LOCUS       Exported                5500 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMU749, complete sequence.
ACCESSION   JN880474
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5500)
  AUTHORS   Olson DG.
  TITLE     Transformation of Clostridium thermocellum by electroporation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5500)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2011) Thayer School of Engineering, Dartmouth 
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 5500)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5500
                     /organism="Cloning vector pMU749"
                     /mol_type="other DNA"
                     /label=for transformation of Clostridium thermocellum
                     /note="for transformation of Clostridium thermocellum"
                     /db_xref="taxon:1155009"
     rep_origin      1..589
                     /label=eco ori
                     /note="eco ori"
     rep_origin      826..1280
                     /label=cth ori
                     /note="cth ori"
     CDS             1281..2285
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /protein_id="AFC37769.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            1281..2285
                     /gene="repB"
                     /label=repB
     promoter        2400..2504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2505..3365
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AFC37770.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2505..3365
                     /gene="bla"
                     /label=bla
     protein_bind    3524..3540
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3548..3564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3631..4144)
                     /direction=LEFT
                     /label=CEN6/ARSH
                     /note="CEN6/ARSH"
     misc_feature    3631..4134
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4167..4382
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4383..5186
                     /codon_start=1
                     /product="URA3"
                     /label=URA3
                     /protein_id="AFC37771.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"