LOCUS       40924_48478       10010 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pZJ17, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10010)
  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 10010)
  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 10010)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10010)
  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..10010
                     /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..2069
                     /label=upstream of Tsac_2177
                     /note="upstream of Tsac_2177"
     CDS             2219..3277
                     /codon_start=1
                     /gene="pta"
                     /product="phosphate acetyltransferase"
                     /label=pta
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77040.1"
                     /translation="MYTIYFFLVRGMYKNKYFKGRDDLMSIIQNIIEKAKSDKKKIVLP
                     EGAEPRTLKAAEIVLKEGIADLVLLGNEDEIRNAAKDLDISKAEIIDPVKSEMFDRYAN
                     DFYELRKNKGITLEKARETIKDNIYFGCMMVKEGYADGLVSGAIHATADLLRPAFQIIK
                     TAPGAKIVSSFFIMEVPNCEYGENGVFLFADCAVNPSPNAEELASIAVQSANTAKNLLG
                     FEPKVAMLSFSTKGSASHELVDKVRKATEIAKELMPDVAIDGELQLDAALVKEVAELKA
                     PGSKVAGCANVLIFPDLQAGNIGYKLVQRLAKANAIGPITQGMGAPVNDLSRGCSYRDI
                     VDVIATTAVQAQ"
     gene            2219..3277
                     /gene="pta"
                     /label=pta
     CDS             3295..4506
                     /codon_start=1
                     /gene="ack"
                     /product="acetate kinase"
                     /label=ack
                     /note="negative selective marker in Thermoanaerobacter 
                     saccharolyticum"
                     /protein_id="AJP77039.1"
                     /translation="MKIMKILVINCGSSSLKYQLIESTDGNVLAKGLAERIGINDSMLT
                     HNANGEKIKIKKDMKDHKDAIKLVLDALVNSDYGVIKDMSEIDAVGHRVVHGGESFTSS
                     VLINDEVLKAITDCIELAPLHNPANIEGIKACQQIMPNVPMVAVFDTAFHQTMPDYAYL
                     YPIPYEYYTKYRIRRYGFHGTSHKYVSNRAAEILNKPIEDLKIITCHLGNGSSIAAVKY
                     GKSIDTSMGFTPLEGLAMGTRSGSIDPSIISYLMEKENISAEEVVNILNKKSGVYGISG
                     ISSDFRDLEDAAFKNGDERAQLALNVFAYRVKKTIGAYAAAMGGVDVIVFTAGVGENGP
                     EIREFILDGLEFLGFSLDKEKNKVRGKETIISTPNSKVSVMVVPTNEEYMIAKDTEKIV
                     KSIK"
     gene            3295..4506
                     /gene="ack"
                     /label=ack
     regulatory      4634..5164
                     /label=kanamycin promoter
                     /note="kanamycin promoter"
                     /regulatory_class="promoter"
     CDS             5165..5956
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    6295..7234
                     /label=downstream of Tsac_2180
                     /note="downstream of Tsac_2180"
     rep_origin      complement(7644..8189)
                     /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        8301..8329
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             complement(8406..9206)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(9207..9427)
                     /label=URA3 promoter
     misc_feature    9455..9958
                     /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 tttccgacca tatagaaaaa ccgcctgata aaagtttgta tatgccgcct gtaagcatac
     1201 cagacaatac tgtcataaat cccgaaccgc cttgattgct tgttacaagg acttcagaag
     1261 ccgccattcc ttctggatac agaagtttcc cgtgttcttc aaccgtcaaa taccttctta
     1321 gaggtacaac aaacaatacg ccaaataatc caccgattaa taccgcaaat attattcttt
     1381 cgagagtaaa ttcattttta aaaccccaaa tagctatagc tggaaaagaa aacaaaagtc
     1441 cagcagcaac actctcacct gttgcggcta ttgctgtagc catattagct tctaatatgc
     1501 tatttcttct aaaaatcgtc tttaatatac cggtagacaa aacagcagca ggaatagctg
     1561 cacttatagt cattccagtt ttaagaccta aataagtatt agctgccgca aaaacaattg
     1621 caaatatcac gcctataact aaactcaaaa atgtagattc tggaagaacc tcttccgccg
     1681 gaacaaatgg cacatacttg tcgccactta taccgccata agcgccatct gatagctttc
     1741 ttccatgttc aacttttcca ctggccatat cttttctcct ttcgtgttat agtttaaata
     1801 aattatttaa gctttttatt accgtaacca cctcctttca atattttaac ttatattaaa
     1861 atattttaat agtgtaaagt aataaaatgg ttacatggtt ttttaacctt ttacaaaacc
     1921 atatatcgtt ttccactttt attatattcg acggacattt caaaaatcct tcaaaaaatg
     1981 caaaatatgc ttcatgaata ttttcattgc atttctttca ttcatgatgt aattttatct
     2041 gttgcttaaa atctctataa gctctgctct aaaccgctaa ggcatgatag ctaaagcgtg
     2101 cccattgtga agtggattgt attctacaat taaacctaat acgctcataa tatgcgcctt
     2161 tctaaaaaat tattaattgt acttattatt ttataaaaaa tatgttaaaa tgtaaaatgt
     2221 gtatacaata tatttcttct tagtaagagg aatgtataaa aataaatatt ttaaaggaag
     2281 ggacgatctt atgagcatta ttcaaaacat cattgaaaaa gctaaaagcg ataaaaagaa
     2341 aattgttctg ccagaaggtg cagaacccag gacattaaaa gctgctgaaa tagttttaaa
     2401 agaagggatt gcagatttag tgcttcttgg aaatgaagat gagataagaa atgctgcaaa
     2461 agacttggac atatccaaag ctgaaatcat tgaccctgta aagtctgaaa tgtttgatag
     2521 gtatgctaat gatttctatg agttaaggaa gaacaaagga atcacgttgg aaaaagccag
     2581 agaaacaatc aaggataata tctattttgg atgtatgatg gttaaagaag gttatgctga
     2641 tggattggta tctggcgcta ttcatgctac tgcagattta ttaagacctg catttcagat
     2701 aattaaaacg gctccaggag caaagatagt atcaagcttt tttataatgg aagtgcctaa
     2761 ttgtgaatat ggtgaaaatg gtgtattctt gtttgctgat tgtgcggtca acccatcgcc
     2821 taatgcagaa gaacttgctt ctattgccgt acaatctgct aatactgcaa agaatttgtt
     2881 gggctttgaa ccaaaagttg ccatgctatc attttctaca aaaggtagtg catcacatga
     2941 attagtagat aaagtaagaa aagcgacaga gatagcaaaa gaattgatgc cagatgttgc
     3001 tatcgacggt gaattgcaat tggatgctgc tcttgttaaa gaagttgcag agctaaaagc
     3061 gccgggaagc aaagttgcgg gatgtgcaaa tgtgcttata ttccctgatt tacaagctgg
     3121 taatatagga tataagcttg tacagaggtt agctaaggca aatgcaattg gacctataac
     3181 acaaggaatg ggtgcaccgg ttaatgattt atcaagagga tgcagctata gagatattgt
     3241 tgacgtaata gcaacaacag ctgtgcaggc tcaataaaat gtaaagtatg gaggatgaaa
     3301 attatgaaaa tactggttat taattgcgga agttcttcgc taaaatatca actgattgaa
     3361 tcaactgatg gaaatgtgtt ggcaaaaggc cttgctgaaa gaatcggcat aaatgattcc
     3421 atgttgacac ataatgctaa cggagaaaaa atcaagataa aaaaagacat gaaagatcac
     3481 aaagacgcaa taaaattggt tttagatgct ttggtaaaca gtgactacgg cgttataaaa
     3541 gatatgtctg agatagatgc tgtaggacat agagttgttc acggaggaga atcttttaca
     3601 tcatcagttc tcataaatga tgaagtgtta aaagcgataa cagattgcat agaattagct
     3661 ccactgcaca atcctgctaa tatagaagga attaaagctt gccagcaaat catgccaaac
     3721 gttccaatgg tggcggtatt tgatacagcc tttcatcaga caatgcctga ttatgcatat
     3781 ctttatccaa taccttatga atactacaca aagtacagga ttagaagata tggatttcat
     3841 ggcacatcgc ataaatatgt ttcaaatagg gctgcagaga ttttgaataa acctattgaa
     3901 gatttgaaaa tcataacttg tcatcttgga aatggctcca gcattgctgc tgtcaaatat
     3961 ggtaaatcaa ttgacacaag catgggattt acaccattag aaggtttggc tatgggtaca
     4021 cgatctggaa gcatagaccc atccatcatt tcgtatctta tggaaaaaga aaatataagc
     4081 gctgaagaag tagtaaatat attaaataaa aaatctggtg tttacggtat ttcaggaata
     4141 agcagcgatt ttagagactt agaagatgcc gcctttaaaa atggagatga aagagctcag
     4201 ttggctttaa atgtgtttgc atatcgagta aagaagacga ttggcgctta tgcagcagct
     4261 atgggaggcg tcgatgtcat tgtatttaca gcaggtgttg gtgaaaatgg tcctgagata
     4321 cgagaattta tacttgatgg attagagttt ttagggttca gcttggataa agaaaaaaat
     4381 aaagtcagag gaaaagaaac tattatatct acgccgaatt caaaagttag cgtgatggtt
     4441 gtgcctacta atgaagaata catgattgct aaagatactg aaaagattgt aaagagtata
     4501 aaatagcatt cttgacaaat gtttacccca ttagtataat taattttggc aattatattg
     4561 gggtgagaaa atgaaaattg atttatcaaa aattaaagga cataggggcc gcagcatcga
     4621 agtcaactac gtaaaaccca gcgaaccatt tgaggtgata ggtaagatta taccgaggta
     4681 tgaaaacgag aattggacct ttacagaatt actctatgaa gcgccatatt taaaaagcta
     4741 ccaagacgaa gaggatgaag aggatgagga ggcagattgc cttgaatata ttgacaatac
     4801 tgataagata atatatcttt tatatagaag atatcgccgt atgtaaggat ttcagggggc
     4861 aaggcatagg cagcgcgctt atcaatatat ctatagaatg ggcaaagcat aaaaacttgc
     4921 atggactaat gcttgaaacc caggacaata accttatagc ttgtaaattc tatcataatt
     4981 gtggtttcaa aatcggctcc gtcgatacta tgttatacgc caactttcaa aacaactttg
     5041 aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg
     5101 tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata
     5161 ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg
     5221 cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga
     5281 aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg
     5341 ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt
     5401 tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc
     5461 ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat
     5521 caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg
     5581 cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg
     5641 ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa
     5701 gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa
     5761 agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta
     5821 tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga
     5881 gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt
     5941 actggatgaa ttgttttagt acctagattt agatgtctaa aaagcttttt agacatctaa
     6001 tcttttctga agtacatccg caactgtcca tactctgatg ttttatatct tttctaaaag
     6061 ttcgctagat aggggtcccg agcgcctacg aggaatttgt atcggatccg caagagatta
     6121 tatcgagtgc ctttaagaag gctaaaaatt acgaagatgt gatacacaaa aaggcaaaag
     6181 attacggcaa aaacataccg gatagtcaag ttaaaggagt attgaaacag atagagatta
     6241 ctgccttaaa ccatgtagac aagattgtcg ctgctgaaaa gacgatgcag atagcgcata
     6301 gttcacagct tttgcacaag ttttcgttaa atatgacttg cttcaatttc atcatctcca
     6361 tcatatttta ttacctcatt cataaacttt tgcttcttct tcgccgctta agacgcggta
     6421 tgctccttca gcaagtgctt gcatttcatc ttctcctgga tacacgtaca caggagcgat
     6481 aaattcgact ctaattgaaa tgcgctttac aaaattttca ttgtatgcta tgccacctgt
     6541 tatacctata gcatccactt cgccatttaa agctacagcc atcgctccaa tctcttttgc
     6601 tgtcttatac gccattgcat caagtattaa ttcagcattc tcatcaccat tttctatcat
     6661 tctgtatact tcttttacgt catttgtatt aagatgtgct actaaaccac cgccacccaa
     6721 tatcttcttt ttcatctctt caaaggtata tttgccactg tagcacaatc tcgcaagttc
     6781 taatgatgga agtcctcctg ctctctcagg tgaaaatggc ccttctccat ttaaagcatc
     6841 atttacatct attacataac catgtctatg agctccaatc gaaatccctc cgcctaaatg
     6901 tgcaattata aggttggcat cctcgtattt cttctttaaa tcagaagcta agcgccttcc
     6961 aatagctttt tgatttaatg cgtgaaatat gctggatttt tggatttcag gcattcccgt
     7021 aattctggcg acatcgtcta actcatctac aacaactgga tcaactataa atgcaggtat
     7081 actatgattt ttagctaatt cgtatgcaat aatgcctcca agatttgatg catgttcacc
     7141 tcttttggcc tcttttaagt ctaataccat cctttcattt acaatgtaag tgccactttt
     7201 tatcggtttt aacaagccac ctcttcctac aatcagttag gatgttggca gacgcttgtt
     7261 gatacgaggc ttaaaattgc tgaggtaatc ggtaaattcg acccgtctat cagctgtccc
     7321 tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc
     7381 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
     7441 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
     7501 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     7561 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     7621 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     7681 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7741 ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt
     7801 ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt
     7861 tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc
     7921 cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc
     7981 cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg
     8041 aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg
     8101 tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag
     8161 agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt
     8221 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     8281 ataagttgta attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta
     8341 tcacagttaa attgctaacg cagtcaggca cctatacatg catttactta taatacagtt
     8401 ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt
     8461 tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac aataataatg
     8521 tcagatcctg tagagaccac atcatccacg gttctatact gttgacccaa tgcgtctccc
     8581 ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc atctcttcca
     8641 cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt cgcaatgtca
     8701 acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa ttctgctaac
     8761 atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa accgctaaca
     8821 atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc tattctgtat
     8881 acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt tctgtcttcg
     8941 aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt gccctccatg
     9001 gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg acctaatgct
     9061 tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca caagtttgtt
     9121 tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg agtagcagca
     9181 cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg
     9241 tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca tatgcgtata
     9301 tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga gattaccgaa
     9361 tcaaaaaaat ttcaaagaaa ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg
     9421 aattgaaaag ctagcttatc gatgggtcct tttcatcacg tgctataaaa ataattataa
     9481 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
     9541 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
     9601 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
     9661 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
     9721 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
     9781 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
     9841 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
     9901 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatctc
     9961 taagaaacca ttattatcat gacattaacc tataaaaaag gcctctcgag
//
