LOCUS       Exported               13012 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMC212, complete sequence.
ACCESSION   KC675189
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Currie DH, Herring CD, Guss AM, Olson DG, Hogsett DA, Lynd LR.
  TITLE     Functional heterologous expression of an engineered full length CipA
            from Clostridium thermocellum in Thermoanaerobacterium 
            saccharolyticum
  JOURNAL   Biotechnol Biofuels 6 (1), 32 (2013)
  PUBMED    23448319
REFERENCE   2  (bases 1 to 13012)
  AUTHORS   Currie DH, Herring CD, Guss AM, Olson DG, Hogsett DA, Lynd LR.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2013) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 13012)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13012
                     /organism="Cloning vector pMC212"
                     /mol_type="other DNA"
                     /db_xref="taxon:1300183"
     misc_feature    1..1500
                     /label=similar to xynA upstream region
                     /note="similar to xynA upstream region"
     CDS             1501..3771
                     /codon_start=1
                     /product="CelS HisX10"
                     /label=CelS HisX10
                     /note="family 48 glycoside hydrolase"
                     /protein_id="AGG53971.1"
                     /translation="MVKSRKISILLAVAMLVSIMIPTTAFAGPTKAPTKDGTSYKDLFL
                     ELYGKIKDPKNGYFSPDEGIPYHSIETLIVEAPDYGHVTTSEAFSYYVWLEAMYGNLTG
                     NWSGVETAWKVMEDWIIPDSTEQPGMSSYNPNSPATYADEYEDPSYYPSELKFDTVRVG
                     SDPVHNDLVSAYGPNMYLMHWLMDVDNWYGFGTGTRATFINTFQRGEQESTWETIPHPS
                     IEEFKYGGPNGFLDLFTKDRSYAKQWRYTNAPDAEGRAIQAVYWANKWAKEQGKGSAVA
                     SVVSKAAKMGDFLRNDMFDKYFMKIGAQDKTPATGYDSAHYLMAWYTAWGGGIGASWAW
                     KIGCSHAHFGYQNPFQGWVSATQSDFAPKSSNGKRDWTTSYKRQLEFYQWLQSAEGGIA
                     GGATNSWNGRYEKYPAGTSTFYGMAYVPHPVYADPGSNQWFGFQAWSMQRVMEYYLETG
                     DSSVKNLIKKWVDWVMSEIKLYDDGTFAIPSDLEWSGQPDTWTGTYTGNPNLHVRVTSY
                     GTDLGVAGSLANALATYAAATERWEGKLDTKARDMAAELVNRAWYNFYCSEGKGVVTEE
                     ARADYKRFFEQEVYVPAGWSGTMPNGDKIQPGIKFIDIRTKYRQDPYYDIVYQAYLRGE
                     APVLNYHRFWHEVDLAVAMGVLATYFPDMTYKVPGTPSTKLYGDVNDDGKVNSTDAVAL
                     KRYVLRSGISINTDNADLNEDGRVNSTDLGILKRYILKEIDTLPYKNGGGTGHHHHHHH
                     HHH"
     CDS             3739..3768
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3966..4952
                     /codon_start=1
                     /product="pta"
                     /label=pta
                     /protein_id="AGG53972.1"
                     /translation="MSIIQNIIEKAKSDKKKIVLPEGAEPRTLKAAEIVLKEGIADLVL
                     LGNEDEIRNAAKDLDISKAEIIDPVKSEMFDRYANDFYELRKNKGITLEKARETIKDNI
                     YFGCMMVKEGYADGLVSGAIHATADLLRPAFQIIKTAPGAKIVSSFFIMEVPNCEYGEN
                     GVFLFADCAVNPSPNAEELASIAVQSANTAKNLLGFEPKVAMLSFSTKGSASHELVDKV
                     RKATEIAKELMPDVAIDGELQLDAALVKEVAELKAPGSKVAGCANVLIFPDLQAGNIGY
                     KLVQRLAKANAIGPITQGMGAPVNDLSRGCSYRDIVDVIATTAVQAQ"
     CDS             4970..6181
                     /codon_start=1
                     /product="ack"
                     /label=ack
                     /protein_id="AGG53973.1"
                     /translation="MKIMKILVINCGSSSLKYQLIESTDGNVLAKGLAERIGINDSMLT
                     HNANGEKIKIKKDMKDHKDAIKLVLDALVNSDYGVIKDMSEIDAVGHRVVHGGESFTSS
                     VLINDEVLKAITDCIELAPLHNPANIEGIKACQQIMPNVPMVAVFDTAFHQTMPDYAYL
                     YPIPYEYYTKYRIRRYGFHGTSHKYVSNRAAEILNKPIEDLKIITCHLGNGSSIAAVKY
                     GKSIDTSMGFTPLEGLAMGTRSGSIDPSIISYLMEKENISAEEVVNILNKKSGVYGISG
                     ISSDFRDLEDAAFKNGDERAQLALNVFAYRVKKTIGAYAAAMGGVDVIVFTAGVGENGP
                     EIREFILDGLEFLGFSLDKEKNKVRGKETIISTPNSKVSVMVVPTNEEYMIAKDTEKIV
                     KSIK"
     CDS             6840..7634
                     /codon_start=1
                     /product="kanamycin resistance"
                     /label=kanamycin resistance
                     /protein_id="AGG53974.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7780..9217
                     /label=similar to xynA downstream region
                     /note="similar to xynA downstream region"
     rep_origin      9289..10201
                     /label=p15A origin of replication
                     /note="p15A origin of replication"
     rep_origin      complement(9556..10101)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        10213..10241
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(10315..11118)
                     /codon_start=1
                     /product="ura3"
                     /label=ura3
                     /protein_id="AGG53975.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(11119..11334)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     misc_feature    11367..11870
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     oriT            12269..12378
                     /note="incP origin of transfer"
     regulatory      12799..13001
                     /regulatory_class="terminator"
     terminator      12799..12826
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      12918..13004
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"