LOCUS       40924_48468       10034 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZJ15, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10034)
  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 10034)
  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 10034)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10034)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            Biofuels 8, 138 (2015)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-OCT-2014) Thayer School of Engineering, Dartmouth College, 14 
            Engineering Drive, Hanover, NH 03755, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(347..456)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(877..904)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(996..1082)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    1144..2137
                     /label=upstream of Tsac_0380
                     /note="upstream of Tsac_0380"
     CDS             2287..3345
                     /codon_start=1
                     /gene="pta"
                     /product="phosphate acetyltransferase"
                     /label=pta
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77032.1"
                     /translation="MYTIYFFLVRGMYKNKYFKGRDDLMSIIQNIIEKAKSDKKKIVLP
                     EGAEPRTLKAAEIVLKEGIADLVLLGNEDEIRNAAKDLDISKAEIIDPVKSEMFDRYAN
                     DFYELRKNKGITLEKARETIKDNIYFGCMMVKEGYADGLVSGAIHATADLLRPAFQIIK
                     TAPGAKIVSSFFIMEVPNCEYGENGVFLFADCAVNPSPNAEELASIAVQSANTAKNLLG
                     FEPKVAMLSFSTKGSASHELVDKVRKATEIAKELMPDVAIDGELQLDAALVKEVAELKA
                     PGSKVAGCANVLIFPDLQAGNIGYKLVQRLAKANAIGPITQGMGAPVNDLSRGCSYRDI
                     VDVIATTAVQAQ"
     gene            2287..3345
                     /gene="pta"
                     /label=pta
     CDS             3363..4574
                     /codon_start=1
                     /gene="ack"
                     /product="acetate kinase"
                     /label=ack
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77031.1"
                     /translation="MKIMKILVINCGSSSLKYQLIESTDGNVLAKGLAERIGINDSMLT
                     HNANGEKIKIKKDMKDHKDAIKLVLDALVNSDYGVIKDMSEIDAVGHRVVHGGESFTSS
                     VLINDEVLKAITDCIELAPLHNPANIEGIKACQQIMPNVPMVAVFDTAFHQTMPDYAYL
                     YPIPYEYYTKYRIRRYGFHGTSHKYVSNRAAEILNKPIEDLKIITCHLGNGSSIAAVKY
                     GKSIDTSMGFTPLEGLAMGTRSGSIDPSIISYLMEKENISAEEVVNILNKKSGVYGISG
                     ISSDFRDLEDAAFKNGDERAQLALNVFAYRVKKTIGAYAAAMGGVDVIVFTAGVGENGP
                     EIREFILDGLEFLGFSLDKEKNKVRGKETIISTPNSKVSVMVVPTNEEYMIAKDTEKIV
                     KSIK"
     gene            3363..4574
                     /gene="ack"
                     /label=ack
     regulatory      4702..5232
                     /label=kanamycin promoter
                     /note="kanamycin promoter"
                     /regulatory_class="promoter"
     CDS             5233..6024
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    6363..7258
                     /label=downstream of Tsac_0381
                     /note="downstream of Tsac_0381"
     rep_origin      complement(7668..8213)
                     /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        8325..8353
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             complement(8430..9230)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(9231..9451)
                     /label=URA3 promoter
     misc_feature    9479..9982
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 ctagagtcga tcttcgccag cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc
       61 gggtcgtcgg tgagccagag tttcagcagg ccgcccaggc ggcccaggtc gccattgatg
      121 cgggccagct cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg
      181 acggccagca ggtaggccga caggctcatg ccggccgccg ccgccttttc ctcaatcgct
      241 cttcgttcgt ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg
      301 gtttcatcag ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc
      361 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac
      421 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa
      481 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat
      541 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa gcgctgcttc
      601 cctgctgttt tgtggaatat ctaccgactg gaaacaggca aatgcaggaa attactgaac
      661 tgaggggaca ggcgagagac gatgccaaag agctacaccg acgagctggc cgagtgggtt
      721 gaatcccgcg cggccaagaa gcgccggcgt gatgaggctg cggttgcgtt cctggcggtg
      781 agggcggatg tcgatatgcg taaggagaaa ataccgcatc aggcgcatat ttgaatgtat
      841 ttagaaaaat aaacaaaaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
      901 ttctgcttaa tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
      961 cgttgcttcg caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     1021 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     1081 tgatgcctgg ctcatcgagg tatccaagcg attcaatagt aacagtcctt gtatgccctc
     1141 tttctctgcc gcatctttat tggaaagatg tccatggggc gaaagtatcc ttttctttaa
     1201 aggccatggg tacgtgccgt ttatcagcat gtccacgtcg tggttggatt ctaacagcac
     1261 tacgtcggaa tccattatgt tcgcagcaac acctctactt atatagccta aatctgtagc
     1321 gatggtggct ttattctgac cgcaaaaaaa cgagaaacct acaggctctg acgcatcgtg
     1381 agatttctta aatggccgta tttttacatc gcccaatgtg aagtcttcac ctgtcttgaa
     1441 aaatcgcttg ttgtgtggct caatttcccc tatgaggcct tccatagaaa cccatgttgc
     1501 ctcattggca tatatgggaa tgtcgtacct tctggacaat atgcctgcgc cttttatgtg
     1561 gtcgttgtgc tcatgcgtta tcaaaatcgc tgagatatca tgcggatcta catctatgtt
     1621 tagaaggctt ttttctattg tgctgccgct taaacctgca tccacaagta tatgtacatc
     1681 tttatatcct atatatgcag catttccgct gcttccagat ctaagtgaac aaaacctcat
     1741 aaaataccca aaagcggggc tacccccttc caaatttcat aagtcctcta atattatata
     1801 acaatcaaaa tcctttttct atggcaattt gcagaaagtc tgttttcaag acttatcaac
     1861 atataaaatg tgagcaactt tttttaaatg acctttatag tcgttttttg tctaaaaata
     1921 gtacatttta aatgatactt tttaacacag gacttatcca catatgcaca tttacatgtg
     1981 gataactcat agaatagtca caggcgaatt gtcttagcca ttttcatctg taaaatgtat
     2041 ttatttaaag ataatattcc atttttattg ataatgatta tcggtagtgg tataattgaa
     2101 ttacaagata ttgtaaataa gggggactaa tgtatggtaa accgctaagg catgatagct
     2161 aaagcgtgcc cattgtgaag tggattgtat tctacaatta aacctaatac gctcataata
     2221 tgcgcctttc taaaaaatta ttaattgtac ttattatttt ataaaaaata tgttaaaatg
     2281 taaaatgtgt atacaatata tttcttctta gtaagaggaa tgtataaaaa taaatatttt
     2341 aaaggaaggg acgatcttat gagcattatt caaaacatca ttgaaaaagc taaaagcgat
     2401 aaaaagaaaa ttgttctgcc agaaggtgca gaacccagga cattaaaagc tgctgaaata
     2461 gttttaaaag aagggattgc agatttagtg cttcttggaa atgaagatga gataagaaat
     2521 gctgcaaaag acttggacat atccaaagct gaaatcattg accctgtaaa gtctgaaatg
     2581 tttgataggt atgctaatga tttctatgag ttaaggaaga acaaaggaat cacgttggaa
     2641 aaagccagag aaacaatcaa ggataatatc tattttggat gtatgatggt taaagaaggt
     2701 tatgctgatg gattggtatc tggcgctatt catgctactg cagatttatt aagacctgca
     2761 tttcagataa ttaaaacggc tccaggagca aagatagtat caagcttttt tataatggaa
     2821 gtgcctaatt gtgaatatgg tgaaaatggt gtattcttgt ttgctgattg tgcggtcaac
     2881 ccatcgccta atgcagaaga acttgcttct attgccgtac aatctgctaa tactgcaaag
     2941 aatttgttgg gctttgaacc aaaagttgcc atgctatcat tttctacaaa aggtagtgca
     3001 tcacatgaat tagtagataa agtaagaaaa gcgacagaga tagcaaaaga attgatgcca
     3061 gatgttgcta tcgacggtga attgcaattg gatgctgctc ttgttaaaga agttgcagag
     3121 ctaaaagcgc cgggaagcaa agttgcggga tgtgcaaatg tgcttatatt ccctgattta
     3181 caagctggta atataggata taagcttgta cagaggttag ctaaggcaaa tgcaattgga
     3241 cctataacac aaggaatggg tgcaccggtt aatgatttat caagaggatg cagctataga
     3301 gatattgttg acgtaatagc aacaacagct gtgcaggctc aataaaatgt aaagtatgga
     3361 ggatgaaaat tatgaaaata ctggttatta attgcggaag ttcttcgcta aaatatcaac
     3421 tgattgaatc aactgatgga aatgtgttgg caaaaggcct tgctgaaaga atcggcataa
     3481 atgattccat gttgacacat aatgctaacg gagaaaaaat caagataaaa aaagacatga
     3541 aagatcacaa agacgcaata aaattggttt tagatgcttt ggtaaacagt gactacggcg
     3601 ttataaaaga tatgtctgag atagatgctg taggacatag agttgttcac ggaggagaat
     3661 cttttacatc atcagttctc ataaatgatg aagtgttaaa agcgataaca gattgcatag
     3721 aattagctcc actgcacaat cctgctaata tagaaggaat taaagcttgc cagcaaatca
     3781 tgccaaacgt tccaatggtg gcggtatttg atacagcctt tcatcagaca atgcctgatt
     3841 atgcatatct ttatccaata ccttatgaat actacacaaa gtacaggatt agaagatatg
     3901 gatttcatgg cacatcgcat aaatatgttt caaatagggc tgcagagatt ttgaataaac
     3961 ctattgaaga tttgaaaatc ataacttgtc atcttggaaa tggctccagc attgctgctg
     4021 tcaaatatgg taaatcaatt gacacaagca tgggatttac accattagaa ggtttggcta
     4081 tgggtacacg atctggaagc atagacccat ccatcatttc gtatcttatg gaaaaagaaa
     4141 atataagcgc tgaagaagta gtaaatatat taaataaaaa atctggtgtt tacggtattt
     4201 caggaataag cagcgatttt agagacttag aagatgccgc ctttaaaaat ggagatgaaa
     4261 gagctcagtt ggctttaaat gtgtttgcat atcgagtaaa gaagacgatt ggcgcttatg
     4321 cagcagctat gggaggcgtc gatgtcattg tatttacagc aggtgttggt gaaaatggtc
     4381 ctgagatacg agaatttata cttgatggat tagagttttt agggttcagc ttggataaag
     4441 aaaaaaataa agtcagagga aaagaaacta ttatatctac gccgaattca aaagttagcg
     4501 tgatggttgt gcctactaat gaagaataca tgattgctaa agatactgaa aagattgtaa
     4561 agagtataaa atagcattct tgacaaatgt ttaccccatt agtataatta attttggcaa
     4621 ttatattggg gtgagaaaat gaaaattgat ttatcaaaaa ttaaaggaca taggggccgc
     4681 agcatcgaag tcaactacgt aaaacccagc gaaccatttg aggtgatagg taagattata
     4741 ccgaggtatg aaaacgagaa ttggaccttt acagaattac tctatgaagc gccatattta
     4801 aaaagctacc aagacgaaga ggatgaagag gatgaggagg cagattgcct tgaatatatt
     4861 gacaatactg ataagataat atatctttta tatagaagat atcgccgtat gtaaggattt
     4921 cagggggcaa ggcataggca gcgcgcttat caatatatct atagaatggg caaagcataa
     4981 aaacttgcat ggactaatgc ttgaaaccca ggacaataac cttatagctt gtaaattcta
     5041 tcataattgt ggtttcaaaa tcggctccgt cgatactatg ttatacgcca actttcaaaa
     5101 caactttgaa aaagctgttt tctggtattt aaggttttag aatgcaagga acagtgaatt
     5161 ggagttcgtc ttgttataat tagcttcttg gggtatcttt aaatactgta gaaaagagga
     5221 aggaaataat aaatggctaa aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa
     5281 taccgctgcg taaaagatac ggaaggaatg tctcctgcta aggtatataa gctggtggga
     5341 gaaaatgaaa acctatattt aaaaatgacg gacagccggt ataaagggac cacctatgat
     5401 gtggaacggg aaaaggacat gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc
     5461 ctgcactttg aacggcatga tggctggagc aatctgctca tgagtgaggc cgatggcgtc
     5521 ctttgctcgg aagagtatga agatgaacaa agccctgaaa agattatcga gctgtatgcg
     5581 gagtgcatca ggctctttca ctccatcgac atatcggatt gtccctatac gaatagctta
     5641 gacagccgct tagccgaatt ggattactta ctgaataacg atctggccga tgtggattgc
     5701 gaaaactggg aagaagacac tccatttaaa gatccgcgcg agctgtatga ttttttaaag
     5761 acggaaaagc ccgaagagga acttgtcttt tcccacggcg acctgggaga cagcaacatc
     5821 tttgtgaaag atggcaaagt aagtggcttt attgatcttg ggagaagcgg cagggcggac
     5881 aagtggtatg acattgcctt ctgcgtccgg tcgatcaggg aggatatcgg ggaagaacag
     5941 tatgtcgagc tattttttga cttactgggg atcaagcctg attgggagaa aataaaatat
     6001 tatattttac tggatgaatt gttttagtac ctagatttag atgtctaaaa agctttttag
     6061 acatctaatc ttttctgaag tacatccgca actgtccata ctctgatgtt ttatatcttt
     6121 tctaaaagtt cgctagatag gggtcccgag cgcctacgag gaatttgtat cggatccgca
     6181 agagattata tcgagtgcct ttaagaaggc taaaaattac gaagatgtga tacacaaaaa
     6241 ggcaaaagat tacggcaaaa acataccgga tagtcaagtt aaaggagtat tgaaacagat
     6301 agagattact gccttaaacc atgtagacaa gattgtcgct gctgaaaaga cgatgcagat
     6361 agccgatgat ggcagaaaaa tacttagacg attttagata actaaaaaag gaacttccat
     6421 ttaggaagtt ccttttttta gcttgcaact tcttggtctt caaactggct gttgtaaagc
     6481 ttagcgtaaa atccgccttt ggcaagaagc tctttgtggg tgccttgctc tacaatgtca
     6541 ccgtgatcca tgacaagtat gagatccgca tctctaattg tagaaagcct gtgggcgatg
     6601 atgaaacttg tgcggttttt cataaggtta tccatagcct tctgtattag tatttctgta
     6661 cgagtatcga cggagctggt ggcttcgtcc agtatcagta tcttaggatc tttaagtatg
     6721 gctcttgcta tagttataag ctgtttctgt ccttgcgata cattggatgc ttcttcgttt
     6781 aaaaccatgt tgtatccgtc tggcagcgtg tggacgaagt gatctacgtg tgccgccttt
     6841 gcggctttta tgacttcctc atctgtagca tcaagttttc cataccttat attgtccatt
     6901 atggtgccat tgtaaagcca agtatcctgc agcaccatgc caaacaatga ccttaaatcg
     6961 cttctggtaa attcccttat gtcatatcca tctatcaata tccttcctgc atttacatcg
     7021 taaaacctca tcaagagttt taccatcgta gtcttgccag cacctgttgg tcccactatg
     7081 gctacttttt gacccggctt aatatcggct gagaagtcat ttattattat cttgtcaggt
     7141 ctatatccaa agtgcacatg gtcaaactgc acatggcctt ccacattttc aagtttcaca
     7201 ggatgctctg tttcaggcac ttcttcttcc tcgtctaaga attcaaacac cctttcggag
     7261 ttaggatgtt ggcagacgct tgttgatacg aggcttaaaa ttgctgaggt aatcggtaaa
     7321 ttcgacccgt ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc
     7381 aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg
     7441 atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca
     7501 gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt
     7561 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc
     7621 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc
     7681 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
     7741 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct
     7801 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct
     7861 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt
     7921 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca
     7981 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt
     8041 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc
     8101 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg
     8161 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc
     8221 atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag
     8281 cacctgaagt cagccccata cgatataagt tgtaattctc atgtttgaca gcttatcatc
     8341 gataagcttt aatgcggtag tttatcacag ttaaattgct aacgcagtca ggcacctata
     8401 catgcattta cttataatac agttttttag ttttgctggc cgcatcttct caaatatgct
     8461 tcccagcctg cttttctgta acgttcaccc tctaccttag catcccttcc ctttgcaaat
     8521 agtcctcttc caacaataat aatgtcagat cctgtagaga ccacatcatc cacggttcta
     8581 tactgttgac ccaatgcgtc tcccttgtca tctaaaccca caccgggtgt cataatcaac
     8641 caatcgtaac cttcatctct tccacccatg tctctttgag caataaagcc gataacaaaa
     8701 tctttgtcgc tcttcgcaat gtcaacagta cccttagtat attctccagt agatagggag
     8761 cccttgcatg acaattctgc taacatcaaa aggcctctag gttcctttgt tacttcttct
     8821 gccgcctgct tcaaaccgct aacaatacct gggcccacca caccgtgtgc attcgtaatg
     8881 tctgcccatt ctgctattct gtatacaccc gcagagtact gcaatttgac tgtattacca
     8941 atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt acttggcgga taatgccttt
     9001 agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga tatccacatg tgtttttagt
     9061 aaacaaattt tgggacctaa tgcttcaact aactccagta attccttggt ggtacgaaca
     9121 tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga tattaaatag cttggcagca
     9181 acaggactag gatgagtagc agcacgttcc ttatatgtag ctttcgacat gatttatctt
     9241 cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga atactgggca atttcatgtt
     9301 tcttcaacac tacatatgcg tatatatacc aatctaagtc tgtgctcctt ccttcgttct
     9361 tccttctgtt cggagattac cgaatcaaaa aaatttcaaa gaaaccgaaa tcaaaaaaaa
     9421 gaataaaaaa aaaatgatga attgaattga aaagctagct tatcgatggg tccttttcat
     9481 cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat
     9541 agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac
     9601 tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta
     9661 atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta
     9721 cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa
     9781 tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct
     9841 tcattccgta actcttctac cttctttatt tactttctaa aatccaaata caaaacataa
     9901 aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg
     9961 tgtaagttac aggcaagcga tctctaagaa accattatta tcatgacatt aacctataaa
    10021 aaaggcctct cgag
//
