pyruvate kinase gene from T. 
                     saccharolyticum"
     rep_origin      7465..8138
                     /label=E. coli origin of replication
                     /note="E. coli origin of replication"
     rep_origin      complement(7524..8112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8199..8777)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /label=tdk
                     /note="Tdk; counterselection with FUDR"
                     /protein_id="AOC59190.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(8199..8777)
                     /gene="tdk"
                     /label=tdk