LOCUS       Exported               10707 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEKH-rsgI6, complete sequence.
ACCESSION   KT362976
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10707)
  AUTHORS   Sand A, Holwerda EK, Ruppertsberger NM, Maloney M, Olson DG, Nataf 
            Y, Borovok I, Sonenshein AL, Bayer EA, Lamed R, Lynd LR, Shoham Y.
  TITLE     Three cellulosomal xylanase genes in Clostridium thermocellum are 
            regulated by both vegetative SigA (sigma(A)) and alternative SigI6 
            (sigma(I6)) factors
  JOURNAL   FEBS Lett. 589 (20), 3133-3140 (2015)
  PUBMED    26320414
REFERENCE   2  (bases 1 to 10707)
  AUTHORS   Holwerda EK.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2015) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 10707)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10707
                     /organism="Cloning vector pEKH-rsgI6"
                     /mol_type="other DNA"
                     /db_xref="taxon:1698430"
     misc_feature    11..514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             3262..3807
                     /codon_start=1
                     /product="hypoxanthine phosphoribosyltransferase"
                     /EC_number="2.4.2.8"
                     /label=hypoxanthine phosphoribosyltransferase
                     /protein_id="ALI61571.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     terminator      4703..4950
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(5198..5786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5873..6451)
                     /codon_start=1
                     /product="thymidine kinase"
                     /EC_number="2.7.1.21"
                     /label=thymidine kinase
                     /protein_id="ALI61572.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     CDS             complement(7148..8152)
                     /codon_start=1
                     /product="repB"
                     /EC_number="3.6.4.12"
                     /label=repB
                     /note="rolling circle replication initiation protein"
                     /protein_id="ALI61573.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             complement(8710..9570)
                     /codon_start=1
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=beta-lactamase
                     /protein_id="ALI61574.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(9666..10469)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(10470..10685)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter