LOCUS       V002569                 9002 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002569
VERSION     V002569
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9002)
  AUTHORS   Tian L, Lo J, Shao X, Zheng T, Olson DG, Lynd LR.
  TITLE     Identification of a ferredoxin:NAD+ oxidoreductase enzyme in
            Thermoanaerobacterium saccharolyticum and its role in ethanol
            formation
  JOURNAL   Appl. Environ. Microbiol. (2016) In press
   PUBMED   27694237
REFERENCE   2  (bases 1 to 9002)
  AUTHORS   Tian L.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAY-2016) Thayer School of Engineering, Dartmouth
            College, 14 Engineering Dr, Hanover, NH 03755, USA
REFERENCE   3  (bases 1 to 9002)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9002)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol. (2016) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (22-MAY-2016) Thayer School of Engineering, Dartmouth College, 14
            Engineering Dr, Hanover, NH 03755, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     1..1500
     CDS             1513..1530
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             2817..3551
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     misc_recomb     3609..5046
     promoter        complement(5081..5099)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5106..5122)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             5260..5559
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             5911..6702
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(6958..7815)
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7939..8527
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    8815..8836
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8851..8881
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    8889..8905
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8913..8929
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        8950..8968
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atcttttctg gcctttaatg gcgctttcgg acatgtggaa agaagttgga tggaattcaa
       61 ttatttatct ggcggcaatg actgctatag acagtgagtt atatgaagcg gcaagtattg
      121 atggagcagg taggataaga agaattttct ccataacatt accatcaata gtgccaacca
      181 taaaaatcct tttaatatta aatgcaggat ggatattaaa tgctggattt gaacaagtat
      241 ttctattgtc aaatcccatg gtcatagact attcacagac attggaattg tatgtataca
      301 attatggtat accaatgggc agattttcat ttgcgacagc agccggtata tttaactcag
      361 ttgtttcttt agcaatggtt acattagcaa atagaatatc gaaagcactt agtggtgaaa
      421 gtgcatttta atctaattta caagatttca gggggttata aagtggctaa aaataataga
      481 ataaggttga gcacagaaga tattgtattt gacacgatta attacattct tctcacgatt
      541 gtaataatag tcaccctgta tccgtttgta catgtgttgg cagtttcatt taacgatgct
      601 atagatactg tgaggggtgg aatttatcta tttccaagaa aatttacgac ttttaattac
      661 gtttcaattt taagcgatcc tgagatttac aatgctacaa tcatttcagc attaagagct
      721 gttataggtt ctgttttaaa tgtgattgcc agcataatag ttgcatatgc cataagcaga
      781 agggattttg ttttaagagg tatagtttca aaagtattta cttactcaat gtattttaca
      841 agtggcttaa ttccatatta cttgctgatt agagatttac atcttatgaa caatttcttg
      901 gtttacatct tgccagggat agtaagcgca tggaatataa tggttgttag gagttatatt
      961 gatggcttgc cagccagtct tgtggaatcg gcaaaaatag atggcgctag tgagctgagg
     1021 ataatatttt ctatcatatt tccacttagt gttccagtat tagcgacaat aacgcttttt
     1081 gttgcagttg gacaatggaa ttcgtggttt gatacaatgc ttttttgttc catgaatcca
     1141 aatttaagca cattacagta tgaattgcaa aaagtattgc agtcgactca acagtttaca
     1201 caacaagcgt cttttgatat ggggttaaga aataattcta attcagttac cccggaaagc
     1261 ataagagctg ctatgacggt agtggcagtg gttcctatta tgatggttta tccattcttg
     1321 cagaagtatt ttgtaaaggg tttaaccatt ggcagtgtta agggttagta catctttggt
     1381 tttttataaa tgctgaaagg ggggaaattt aaaaatactt attagacaaa attttttaat
     1441 tgcgaggtga tagtaaataa atttttcaat aaattaaatt tactgaggga ggaagtaaga
     1501 atgtctggtt ctcatcatca tcatcatcat ggtagatacg ttgttagaga aaatagagaa
     1561 attagcaatg gaatatacag acttagatta gaatttgatg ggattcctac gccagggcaa
     1621 tttatcatga taaattgcgg tggcaataca ttattaaaga ggccatttag catatgcgat
     1681 catgatgcag atgaaatcac aattgtttat cggataaaag gaaaaggcac ttataatctt
     1741 tcgaaggtta aagctttaga tacattagag gtaactggtc ctcatggtca tggatttgat
     1801 atgtttgata aattcaacaa tattctcatc gttggtggtg gcatagggat accaccacta
     1861 ctatttcttt caaaaaaatt gaaatcgaaa aatctaaaga ttgctcttgg ttttaaaagc
     1921 gatgtatttc ttgtggatga atttaaaaaa tatggtgatg tcgttattgc gactgaagat
     1981 gggaataccg gtattaaagg gtatgttacc gatgcaatga aagatgaaat aaaggatatt
     2041 gatatgattt atggatgtgg cccaatgcaa atgcttaaat ctgtaaaaga catggctgaa
     2101 aagtacgatg tgccttgtca aatctctatt gaagaacata tgggttgtgg aattggtgcg
     2161 tgtatggtat gtgcctgtaa aacaaaaact gattctggat ttcaatataa aaaagtctgt
     2221 aaggatggtc ctgtattttg gtcaagggag gtagtatttt gatgcaggtc gataaaccca
     2281 gcgaaccatt tgaggtgata ggtaagatta taccgaggta tgaaaacgag aattggacct
     2341 ttacagaatt actctatgaa gcgccatatt taaaaagcta ccaagacgaa gaggatgaag
     2401 aggatgagga ggcagattgc cttgaatata ttgacaatac tgataagata atatatcttt
     2461 tatatagaag atatcgccgt atgtaaggat ttcagggggc aaggcatagg cagcgcgctt
     2521 atcaatatat ctatagaatg ggcaaagcat aaaaacttgc atggactaat gcttgaaacc
     2581 caggacaata accttatagc ttgtaaattc tatcataatt gtggtttcaa aatcggctcc
     2641 gtcgatacta tgttatacgc caactttcaa aacaactttg aaaaagctgt tttctggtat
     2701 ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg tcttgttata attagcttct
     2761 tggggtatct ttaaatactg tagaaaagag gaaggaaata ataaatggcg gatcccatga
     2821 acaaaaatat aaaatattct caaaactttt taacgagtga aaaagtactc aaccaaataa
     2881 taaaacaatt gaatttaaaa gaaaccgata ccgtttacga aattggaaca ggtaaagggc
     2941 atttaacgac gaaactggct aaaataagta aacaggtaac gtctattgaa ttagacagtc
     3001 atctattcaa cttatcgtca gaaaaattaa aactgaatac tcgtgtcact ttaattcacc
     3061 aagatattct acagtttcaa ttccctaaca aacagaggta taaaattgtt gggagtattc
     3121 cttaccattt aagcacacaa attattaaaa aagtggtttt tgaaagccat gcgtctgaca
     3181 tctatctgat tgttgaagaa ggattctaca agcgtacctt ggatattcac cgaacactag
     3241 ggttgctctt gcacactcaa gtctcgattc agcaattgct taagctgcca gcggaatgct
     3301 ttcatcctaa accaaaagta aacagtgtct taataaaact tacccgccat accacagatg
     3361 ttccagataa atattggaag ctatatacgt actttgtttc aaaatgggtc aatcgagaat
     3421 atcgtcaact gtttactaaa aatcagtttc atcaagcaat gaaacacgcc aaagtaaaca
     3481 atttaagtac cgttacttat gagcaagtat tgtctatttt taatagttat ctattattta
     3541 acgggaggaa ataattctat gagtcgcttt tgtaaatttg gaaagttaca cgttactaaa
     3601 gggaattcaa aaaacaaata atctttaagt aaaaaggcag agaggaataa tcttccctct
     3661 gcctttttac tttgtgataa tttagactgt tatttctatt gatatgtcgt ttttatctaa
     3721 attgaaatga aattttctat cattaatgat taattcatat tctcctaaaa agcctgtaaa
     3781 ttcgaattca cctgtattgc ttgtgacaag acgtgtttga tgtgtccacc attcgtcttt
     3841 aataagcttt tttaattcgt aatatgatgg cttgcatgaa ttgtcctctc taagaaggcc
     3901 tgcaggcgca ccaagccatg cattttcatc gcagaaattc caccatgtga ttgattctac
     3961 cattggatgt gaaaagagag tcttataatg ttttacgact tccgttgctt gtcgttcttc
     4021 cccatcaggt gttgaaggcc aatcacttat ctggtagtca tttaggtctt ctatttcagg
     4081 tggcatcaaa tgccctgata ataatgtgtt ttctgtaaaa tgcaatggaa tattgaaatg
     4141 cgaaaatctt tcgagtactt ctaaagtttt ttcaactccc caatatcctt gatgcatgtg
     4201 tgattggatc cctatggcat caatagggat tccagcttca aggcatcctt ctatgagtat
     4261 ttcatatgaa attgatgtat taaagtcatt tataaggaga actgcttcag gattagcttt
     4321 tttagcttca ttaaaaactt cttttacgag gcgaatacgt cctaattctt tgcatatcct
     4381 ggttattcca ttgtcgtact tattaaaaat aggcattatg acaacttcat ttattacatc
     4441 ccatatattc actaatcctt taaaatcact tgcttcacgt tttatgcggg atagctgagc
     4501 ctttaatatg tcttcattgc tcatatctaa gagccaaggt gctgttacag tatgccagca
     4561 aagtggatgg ccttttacaa ggcaaccttt tgatacaagc cattctgaag cctttttgag
     4621 tctatttgtg tctggctttc ctttttcagg ctcgaacctg ccccaataaa atggaatcgt
     4681 agcatagtta aataagtcaa aaaatttctc aaaacggtct tcaatttttt ctttatcatt
     4741 ttctttcaat tcaccattgg caagaggaag agaatcaaat gctccacaac caaataaaaa
     4801 tttatgcttc acctgagata cagcgacttc tgcatctttt ataggtgaac catctttctt
     4861 aactaacttt attttggcga ttcctttgcg gtgttttaat ttatctgcag catccgcatt
     4921 cattataatc cctccaacat atgtaattgt ttataaataa aatttactct cacactttga
     4981 ataagtcaat aaatcatact tataagacta acataatatt aaggcgtttt tttgtcgcat
     5041 ttgactaagg gcgaattcgc ggccgctaaa ttcaattcgc cctatagtga gtcgtattac
     5101 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5161 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5221 gatcgccctt cccaacagtt gcgcagccta tacgtacggc agtttaaggt ttacacctat
     5281 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccg
     5341 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     5401 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     5461 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     5521 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg catgagatta
     5581 tcaaaaagga tcttcaccta gatccttttc acgtagaaag ccagtccgca gaaacggtgc
     5641 tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag
     5701 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg
     5761 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     5821 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagctct
     5881 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5941 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     6001 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     6061 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     6121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     6181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     6241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     6301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     6361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     6421 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     6481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     6541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     6601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     6661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac
     6721 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg
     6781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6841 aaatatgtat ccgctcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8041 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     8101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8221 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8281 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8341 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8401 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8461 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8521 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8581 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8641 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8701 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     8761 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     8821 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     8881 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     8941 caagctcaga attaaccctc actaaaggga ctagtcctgc aggtttaaac gaattcgccc
     9001 tt
//
