LOCUS       Exported                8975 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMU2051, complete sequence.
ACCESSION   KC146550
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8975)
  AUTHORS   Zhou J, Olson DG, Argyros DA, Deng Y, van Gulik WM, van Dijken JP, 
            Lynd LR.
  TITLE     Atypical Glycolysis in Clostridium thermocellum
  JOURNAL   Appl. Environ. Microbiol. 79 (9), 3000-3008 (2013)
  PUBMED    23435896
REFERENCE   2  (bases 1 to 8975)
  AUTHORS   Argyros A.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-NOV-2012) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 8975)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8975
                     /organism="Cloning vector pMU2051"
                     /mol_type="other DNA"
                     /db_xref="taxon:1288312"
     CDS             complement(291..1094)
                     /codon_start=1
                     /product="ura3"
                     /label=ura3
                     /note="negative selection marker"
                     /protein_id="AGE45854.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1095..1310)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      1333..1846
                     /label=yeast replication origin
                     /note="yeast replication origin"
     misc_feature    1343..1846
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_feature    1888..2407
                     /label=similar to pyruvate phosphate dikinase
                     /note="similar to pyruvate phosphate dikinase"
     misc_feature    2408..2929
                     /label=similar to pyruvate phosphate dikinase
                     /note="similar to pyruvate phosphate dikinase"
     regulatory      complement(2932..3063)
                     /regulatory_class="terminator"
                     /note="hpt terminator"
     CDS             complement(3064..3618)
                     /codon_start=1
                     /product="hypoxanthine phosporibosyl transferase"
                     /label=hypoxanthine phosporibosyl transferase
                     /protein_id="AGE45855.1"
                     /translation="MINQIKEILVTREELKNNAKELGKRISSDYEGKELVLIGVLKGGV
                     VFFADLIREITIPIDVDFISVSSYGNSTKSSGVVRIIKDIDIDITNKHVLIVEDLVDTG
                     LTLHYLKSMFEARGPKDVKICTALDKPSRRKVDLEIDYKGITIPDKFVVGYGLDYAEKY
                     RNLPDVCVLDSSVYTDKEDMD"
     CDS             complement(3634..4284)
                     /codon_start=1
                     /product="cat"
                     /label=cat
                     /protein_id="AGE45856.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     regulatory      complement(4265..4812)
                     /regulatory_class="promoter"
                     /note="glyceraldehyde 3-phosphate dehydrogenase promoter"
     misc_feature    4813..5361
                     /label=similar to pyruvate phosphate dikinase
                     /note="similar to pyruvate phosphate dikinase"
     CDS             complement(5362..5940)
                     /codon_start=1
                     /product="thymidine kinase"
                     /label=thymidine kinase
                     /protein_id="AGE45857.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     regulatory      complement(5941..6561)
                     /regulatory_class="promoter"
                     /note="cbp promoter"
     CDS             complement(6668..7672)
                     /codon_start=1
                     /product="repB"
                     /label=repB
                     /note="replication protein; derived from Clostridium 
                     thermocellum"
                     /protein_id="AGE45858.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     rep_origin      complement(7673..8133)
                     /direction=LEFT
                     /label=Clostridium thermocellum replication origin
                     /note="Clostridium thermocellum replication origin"
     rep_origin      complement(8364..8952)
                     /direction=LEFT
                     /label=Escherichia coli replication origin
                     /note="Escherichia coli replication origin"