LOCUS       Exported                9033 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEBM118, complete sequence.
ACCESSION   KC879310
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9033)
  AUTHORS   Lynd LR.
  TITLE     Identification of sporulation histidine kinases in C. thermocellum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9033)
  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 9033)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9033
                     /organism="Cloning vector pEBM118"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:1346749"
     regulatory      96..349
                     /regulatory_class="terminator"
     terminator      97..344
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      533..1206
     rep_origin      complement(592..1180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1351..2211)
                     /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(2307..3110)
                     /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(3111..3326)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      3349..3867
     misc_feature    3359..3862
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     rep_origin      4016..5500
     CDS             complement(4091..5095)
                     /codon_start=1
                     /product="RepB"
                     /label=RepB
                     /note="replication protein B"
                     /protein_id="AGO62078.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     regulatory      5581..6228
                     /regulatory_class="promoter"
                     /gene="gapD"
     gene            5581..6228
                     /gene="gapD"
                     /label=gapD
     CDS             6229..6879
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /note="chloramphenicol acetyl transferase"
                     /protein_id="AGO62079.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            6229..6879
                     /gene="cat"
                     /label=cat
     CDS             6895..9033
                     /codon_start=1
                     /product="histidine kinase"
                     /label=histidine kinase
                     /note="from Clostridium thermocellum Clo1313_1973"
                     /protein_id="AGO62080.1"
                     /translation="MKKRRTIGFLVNDIDVSYSNLVYLMTKKAAEKYDCNLIVYEGRSF
                     NNPSMADSQHQISYSFVDNTRLDGIIITSATIACFISDEEFVEFCKRYEGIPLVSLGIV
                     VPHATSILVDNKPGMKSQVEHLIKDHGFKKIVYISGPPTNTDSVERLEAYCEALRENDI
                     EVDDGLIFYGDFMAQSGYDIIKEIIKNGIEYDAVVCANDEMAFGAIKGIRDLKDAYNLD
                     LTQKGVICGFDDSITSRLVKPSLTTVRQPIEEMCFYAVEAILKKIEGEKVEDVVKLPSV
                     LVKRESCGCKNAAEVDFVSDSSLRLVPGQRIHENLQTYSLDELFDCITRTLKLCSIRSC
                     FIFKYCGGTLLYEVDSIFRDDFRLPEDSELIYAYYNDERITIEDSIKYIKTRCIIPDRF
                     FNEDRRFTYLVNPLFFYDEHFGYVCFEVANDDVLNFEPIRGQISNTLNGALMLMERAKM
                     EESLRETERLASLGQLIGGISHNLMTPIMSISGACFGLEDLVHEYMASIEDPSVTVQDH
                     KEIARDMQEWINKLKEYNSYMSNVIATVKSQAVQLNSDAINEFTIEELVDRVKFIKNNN
                     PQIRKAVLNCKIDIDGKTAIRGDISNLAQIIENLLINAVQSYEGMDNASNIVDLHILKE
                     ENMVKCIVRDYGKGIPNEIRDRLFRRMVTTKGKNGTGLSLLLSYSTIKGKFGGELWVES
                     KENEGSAFYVTIPLTDRE"