LOCUS       Exported               10309 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  CipA deletion vector pDGO-34, complete sequence.
ACCESSION   JX489219
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10309)
  AUTHORS   Waller BH, Olson DG, Currie DH, Guss AM, Lynd LR.
  TITLE     Exchange of type II dockerin-containing subunits of the Clostridium 
            thermocellum cellulosome as revealed by SNAP-tags
  JOURNAL   FEMS Microbiol. Lett. 338 (1), 46-53 (2013)
  PUBMED    23082914
REFERENCE   2  (bases 1 to 10309)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-AUG-2012) Thayer School of Engineering, Dartmouth 
            College, Thayer School of Engineering, Dartmouth College, Hanover, 
            NH 03755, USA
REFERENCE   3  (bases 1 to 10309)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10309
                     /organism="CipA deletion vector pDGO-34"
                     /mol_type="other DNA"
                     /label=Clostridium thermocellum strain DS16
                     /note="Clostridium thermocellum strain DS16"
                     /db_xref="taxon:1238569"
     rep_origin      1..519
                     /label=CEN6/ARSH4 yeast origin of replication
                     /note="CEN6/ARSH4 yeast origin of replication"
     misc_feature    11..514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_feature    680..1214
                     /label=5 flank (CipA + promoter)
                     /note="5 flank (CipA + promoter)"
     misc_feature    1215..1753
                     /label=3 flank
                     /note="3 flank"
     regulatory      1766..2413
                     /regulatory_class="promoter"
                     /note="gapD promoter from Clostridium thermocellum"
     CDS             2414..3064
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AFU51885.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            2414..3064
                     /gene="cat"
                     /label=cat
     CDS             3082..3627
                     /codon_start=1
                     /gene="hpt"
                     /product="putative hypoxanthine phosphoribosyltransferase"
                     /EC_number="2.4.2.8"
                     /label=hpt
                     /protein_id="AFU51886.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            3082..3627
                     /gene="hpt"
                     /label=hpt
     regulatory      4304..4557
                     /regulatory_class="terminator"
                     /note="CYC1"
     terminator      4305..4552
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      4741..5414
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      complement(4800..5388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5475..6053)
                     /codon_start=1
                     /gene="tdk"
                     /product="thymidine kinase"
                     /EC_number="2.7.1.21"
                     /label=tdk
                     /protein_id="AFU51887.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(5475..6053)
                     /gene="tdk"
                     /label=tdk
     regulatory      complement(6054..6674)
                     /regulatory_class="promoter"
                     /note="CBP; cellulose-binding protein promoter"
     rep_origin      complement(6675..8209)
                     /direction=LEFT
                     /label=putative minimal Clostridium thermocellum ori
                     /note="putative minimal Clostridium thermocellum ori"
     CDS             complement(6750..7754)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein"
                     /label=repB
                     /note="rolling circle replication initiation protein; 
                     Bacillus origin of replication from Geobacillus 
                     stearothermophilus plasmid pTHT15"
                     /protein_id="AFU51884.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(6750..7754)
                     /gene="repB"
                     /label=repB
     CDS             complement(8312..9172)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR; ampiciliin-resistance protein"
                     /protein_id="AFU51888.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(8312..9172)
                     /gene="bla"
                     /label=bla
     CDS             complement(9268..10071)
                     /codon_start=1
                     /gene="ura3"
                     /product="URA3"
                     /label=ura3
                     /note="from yeast; results in ura+, 5-FOA sensitive when 
                     present"
                     /protein_id="AFU51889.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(9268..10071)
                     /gene="ura3"
                     /label=ura3
     promoter        complement(10072..10287)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter