LOCUS       Exported               11734 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEBM101, complete sequence.
ACCESSION   KC879307
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11734)
  AUTHORS   Lynd LR.
  TITLE     Identification of sporulation histidine kinases in C. thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11734)
  AUTHORS   Mearls EB.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-2013) Engineering, Dartmouth College, 14 
            Engineering Dr, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 11734)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11734
                     /organism="Cloning vector pEBM101"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:1346746"
     misc_recomb     1..1165
     regulatory      1213..1466
                     /regulatory_class="terminator"
     terminator      1214..1461
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      1650..2323
     rep_origin      complement(1709..2297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2384..2962)
                     /codon_start=1
                     /gene="tdk"
                     /product="tdk"
                     /label=tdk
                     /protein_id="AGO62066.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     gene            complement(2384..2962)
                     /gene="tdk"
                     /label=tdk
     regulatory      2963..3583
                     /regulatory_class="promoter"
                     /gene="cbp"
     gene            2963..3583
                     /gene="cbp"
                     /label=cbp
     rep_origin      3584..5118
     CDS             complement(3659..4663)
                     /codon_start=1
                     /gene="repB"
                     /product="repB"
                     /label=repB
                     /note="replication protein B"
                     /protein_id="AGO62067.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(3659..4663)
                     /gene="repB"
                     /label=repB
     CDS             complement(5221..6081)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(6177..6980)
                     /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(6981..7196)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      7219..7737
     misc_feature    7229..7732
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_recomb     7893..8815
     misc_recomb     8816..9794
     RBS             9076..9084
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     regulatory      9812..10459
                     /regulatory_class="promoter"
                     /gene="gapD"
     gene            9812..10459
                     /gene="gapD"
                     /label=gapD
     CDS             10460..11110
                     /codon_start=1
                     /gene="cat"
                     /product="cat"
                     /label=cat
                     /note="chloramphenicol acetyl transferase"
                     /protein_id="AGO62068.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            10460..11110
                     /gene="cat"
                     /label=cat
     CDS             11128..11673
                     /codon_start=1
                     /gene="hpt"
                     /product="hpt"
                     /label=hpt
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="AGO62069.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            11128..11673
                     /gene="hpt"
                     /label=hpt