LOCUS       Exported               10782 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMC200, complete sequence.
ACCESSION   KC675188
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10782)
  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 10782)
  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 10782)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10782
                     /organism="Cloning vector pMC200"
                     /mol_type="other DNA"
                     /db_xref="taxon:1300182"
     misc_feature    1..1500
                     /label=similar to xynA upstream region
                     /note="similar to xynA upstream region"
     misc_feature    1501..1541
                     /label=restriction cassette
                     /note="restriction cassette"
     CDS             1736..2722
                     /codon_start=1
                     /product="pta"
                     /label=pta
                     /protein_id="AGG53967.1"
                     /translation="MSIIQNIIEKAKSDKKKIVLPEGAEPRTLKAAEIVLKEGIADLVL
                     LGNEDEIRNAAKDLDISKAEIIDPVKSEMFDRYANDFYELRKNKGITLEKARETIKDNI
                     YFGCMMVKEGYADGLVSGAIHATADLLRPAFQIIKTAPGAKIVSSFFIMEVPNCEYGEN
                     GVFLFADCAVNPSPNAEELASIAVQSANTAKNLLGFEPKVAMLSFSTKGSASHELVDKV
                     RKATEIAKELMPDVAIDGELQLDAALVKEVAELKAPGSKVAGCANVLIFPDLQAGNIGY
                     KLVQRLAKANAIGPITQGMGAPVNDLSRGCSYRDIVDVIATTAVQAQ"
     CDS             2740..3951
                     /codon_start=1
                     /product="ack"
                     /label=ack
                     /protein_id="AGG53968.1"
                     /translation="MKIMKILVINCGSSSLKYQLIESTDGNVLAKGLAERIGINDSMLT
                     HNANGEKIKIKKDMKDHKDAIKLVLDALVNSDYGVIKDMSEIDAVGHRVVHGGESFTSS
                     VLINDEVLKAITDCIELAPLHNPANIEGIKACQQIMPNVPMVAVFDTAFHQTMPDYAYL
                     YPIPYEYYTKYRIRRYGFHGTSHKYVSNRAAEILNKPIEDLKIITCHLGNGSSIAAVKY
                     GKSIDTSMGFTPLEGLAMGTRSGSIDPSIISYLMEKENISAEEVVNILNKKSGVYGISG
                     ISSDFRDLEDAAFKNGDERAQLALNVFAYRVKKTIGAYAAAMGGVDVIVFTAGVGENGP
                     EIREFILDGLEFLGFSLDKEKNKVRGKETIISTPNSKVSVMVVPTNEEYMIAKDTEKIV
                     KSIK"
     CDS             4610..5404
                     /codon_start=1
                     /product="kanamycin resistance"
                     /label=kanamycin resistance
                     /protein_id="AGG53969.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    5550..6987
                     /label=similar to xynA downstream region
                     /note="similar to xynA downstream region"
     rep_origin      7059..7971
                     /label=p15A origin of replication
                     /note="p15A origin of replication"
     rep_origin      complement(7326..7871)
                     /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        7983..8011
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(8085..8888)
                     /codon_start=1
                     /product="ura3"
                     /label=ura3
                     /protein_id="AGG53970.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(8889..9104)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     misc_feature    9137..9640
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     oriT            10039..10148
                     /note="incP origin of transfer"
     regulatory      10569..10771
                     /regulatory_class="terminator"
     terminator      10569..10596
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      10688..10774
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"