LOCUS       V002570                 9062 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002570
VERSION     V002570
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9062)
  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 9062)
  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 9062)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9062)
  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..9062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(35..53)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(60..76)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             214..513
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             865..1656
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(1912..2769)
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2893..3481
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    3769..3790
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3805..3835
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3843..3859
                     /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     3867..3883
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3904..3922
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_recomb     3957..5456
     CDS             5469..5486
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             6833..7567
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     misc_recomb     7625..9062
ORIGIN
        1 aagggcgaat tcgcggccgc taaattcaat tcgccctata gtgagtcgta ttacaattca
       61 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
      121 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
      181 ccttcccaac agttgcgcag cctatacgta cggcagttta aggtttacac ctataaaaga
      241 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga
      301 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
      361 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
      421 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
      481 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa
      541 aggatcttca cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc
      601 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
      661 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
      721 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
      781 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa
      841 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
      901 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      961 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     1021 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     1081 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     1141 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     1201 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     1261 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     1321 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     1381 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     1441 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     1501 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     1561 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     1621 cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttattaacgc ttacaatttc
     1681 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat caggtggcac
     1741 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1801 gtatccgctc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2821 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2881 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2941 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3001 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3061 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3121 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3181 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3241 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3301 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3361 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3421 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3481 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3541 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3601 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3661 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     3721 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     3781 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     3841 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     3901 cagaattaac cctcactaaa gggactagtc ctgcaggttt aaacgaattc gcccttatct
     3961 tttctggcct ttaatggcgc tttcggacat gtggaaagaa gttggatgga attcaattat
     4021 ttatctggcg gcaatgactg ctatagacag tgagttatat gaagcggcaa gtattgatgg
     4081 agcaggtagg ataagaagaa ttttctccat aacattacca tcaatagtgc caaccataaa
     4141 aatcctttta atattaaatg caggatggat attaaatgct ggatttgaac aagtatttct
     4201 attgtcaaat cccatggtca tagactattc acagacattg gaattgtatg tatacaatta
     4261 tggtatacca atgggcagat tttcatttgc gacagcagcc ggtatattta actcagttgt
     4321 ttctttagca atggttacat tagcaaatag aatatcgaaa gcacttagtg gtgaaagtgc
     4381 attttaatct aatttacaag atttcagggg gttataaagt ggctaaaaat aatagaataa
     4441 ggttgagcac agaagatatt gtatttgaca cgattaatta cattcttctc acgattgtaa
     4501 taatagtcac cctgtatccg tttgtacatg tgttggcagt ttcatttaac gatgctatag
     4561 atactgtgag gggtggaatt tatctatttc caagaaaatt tacgactttt aattacgttt
     4621 caattttaag cgatcctgag atttacaatg ctacaatcat ttcagcatta agagctgtta
     4681 taggttctgt tttaaatgtg attgccagca taatagttgc atatgccata agcagaaggg
     4741 attttgtttt aagaggtata gtttcaaaag tatttactta ctcaatgtat tttacaagtg
     4801 gcttaattcc atattacttg ctgattagag atttacatct tatgaacaat ttcttggttt
     4861 acatcttgcc agggatagta agcgcatgga atataatggt tgttaggagt tatattgatg
     4921 gcttgccagc cagtcttgtg gaatcggcaa aaatagatgg cgctagtgag ctgaggataa
     4981 tattttctat catatttcca cttagtgttc cagtattagc gacaataacg ctttttgttg
     5041 cagttggaca atggaattcg tggtttgata caatgctttt ttgttccatg aatccaaatt
     5101 taagcacatt acagtatgaa ttgcaaaaag tattgcagtc gactcaacag tttacacaac
     5161 aagcgtcttt tgatatgggg ttaagaaata attctaattc agttaccccg gaaagcataa
     5221 gagctgctat gacggtagtg gcagtggttc ctattatgat ggtttatcca ttcttgcaga
     5281 agtattttgt aaagggttta accattggca gtgttaaggg ttagtacatc tttggttttt
     5341 tataaatgct gaaagggggg aaatttaaaa atacttatta gacaaaattt tttaattgcg
     5401 aggtgatagt aaataaattt ttcaataaat taaatttact gagggaggaa gtaagaatgt
     5461 ctggttctca tcatcatcat catcatggta taaatgtgta catgccaaaa aaagtagaaa
     5521 tattgtctat aattcctcaa acagatatcg attatacttt taggctaaaa tccgatattc
     5581 tgccaagaca tggacaattt ttacaagtat ctatacctaa aatcggtgaa gccccaatat
     5641 cgataagcga ttacactgat gaatacatcg aacttaccat aagaaaagta ggcactgtaa
     5701 cagatgctat acatgaatta aaaccaggcg atttcttgtt tataagaggc ccatatggcc
     5761 acggatttcc tgtagaagat tttaaagata aaaatgtagt gatcgccgct ggcggtacag
     5821 gtctagcacc agtcaaaagc ataataaaca ggtattacag aaatcctaaa gagataaaaa
     5881 acctcaatat tttgatgggt tttaaatctc cgaaagacat actttttgaa gatgaaataa
     5941 aaaaatggaa agaaaaattt gatgttttat tgactgttga caatggagat gaaacgtgga
     6001 caggaaatac aggtttaatc acaaaattta taccagaatt aaaaatcgaa aaccctaatg
     6061 atacgatcgt aattgtcgta ggtccaccta tgatgatgaa atttacatgc cttgaatttt
     6121 taaagcgaaa tataccggat gaaaatatat gggtatcttt cgaaagaaag atgtcatgcg
     6181 gcataggaaa atgtggccat tgtaagataa atgaaaccta cgtttgctta gaaggtcctg
     6241 tattcaatta cacaaaatca aagcaacttc tcgattaatg caggtcgata aacccagcga
     6301 accatttgag gtgataggta agattatacc gaggtatgaa aacgagaatt ggacctttac
     6361 agaattactc tatgaagcgc catatttaaa aagctaccaa gacgaagagg atgaagagga
     6421 tgaggaggca gattgccttg aatatattga caatactgat aagataatat atcttttata
     6481 tagaagatat cgccgtatgt aaggatttca gggggcaagg cataggcagc gcgcttatca
     6541 atatatctat agaatgggca aagcataaaa acttgcatgg actaatgctt gaaacccagg
     6601 acaataacct tatagcttgt aaattctatc ataattgtgg tttcaaaatc ggctccgtcg
     6661 atactatgtt atacgccaac tttcaaaaca actttgaaaa agctgttttc tggtatttaa
     6721 ggttttagaa tgcaaggaac agtgaattgg agttcgtctt gttataatta gcttcttggg
     6781 gtatctttaa atactgtaga aaagaggaag gaaataataa atggcggatc ccatgaacaa
     6841 aaatataaaa tattctcaaa actttttaac gagtgaaaaa gtactcaacc aaataataaa
     6901 acaattgaat ttaaaagaaa ccgataccgt ttacgaaatt ggaacaggta aagggcattt
     6961 aacgacgaaa ctggctaaaa taagtaaaca ggtaacgtct attgaattag acagtcatct
     7021 attcaactta tcgtcagaaa aattaaaact gaatactcgt gtcactttaa ttcaccaaga
     7081 tattctacag tttcaattcc ctaacaaaca gaggtataaa attgttggga gtattcctta
     7141 ccatttaagc acacaaatta ttaaaaaagt ggtttttgaa agccatgcgt ctgacatcta
     7201 tctgattgtt gaagaaggat tctacaagcg taccttggat attcaccgaa cactagggtt
     7261 gctcttgcac actcaagtct cgattcagca attgcttaag ctgccagcgg aatgctttca
     7321 tcctaaacca aaagtaaaca gtgtcttaat aaaacttacc cgccatacca cagatgttcc
     7381 agataaatat tggaagctat atacgtactt tgtttcaaaa tgggtcaatc gagaatatcg
     7441 tcaactgttt actaaaaatc agtttcatca agcaatgaaa cacgccaaag taaacaattt
     7501 aagtaccgtt acttatgagc aagtattgtc tatttttaat agttatctat tatttaacgg
     7561 gaggaaataa ttctatgagt cgcttttgta aatttggaaa gttacacgtt actaaaggga
     7621 attcaaaaaa caaataatct ttaagtaaaa aggcagagag gaataatctt ccctctgcct
     7681 ttttactttg tgataattta gactgttatt tctattgata tgtcgttttt atctaaattg
     7741 aaatgaaatt ttctatcatt aatgattaat tcatattctc ctaaaaagcc tgtaaattcg
     7801 aattcacctg tattgcttgt gacaagacgt gtttgatgtg tccaccattc gtctttaata
     7861 agctttttta attcgtaata tgatggcttg catgaattgt cctctctaag aaggcctgca
     7921 ggcgcaccaa gccatgcatt ttcatcgcag aaattccacc atgtgattga ttctaccatt
     7981 ggatgtgaaa agagagtctt ataatgtttt acgacttccg ttgcttgtcg ttcttcccca
     8041 tcaggtgttg aaggccaatc acttatctgg tagtcattta ggtcttctat ttcaggtggc
     8101 atcaaatgcc ctgataataa tgtgttttct gtaaaatgca atggaatatt gaaatgcgaa
     8161 aatctttcga gtacttctaa agttttttca actccccaat atccttgatg catgtgtgat
     8221 tggatcccta tggcatcaat agggattcca gcttcaaggc atccttctat gagtatttca
     8281 tatgaaattg atgtattaaa gtcatttata aggagaactg cttcaggatt agctttttta
     8341 gcttcattaa aaacttcttt tacgaggcga atacgtccta attctttgca tatcctggtt
     8401 attccattgt cgtacttatt aaaaataggc attatgacaa cttcatttat tacatcccat
     8461 atattcacta atcctttaaa atcacttgct tcacgtttta tgcgggatag ctgagccttt
     8521 aatatgtctt cattgctcat atctaagagc caaggtgctg ttacagtatg ccagcaaagt
     8581 ggatggcctt ttacaaggca accttttgat acaagccatt ctgaagcctt tttgagtcta
     8641 tttgtgtctg gctttccttt ttcaggctcg aacctgcccc aataaaatgg aatcgtagca
     8701 tagttaaata agtcaaaaaa tttctcaaaa cggtcttcaa ttttttcttt atcattttct
     8761 ttcaattcac cattggcaag aggaagagaa tcaaatgctc cacaaccaaa taaaaattta
     8821 tgcttcacct gagatacagc gacttctgca tcttttatag gtgaaccatc tttcttaact
     8881 aactttattt tggcgattcc tttgcggtgt tttaatttat ctgcagcatc cgcattcatt
     8941 ataatccctc caacatatgt aattgtttat aaataaaatt tactctcaca ctttgaataa
     9001 gtcaataaat catacttata agactaacat aatattaagg cgtttttttg tcgcatttga
     9061 ct
//
