LOCUS       Exported               10709 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gene deletion vector pDGO-01, complete sequence.
ACCESSION   HQ157351
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10709)
  AUTHORS   Olson DG, Tripathi SA, Giannone RJ, Lo J, Caiazza NC, Hogsett DA, 
            Hettich RL, Guss AM, Dubrovsky G, Lynd LR.
  TITLE     Deletion of the Cel48S cellulase from Clostridium thermocellum
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 107 (41), 17727-17732 (2010)
  PUBMED    20837514
REFERENCE   2  (bases 1 to 10709)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-AUG-2010) Thayer School of Engineering, Dartmouth 
            College, 8000 Cummings, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 10709)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10709
                     /organism="Gene deletion vector pDGO-01"
                     /lab_host="Clostridium thermocellum"
                     /mol_type="other DNA"
                     /db_xref="taxon:981225"
     CDS             126..929
                     /codon_start=1
                     /product="URA3"
                     /label=URA3
                     /protein_id="ADW95854.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      1244..1832
                     /label=ColE1
                     /note="ColE1"
     CDS             2524..3528
                     /codon_start=1
                     /product="repB"
                     /label=repB
                     /note="rolling circle replication protein for plasmid 
                     replication in Clostridium thermocellum"
                     /protein_id="ADW95855.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     regulatory      3529..4149
                     /regulatory_class="promoter"
                     /note="cbp; derived from Clostridium thermocellum strain 
                     DSM 1313"
     CDS             4150..5094
                     /codon_start=1
                     /product="pyrF"
                     /label=pyrF
                     /note="negative-selectable marker used with 5FOA"
                     /protein_id="ADW95856.1"
                     /translation="MFIDTLIEKIREKDNPSVVGLDPKIEYVPSFIKEDMYKKYGKNLK
                     AVAEAILLFNKYIIDAVYDIVPAVKPQLAYYEMYGLEGMRVFYETCKYAKEKGLLVIAD
                     GKRNDIGSTAQCYSAAYLGKTDIDEGISEAVFDVDALTVNPYLGVDGIKPFIDDCVKYN
                     KGIFVLVKTSNKSSGEIQDILTQEGRSIYEIVAEYVESWGENKKGKYGYSCVGAVVGAT
                     YPNLAKILRKILKNSYILVPGYGAQGGTARDVAHCFNYDGLGAIVNASRSIMCAYKSEQ
                     WKNVYSEEKFYEASRAEAIRMRDDINSALRDRK"
     terminator      5218..5261
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5393..5420
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5501..5605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5606..6466
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="bla; ampicillin resistance in E. coli"
                     /protein_id="ADW95857.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     protein_bind    6625..6641
                     /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     6649..6665
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(6710..7767)
                     /note="cel48S 3 flanking region; derived from Clostridium 
                     thermocellum strain DSM 1313"
     regulatory      7841..8365
                     /regulatory_class="promoter"
                     /note="gapDH; derived from Clostridium thermocellum strain 
                     DSM 1313"
     CDS             8366..9016
                     /codon_start=1
                     /product="cat"
                     /label=cat
                     /note="provides thiamphenicol resistance in C. 
                     thermocellum"
                     /protein_id="ADW95858.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     misc_feature    complement(9097..10064)
                     /note="cel48S 5 flanking region; derived from Clostridium 
                     thermocellum strain DSM 1313"
     rep_origin      complement(10083..10596)
                     /direction=LEFT
                     /label=CEN/ARSH
                     /note="CEN/ARSH"
     misc_feature    10083..10586
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        10619..125
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter