LOCUS       40924_48463       10000 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZJ13, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10000)
  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 10000)
  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 10000)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10000)
  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..10000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(40..67)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(159..245)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    307..1228
                     /label=upstream of Tsac_0046
                     /note="upstream of Tsac_0046"
     CDS             1378..2436
                     /codon_start=1
                     /gene="pta"
                     /product="phosphate acetyltransferase"
                     /label=pta
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77027.1"
                     /translation="MYTIYFFLVRGMYKNKYFKGRDDLMSIIQNIIEKAKSDKKKIVLP
                     EGAEPRTLKAAEIVLKEGIADLVLLGNEDEIRNAAKDLDISKAEIIDPVKSEMFDRYAN
                     DFYELRKNKGITLEKARETIKDNIYFGCMMVKEGYADGLVSGAIHATADLLRPAFQIIK
                     TAPGAKIVSSFFIMEVPNCEYGENGVFLFADCAVNPSPNAEELASIAVQSANTAKNLLG
                     FEPKVAMLSFSTKGSASHELVDKVRKATEIAKELMPDVAIDGELQLDAALVKEVAELKA
                     PGSKVAGCANVLIFPDLQAGNIGYKLVQRLAKANAIGPITQGMGAPVNDLSRGCSYRDI
                     VDVIATTAVQAQ"
     gene            1378..2436
                     /gene="pta"
                     /label=pta
     CDS             2454..3665
                     /codon_start=1
                     /gene="ack"
                     /product="acetate kinase"
                     /label=ack
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77028.1"
                     /translation="MKIMKILVINCGSSSLKYQLIESTDGNVLAKGLAERIGINDSMLT
                     HNANGEKIKIKKDMKDHKDAIKLVLDALVNSDYGVIKDMSEIDAVGHRVVHGGESFTSS
                     VLINDEVLKAITDCIELAPLHNPANIEGIKACQQIMPNVPMVAVFDTAFHQTMPDYAYL
                     YPIPYEYYTKYRIRRYGFHGTSHKYVSNRAAEILNKPIEDLKIITCHLGNGSSIAAVKY
                     GKSIDTSMGFTPLEGLAMGTRSGSIDPSIISYLMEKENISAEEVVNILNKKSGVYGISG
                     ISSDFRDLEDAAFKNGDERAQLALNVFAYRVKKTIGAYAAAMGGVDVIVFTAGVGENGP
                     EIREFILDGLEFLGFSLDKEKNKVRGKETIISTPNSKVSVMVVPTNEEYMIAKDTEKIV
                     KSIK"
     gene            2454..3665
                     /gene="ack"
                     /label=ack
     regulatory      3793..4323
                     /label=kanamycin promoter
                     /note="kanamycin promoter"
                     /regulatory_class="promoter"
     CDS             4324..5115
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    5454..6387
                     /label=downstream of Tsac_0046
                     /note="downstream of Tsac_0046"
     rep_origin      complement(6797..7342)
                     /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        7454..7482
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             complement(7559..8359)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(8360..8580)
                     /label=URA3 promoter
     misc_feature    8608..9111
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     oriT            complement(9510..9619)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 tatttagaaa aataaacaaa aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
       61 gccttctgct taatttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
      121 ggccgttgct tcgcaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
      181 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
      241 atttgatgcc tggctcatcg aggtatccaa gcgattcaat agtaacagtc cttgtatgcc
      301 ctcttttcat gtaaggctca gggtaattta ctgcaaagcc atttttgcac tttgcaacag
      361 tggcttcaac gactttccct tttcctggca cttcgtatga cacttcaaac gtgtcgtttc
      421 cgctgccgcc tatcgtcaaa tctaatagtt cttttctact ttcaggaata attatctctc
      481 ttccacttct taaaatatca gaaacttcat caggtagtat aaagtaatct aaaatgccat
      541 tttccatatt ttctatctca ttcctttcac acgatatgaa aatttttgaa ataatagacg
      601 tgatattatg cagaatttta atatttgaaa tattttataa aagccactat ctatctataa
      661 attagataaa tactggctaa gcggtaaatt tgcaaagcaa ccctttcata gaaaccccct
      721 atttactttc cttcaatgta gaaaaaattc atgtaattta ttttatcatt ttcaagcatt
      781 aaagtcaatc cctgcttaac aaaatacaca attagccgaa atattaaatt atagatgctg
      841 tggcaactta acaatattat atcacataat atgaccagct cataattttt atgaataaat
      901 ttttgtcaaa aaaatatttt tatattttaa ttatctgtga tataattatc gtggaactgt
      961 tatataaagt ctaaaggtat atatataagc attttcagaa aactgttata taatcgctta
     1021 accgctgttg tataacagta catatttgtt tatgctgtat tataatagtt tgcttatata
     1081 tccttagatt taaatataaa tttaaaaaga gaggggaaat acaatgtcga aggttatgaa
     1141 aaccatggat ggcaatacag ccgctgcaca cgtagcatac gcctttacag aagtagctgc
     1201 catctatcct ataactccat catcacctta aaccgctaag gcatgatagc taaagcgtgc
     1261 ccattgtgaa gtggattgta ttctacaatt aaacctaata cgctcataat atgcgccttt
     1321 ctaaaaaatt attaattgta cttattattt tataaaaaat atgttaaaat gtaaaatgtg
     1381 tatacaatat atttcttctt agtaagagga atgtataaaa ataaatattt taaaggaagg
     1441 gacgatctta tgagcattat tcaaaacatc attgaaaaag ctaaaagcga taaaaagaaa
     1501 attgttctgc cagaaggtgc agaacccagg acattaaaag ctgctgaaat agttttaaaa
     1561 gaagggattg cagatttagt gcttcttgga aatgaagatg agataagaaa tgctgcaaaa
     1621 gacttggaca tatccaaagc tgaaatcatt gaccctgtaa agtctgaaat gtttgatagg
     1681 tatgctaatg atttctatga gttaaggaag aacaaaggaa tcacgttgga aaaagccaga
     1741 gaaacaatca aggataatat ctattttgga tgtatgatgg ttaaagaagg ttatgctgat
     1801 ggattggtat ctggcgctat tcatgctact gcagatttat taagacctgc atttcagata
     1861 attaaaacgg ctccaggagc aaagatagta tcaagctttt ttataatgga agtgcctaat
     1921 tgtgaatatg gtgaaaatgg tgtattcttg tttgctgatt gtgcggtcaa cccatcgcct
     1981 aatgcagaag aacttgcttc tattgccgta caatctgcta atactgcaaa gaatttgttg
     2041 ggctttgaac caaaagttgc catgctatca ttttctacaa aaggtagtgc atcacatgaa
     2101 ttagtagata aagtaagaaa agcgacagag atagcaaaag aattgatgcc agatgttgct
     2161 atcgacggtg aattgcaatt ggatgctgct cttgttaaag aagttgcaga gctaaaagcg
     2221 ccgggaagca aagttgcggg atgtgcaaat gtgcttatat tccctgattt acaagctggt
     2281 aatataggat ataagcttgt acagaggtta gctaaggcaa atgcaattgg acctataaca
     2341 caaggaatgg gtgcaccggt taatgattta tcaagaggat gcagctatag agatattgtt
     2401 gacgtaatag caacaacagc tgtgcaggct caataaaatg taaagtatgg aggatgaaaa
     2461 ttatgaaaat actggttatt aattgcggaa gttcttcgct aaaatatcaa ctgattgaat
     2521 caactgatgg aaatgtgttg gcaaaaggcc ttgctgaaag aatcggcata aatgattcca
     2581 tgttgacaca taatgctaac ggagaaaaaa tcaagataaa aaaagacatg aaagatcaca
     2641 aagacgcaat aaaattggtt ttagatgctt tggtaaacag tgactacggc gttataaaag
     2701 atatgtctga gatagatgct gtaggacata gagttgttca cggaggagaa tcttttacat
     2761 catcagttct cataaatgat gaagtgttaa aagcgataac agattgcata gaattagctc
     2821 cactgcacaa tcctgctaat atagaaggaa ttaaagcttg ccagcaaatc atgccaaacg
     2881 ttccaatggt ggcggtattt gatacagcct ttcatcagac aatgcctgat tatgcatatc
     2941 tttatccaat accttatgaa tactacacaa agtacaggat tagaagatat ggatttcatg
     3001 gcacatcgca taaatatgtt tcaaataggg ctgcagagat tttgaataaa cctattgaag
     3061 atttgaaaat cataacttgt catcttggaa atggctccag cattgctgct gtcaaatatg
     3121 gtaaatcaat tgacacaagc atgggattta caccattaga aggtttggct atgggtacac
     3181 gatctggaag catagaccca tccatcattt cgtatcttat ggaaaaagaa aatataagcg
     3241 ctgaagaagt agtaaatata ttaaataaaa aatctggtgt ttacggtatt tcaggaataa
     3301 gcagcgattt tagagactta gaagatgccg cctttaaaaa tggagatgaa agagctcagt
     3361 tggctttaaa tgtgtttgca tatcgagtaa agaagacgat tggcgcttat gcagcagcta
     3421 tgggaggcgt cgatgtcatt gtatttacag caggtgttgg tgaaaatggt cctgagatac
     3481 gagaatttat acttgatgga ttagagtttt tagggttcag cttggataaa gaaaaaaata
     3541 aagtcagagg aaaagaaact attatatcta cgccgaattc aaaagttagc gtgatggttg
     3601 tgcctactaa tgaagaatac atgattgcta aagatactga aaagattgta aagagtataa
     3661 aatagcattc ttgacaaatg tttaccccat tagtataatt aattttggca attatattgg
     3721 ggtgagaaaa tgaaaattga tttatcaaaa attaaaggac ataggggccg cagcatcgaa
     3781 gtcaactacg taaaacccag cgaaccattt gaggtgatag gtaagattat accgaggtat
     3841 gaaaacgaga attggacctt tacagaatta ctctatgaag cgccatattt aaaaagctac
     3901 caagacgaag aggatgaaga ggatgaggag gcagattgcc ttgaatatat tgacaatact
     3961 gataagataa tatatctttt atatagaaga tatcgccgta tgtaaggatt tcagggggca
     4021 aggcataggc agcgcgctta tcaatatatc tatagaatgg gcaaagcata aaaacttgca
     4081 tggactaatg cttgaaaccc aggacaataa ccttatagct tgtaaattct atcataattg
     4141 tggtttcaaa atcggctccg tcgatactat gttatacgcc aactttcaaa acaactttga
     4201 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt
     4261 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa
     4321 taaatggcta aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc
     4381 gtaaaagata cggaaggaat gtctcctgct aaggtatata agctggtggg agaaaatgaa
     4441 aacctatatt taaaaatgac ggacagccgg tataaaggga ccacctatga tgtggaacgg
     4501 gaaaaggaca tgatgctatg gctggaagga aagctgcctg ttccaaaggt cctgcacttt
     4561 gaacggcatg atggctggag caatctgctc atgagtgagg ccgatggcgt cctttgctcg
     4621 gaagagtatg aagatgaaca aagccctgaa aagattatcg agctgtatgc ggagtgcatc
     4681 aggctctttc actccatcga catatcggat tgtccctata cgaatagctt agacagccgc
     4741 ttagccgaat tggattactt actgaataac gatctggccg atgtggattg cgaaaactgg
     4801 gaagaagaca ctccatttaa agatccgcgc gagctgtatg attttttaaa gacggaaaag
     4861 cccgaagagg aacttgtctt ttcccacggc gacctgggag acagcaacat ctttgtgaaa
     4921 gatggcaaag taagtggctt tattgatctt gggagaagcg gcagggcgga caagtggtat
     4981 gacattgcct tctgcgtccg gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag
     5041 ctattttttg acttactggg gatcaagcct gattgggaga aaataaaata ttatatttta
     5101 ctggatgaat tgttttagta cctagattta gatgtctaaa aagcttttta gacatctaat
     5161 cttttctgaa gtacatccgc aactgtccat actctgatgt tttatatctt ttctaaaagt
     5221 tcgctagata ggggtcccga gcgcctacga ggaatttgta tcggatccgc aagagattat
     5281 atcgagtgcc tttaagaagg ctaaaaatta cgaagatgtg atacacaaaa aggcaaaaga
     5341 ttacggcaaa aacataccgg atagtcaagt taaaggagta ttgaaacaga tagagattac
     5401 tgccttaaac catgtagaca agattgtcgc tgctgaaaag acgatgcaga tagtcccaga
     5461 aagagcagaa gctttgtttg aaaaagctga agaacttgca aaagaaaaat acgaaacata
     5521 caagaagctt gctgaacaaa attagcttca ccacaattaa tgagcgaaat tgctttcgct
     5581 cattaatttt ttatgtgtta gttgcctcct tctaaatttt tctcttgttt atctatttaa
     5641 tgttttatat aattaatgtt ataaactaca taaaaggagc ggatgatatg agagaaaaaa
     5701 taaagatggc aaatcctatc gtagagctgg atggcgatga aatgaccaga atcatctggc
     5761 agatgataaa agacacgctt atacatcctt ttgtagaact gaatacagaa tactacgatc
     5821 tgggtcttaa aaatagagat ttgacagacg accaagtgac tgtagatgcc gcctatgcga
     5881 taaaaaagca caaagtaggc gtcaaatgcg caactataac acctaatgct caaagagtcg
     5941 aagaatacaa cttaaaaaag atgtggaaaa gccccaatgg caccataagg gctatattgg
     6001 acggcaccgt ttttaggact ccaatactgg taaacggcat caaacctctc gtaaaaagct
     6061 ggacaaagcc tataacgatt gcgcgtcacg cttatggaga tgtgtacaaa gacgtagaat
     6121 acagggtgga aaaggcagct aaagcagaat tagtagtgac gtacgacgat aattccatcg
     6181 aaaggcagac catacacgag tttgacggtc caggaatttt gatggggatg cacaacatag
     6241 acaagtcaat aaaaagcttc gcaaaagctt gtttcaacta cgcattaagt caaaagcagg
     6301 acctatggtt tgccacaaaa gacaccattt caaaagtata tgaccacact tttaaagaca
     6361 tattcaatga tatgtacgaa agcgagtagt taggatgttg gcagacgctt gttgatacga
     6421 ggcttaaaat tgctgaggta atcggtaaat tcgacccgtc tatcagctgt ccctcctgtt
     6481 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga
     6541 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     6601 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     6661 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     6721 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     6781 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     6841 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6901 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     6961 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     7021 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     7081 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     7141 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     7201 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     7261 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     7321 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat
     7381 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     7441 gtaattctca tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt
     7501 taaattgcta acgcagtcag gcacctatac atgcatttac ttataataca gttttttagt
     7561 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     7621 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     7681 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     7741 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     7801 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     7861 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     7921 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     7981 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     8041 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     8101 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     8161 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     8221 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     8281 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     8341 tatatgtagc tttcgacatg atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt
     8401 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     8461 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     8521 aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa
     8581 aagctagctt atcgatgggt ccttttcatc acgtgctata aaaataatta taatttaaat
     8641 tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat
     8701 agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt
     8761 tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga
     8821 agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc
     8881 tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat
     8941 tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt
     9001 actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat
     9061 tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ctctaagaaa
     9121 ccattattat catgacatta acctataaaa aaggcctctc gagctagagt cgatcttcgc
     9181 cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt cggtgagcca
     9241 gagtttcagc aggccgccca ggcggcccag gtcgccattg atgcgggcca gctcgcggac
     9301 gtgctcatag tccacgacgc ccgtgatttt gtagccctgg ccgacggcca gcaggtaggc
     9361 cgacaggctc atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt cgtctggaag
     9421 gcagtacacc ttgataggtg ggctgccctt cctggttggc ttggtttcat cagccatccg
     9481 cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag gattcccgtt
     9541 gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc
     9601 tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac
     9661 cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat
     9721 aatgaccccg aagcagggtt atgcagcgga aaagcgctgc ttccctgctg ttttgtggaa
     9781 tatctaccga ctggaaacag gcaaatgcag gaaattactg aactgagggg acaggcgaga
     9841 gacgatgcca aagagctaca ccgacgagct ggccgagtgg gttgaatccc gcgcggccaa
     9901 gaagcgccgg cgtgatgagg ctgcggttgc gttcctggcg gtgagggcgg atgtcgatat
     9961 gcgtaaggag aaaataccgc atcaggcgca tatttgaatg
//
