LOCUS       Exported                8248 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYD14, complete sequence.
ACCESSION   KF806700
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8248)
  AUTHORS   Deng Y, Olson DG, Zhou J, Herring CD, Joe Shaw A, Lynd LR.
  TITLE     Redirecting carbon flux through exogenous pyruvate kinase to achieve
            high ethanol yields in Clostridium thermocellum
  JOURNAL   Metab. Eng. 15, 151-158 (2013)
  PUBMED    23202749
REFERENCE   2  (bases 1 to 8248)
  AUTHORS   Zhou J, Olson DG, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-NOV-2013) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, U.S
REFERENCE   3  (bases 1 to 8248)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8248
                     /organism="Cloning vector pYD14"
                     /mol_type="other DNA"
                     /db_xref="taxon:1454265"
     rep_origin      complement(1..455)
                     /direction=LEFT
                     /label=Clostridium thermocellum
                     /note="Clostridium thermocellum"
     CDS             complement(558..1418)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AHH35088.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(558..1418)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(1419..1523)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1613..2012
                     /label=similar to Clo1313_1879
                     /note="similar to Clo1313_1879"
     regulatory      2016..2590
                     /regulatory_class="promoter"
                     /gene="GAPDH"
     gene            2016..2590
                     /gene="GAPDH"
                     /label=GAPDH
     CDS             2591..3241
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AHH35086.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            2591..3241
                     /gene="cat"
                     /label=cat
     misc_feature    2989..4390
                     /label=similar to Clo1313_1879
                     /note="similar to Clo1313_1879"
     CDS             3259..3804
                     /codon_start=1
                     /locus_tag="or0940"
                     /product="hypoxanthine phosphoribosyl transferase"
                     /EC_number="2.4.2.8"
                     /label=or0940
                     /protein_id="AHH35087.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            3259..3804
                     /locus_tag="or0940"
                     /label=or0940
     regulatory      3819..3858
                     /regulatory_class="terminator"
                     /note="hairpin"
     misc_feature    4391..4813
                     /label=similar to Clo1313_1879
                     /note="similar to Clo1313_1879"
     regulatory      4847..5051
                     /regulatory_class="terminator"
                     /note="CYC1"
     rep_origin      5235..5908
                     /label=Escherichia coli
                     /note="Escherichia coli"
     rep_origin      complement(5294..5882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5969..6547)
                     /codon_start=1
                     /gene="tdk"
                     /locus_tag="or0326"
                     /product="thymidine kinase"
                     /EC_number="2.7.1.21"
                     /label=tdk
                     /protein_id="AHH35090.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(5969..6547)
                     /gene="tdk"
                     /locus_tag="or0326"
                     /label=tdk
     regulatory      complement(6548..7168)
                     /regulatory_class="promoter"
                     /gene="cbp"
     gene            complement(6548..7168)
                     /gene="cbp"
                     /label=cbp
     CDS             complement(7244..8248)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein"
                     /label=repB
                     /protein_id="AHH35089.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(7244..8248)
                     /gene="repB"
                     /label=repB