LOCUS       Exported                5650 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pMU770, complete sequence.
ACCESSION   JF423902
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5650)
  AUTHORS   Olson DG, Lynd LR.
  TITLE     Computational Design and Characterization of a Temperature-sensitive
            Replicon for Gram Positive Thermophiles
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5650)
  AUTHORS   Olson DG, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-FEB-2011) Thayer School of Engineering, Dartmouth 
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 5650)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5650
                     /organism="Expression vector pMU770"
                     /mol_type="other DNA"
                     /db_xref="taxon:1077342"
     rep_origin      1..589
                     /note="Escherichia coli origin of replication derived from 
                     pUC19"
     CDS             1281..2285
                     /codon_start=1
                     /product="RepB"
                     /label=RepB
                     /note="replication protein"
                     /protein_id="AEN69094.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     promoter        2400..2504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2505..3365
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /note="ampicillin resistance gene derived from pUC19"
                     /protein_id="AEN69095.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     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"
     regulatory      complement(3649..3683)
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      3656..3683
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3764..4414)
                     /codon_start=1
                     /product="Cat"
                     /label=Cat
                     /note="chloramphenicol acetyltransferase; provides 
                     resistance to thiamphenicol"
                     /protein_id="AEN69096.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     regulatory      complement(4415..4942)
                     /regulatory_class="promoter"
                     /gene="gapDH"
                     /note="derived from Clostridium thermocellum DSM1313"
     gene            complement(4415..4942)
                     /gene="gapDH"
                     /label=gapDH
                     /note="derived from Clostridium thermocellum DSM1313"
     regulatory      complement(5022..5310)
                     /regulatory_class="terminator"
                     /note="rrnB terminator; derived from Escherichia coli"
     terminator      5049..5076
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5208..5251
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"