LOCUS       Exported               10413 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMU1810, complete sequence.
ACCESSION   KC779546
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10413)
  AUTHORS   Mearls EB, Olson DG, Herring CD, Lynd LR.
  TITLE     Development of a regulatable plasmid-based gene expression system 
            for Clostridium thermocellum
  JOURNAL   Appl. Microbiol. Biotechnol. 99 (18), 7589-7599 (2015)
  PUBMED    25994254
REFERENCE   2  (bases 1 to 10413)
  AUTHORS   Argyros A, Mearls EB.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-2013) Engineering, Dartmouth College, 14 
            Engineering Dr, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 10413)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10413
                     /organism="Cloning vector pMU1810"
                     /mol_type="other DNA"
                     /country="USA"
                     /label=derived from Clostridium thermocellum strain BL21
                     /note="derived from Clostridium thermocellum strain BL21"
                     /db_xref="taxon:1381391"
     CDS             complement(291..1094)
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /note="orotidine 5-phosphate decarboxylase"
                     /protein_id="AGT95730.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(291..1094)
                     /gene="ura3"
                     /label=ura3
     promoter        complement(1095..1310)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      1333..1846
     misc_feature    1343..1846
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_recomb     1888..3099
                     /label=spo0A 3 flank for homologous recombination
                     /note="spo0A 3 flank for homologous recombination"
     misc_recomb     3100..4192
                     /label=spo0A 5 flank for homologous recombination
                     /note="spo0A 5 flank for homologous recombination"
     CDS             complement(4311..4856)
                     /codon_start=1
                     /gene="hpt"
                     /product="Hpt"
                     /label=hpt
                     /note="hypoxanthine phosphoribosyl transferase"
                     /protein_id="AGT95731.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            complement(4311..4856)
                     /gene="hpt"
                     /label=hpt
     CDS             complement(4879..5529)
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /note="chloramphenicol acetyltransferase"
                     /protein_id="AGT95732.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            complement(4879..5529)
                     /gene="cat"
                     /label=cat
     regulatory      complement(5530..6057)
                     /regulatory_class="promoter"
                     /gene="gapD"
     gene            complement(5530..6057)
                     /gene="gapD"
                     /label=gapD
     misc_feature    6058..6800
                     /note="fragment of Spo0A; sporulation master regulator-like
                     protein"
     CDS             complement(6801..7379)
                     /codon_start=1
                     /gene="tdk"
                     /product="Tdk"
                     /label=tdk
                     /note="thymidine kinase"
                     /protein_id="AGT95733.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(6801..7379)
                     /gene="tdk"
                     /label=tdk
     regulatory      complement(7380..8000)
                     /regulatory_class="promoter"
                     /gene="cbp"
     gene            complement(7380..8000)
                     /gene="cbp"
                     /label=cbp
     CDS             complement(8107..9111)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /note="replication protein B"
                     /protein_id="AGT95734.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(8107..9111)
                     /gene="repB"
                     /label=repB
     rep_origin      complement(9803..10391)
                     /direction=LEFT