LOCUS       Exported               12613 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEBM100, complete sequence.
ACCESSION   KC879306
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12613)
  AUTHORS   Lynd LR.
  TITLE     Identification of sporulation histidine kinases in C. thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12613)
  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 12613)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12613
                     /organism="Cloning vector pEBM100"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:1346745"
     CDS             1..2139
                     /codon_start=1
                     /product="histidine kinase"
                     /label=histidine kinase
                     /note="from Clostridium thermocellum Clo1313_1973"
                     /protein_id="AGO62061.1"
                     /translation="MKKRRTIGFLVNDIDVSYSNLVYLMTKKAAEKYDCNLIVYEGRSF
                     NNPSMADSQHQISYSFVDNTRLDGIIITSATIACFISDEEFVEFCKRYEGIPLVSLGIV
                     VPHATSILVDNKPGMKSQVEHLIKDHGFKKIVYISGPPTNTDSVERLEAYCEALRENDI
                     EVDDGLIFYGDFMAQSGYDIIKEIIKNGIEYDAVVCANDEMAFGAIKGIRDLKDAYNLD
                     LTQKGVICGFDDSITSRLVKPSLTTVRQPIEEMCFYAVEAILKKIEGEKVEDVVKLPSV
                     LVKRESCGCKNAAEVDFVSDSSLRLVPGQRIHENLQTYSLDELFDCITRTLKLCSIRSC
                     FIFKYCGGTLLYEVDSIFRDDFRLPEDSELIYAYYNDERITIEDSIKYIKTRCIIPDRF
                     FNEDRRFTYLVNPLFFYDEHFGYVCFEVANDDVLNFEPIRGQISNTLNGALMLMERAKM
                     EESLRETERLASLGQLIGGISHNLMTPIMSISGACFGLEDLVHEYMASIEDPSVTVQDH
                     KEIARDMQEWINKLKEYNSYMSNVIATVKSQAVQLNSDAINEFTIEELVDRVKFIKNNN
                     PQIRKAVLNCKIDIDGKTAIRGDISNLAQIIENLLINAVQSYEGMDNASNIVDLHILKE
                     ENMVKCIVRDYGKGIPNEIRDRLFRRMVTTKGKNGTGLSLLLSYSTIKGKFGGELWVES
                     KENEGSAFYVTIPLTDRE"
     regulatory      2188..2441
                     /regulatory_class="terminator"
     terminator      2189..2436
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      2625..3298
     rep_origin      complement(2684..3272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3359..3937)
                     /codon_start=1
                     /product="tdk"
                     /label=tdk
                     /protein_id="AGO62062.1"
                     /translation="MYGPKDHGYIEVVTGPMFSGKSEELIRRIKRAKIARQKVQVFKPA
                     IDDRYSIDKVVSHNGDNMHAIAIVKASDILAYAEEDTDVFAIDEVQFFDSEIVDIVKEI
                     ADSGKRVICAGLDMDFRGEPFGPTPELMAIAEFVDKLTAICMKCGNPATRTQRLINGKP
                     ANYDDPIIMVGAKESYEARCRKCHEVPRT"
     regulatory      3938..4558
                     /regulatory_class="promoter"
                     /note="cbp promoter"
     CDS             complement(4634..5638)
                     /codon_start=1
                     /gene="repB"
                     /product="repB"
                     /label=repB
                     /note="replication protein B"
                     /protein_id="AGO62063.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            complement(4634..5638)
                     /gene="repB"
                     /label=repB
     CDS             complement(6196..7056)
                     /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(7152..7955)
                     /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(7956..8171)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      8194..8712
     misc_feature    8204..8707
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_recomb     8868..9809
     misc_recomb     9810..10673
     regulatory      10691..11338
                     /regulatory_class="promoter"
                     /gene="gapD"
     gene            10691..11338
                     /gene="gapD"
                     /label=gapD
     CDS             11339..11989
                     /codon_start=1
                     /gene="cat"
                     /product="cat"
                     /label=cat
                     /note="chloramphenicol acetyl transferase"
                     /protein_id="AGO62064.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            11339..11989
                     /gene="cat"
                     /label=cat
     CDS             12007..12552
                     /codon_start=1
                     /gene="hpt"
                     /product="hpt"
                     /label=hpt
                     /note="hypoxanthine phosphoribosyltransferase"
                     /protein_id="AGO62065.1"
                     /translation="MENLSKDIDEILITEEELKEKIKELGRQITKDYKGKNLMLVGVLK
                     GALMFMADLSRHIDLPLSLDFMAVSSYGSSTHSSGIVKIIKDLDISIEGKDVLIVEDII
                     DSGLTLSYLRETLLGRKPKSLKICTILDKPERREASVKVDYVGFKIPDKFVVGYGLDFD
                     EKYRNLPFIGVLKPEMYS"
     gene            12007..12552
                     /gene="hpt"
                     /label=hpt