LOCUS       Exported                9963 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZJ20, complete sequence.
ACCESSION   KP057688
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9963)
  AUTHORS   Zhou J, Olson DG, Lanahan AA, Tian L, Murphy SJ, Lo J, Lynd LR.
  TITLE     Physiological roles of pyruvate ferredoxin oxidoreductase and 
            pyruvate formate-lyase in Thermoanaerobacterium saccharolyticum 
            JW/SL-YS485
  JOURNAL   Biotechnol Biofuels 8, 138 (2015)
  PUBMED    26379770
REFERENCE   2  (bases 1 to 9963)
  AUTHORS   Zhou J.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-OCT-2014) Thayer School of Engineering, Dartmouth 
            College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9963)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9963
                     /organism="Cloning vector pZJ20"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:1619667"
     oriT            1..825
     oriT            347..456
                     /note="incP origin of transfer"
     regulatory      877..1079
                     /regulatory_class="terminator"
                     /note="T1T2 transcriptional terminator from pEX18"
     terminator      877..904
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      996..1082
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1144..2059
                     /label=upstream of Tsac_0628
                     /note="upstream of Tsac_0628"
     CDS             2209..3267
                     /codon_start=1
                     /gene="pta"
                     /product="phosphate acetyltransferase"
                     /label=pta
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77044.1"
                     /translation="MYTIYFFLVRGMYKNKYFKGRDDLMSIIQNIIEKAKSDKKKIVLP
                     EGAEPRTLKAAEIVLKEGIADLVLLGNEDEIRNAAKDLDISKAEIIDPVKSEMFDRYAN
                     DFYELRKNKGITLEKARETIKDNIYFGCMMVKEGYADGLVSGAIHATADLLRPAFQIIK
                     TAPGAKIVSSFFIMEVPNCEYGENGVFLFADCAVNPSPNAEELASIAVQSANTAKNLLG
                     FEPKVAMLSFSTKGSASHELVDKVRKATEIAKELMPDVAIDGELQLDAALVKEVAELKA
                     PGSKVAGCANVLIFPDLQAGNIGYKLVQRLAKANAIGPITQGMGAPVNDLSRGCSYRDI
                     VDVIATTAVQAQ"
     gene            2209..3267
                     /gene="pta"
                     /label=pta
     CDS             3285..4496
                     /codon_start=1
                     /gene="ack"
                     /product="acetate kinase"
                     /label=ack
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77043.1"
                     /translation="MKIMKILVINCGSSSLKYQLIESTDGNVLAKGLAERIGINDSMLT
                     HNANGEKIKIKKDMKDHKDAIKLVLDALVNSDYGVIKDMSEIDAVGHRVVHGGESFTSS
                     VLINDEVLKAITDCIELAPLHNPANIEGIKACQQIMPNVPMVAVFDTAFHQTMPDYAYL
                     YPIPYEYYTKYRIRRYGFHGTSHKYVSNRAAEILNKPIEDLKIITCHLGNGSSIAAVKY
                     GKSIDTSMGFTPLEGLAMGTRSGSIDPSIISYLMEKENISAEEVVNILNKKSGVYGISG
                     ISSDFRDLEDAAFKNGDERAQLALNVFAYRVKKTIGAYAAAMGGVDVIVFTAGVGENGP
                     EIREFILDGLEFLGFSLDKEKNKVRGKETIISTPNSKVSVMVVPTNEEYMIAKDTEKIV
                     KSIK"
     gene            3285..4496
                     /gene="ack"
                     /label=ack
     regulatory      4624..5154
                     /regulatory_class="promoter"
                     /note="kanamycin promoter"
     CDS             5155..5949
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /note="KanR"
                     /protein_id="AJP77045.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6285..7187
                     /label=downstream of Tsac_0628
                     /note="downstream of Tsac_0628"
     rep_origin      7330..8242
                     /label=p15A origin of replication
                     /note="p15A origin of replication"
     rep_origin      complement(7597..8142)
                     /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        8254..8282
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(8356..9159)
                     /codon_start=1
                     /gene="ura3"
                     /product="Ura3"
                     /label=ura3
                     /note="derived from Saccharomyces cerevisiae"
                     /protein_id="AJP77046.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            complement(8356..9159)
                     /gene="ura3"
                     /label=ura3
     promoter        complement(9160..9375)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     misc_feature    9408..9911
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"