LOCUS       V002109                 8358 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002109
VERSION     V002109
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8358)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Tunable expression tools enable single-cell strain distinction in
            the gut microbiome
  JOURNAL   Cell (2017) In press
REFERENCE   2  (bases 1 to 8358)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8358)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8358)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell (2017)
            In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-MAR-2017) Microbiology "
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(29..417)
                     /direction=LEFT
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            588..697
                     /label="oriT"
                     /note="incP origin of transfer"
     regulatory      841..933
                     /regulatory_class="promoter"
     regulatory      942..964
                     /regulatory_class="ribosome_binding_site"
     CDS             965..2467
                     /codon_start=1
                     /product="Bxb1"
                     /label="Bxb1"
                     /protein_id="ARI71322.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     terminator      2487..2514
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(2746..4029)
                     /codon_start=1
                     /product="NBU integrase"
                     /label="NBU integrase"
                     /protein_id="ARI71323.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKSGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTPKYALLSTHAGRRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     misc_feature    4189..4200
                     /label="integration site"
                     /note="integration site"
     CDS             complement(4478..5335)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5336..5440)
                     /label="AmpR promoter"
     CDS             complement(5871..7793)
                     /gene="tetQ"
                     /label="Tetracycline resistance protein TetQ"
                     /note="Tetracycline resistance protein TetQ from
                     Bacteroides fragilis. Accession#: Q08425"
ORIGIN
        1 ccgacaccgc ggtggagggg aattcccatg tcagccgtta agtgttcctg tgtcactcaa
       61 aattgctttg agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac
      121 aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt
      181 aaaaccttag aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga
      241 gcttagtacg tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc
      301 atgagggttt agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg
      361 tgaaacatga gagcttagta cgtactatca acaggttgaa ctgctgatct tcagatcctc
      421 tacgccggac gcatcgtggc cggatcaatt ccgttttccg ctgcataacc ctgcttcggg
      481 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
      541 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
      601 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
      661 gtgctcaacg ggaatcctgc tctgcgaggc tggccggcta ccgccggcgt aacagatgag
      721 ggcaagcgga tggctgatga aaccaagcca accaggaagg gcagcccacc tatcacggaa
      781 ttgatccccc tcgaattgga attcgttccg tctcgattca gatctaaagg atccaatcta
      841 gagtttgcaa tggttaatct attgttaaaa tttaaagttt cacttgaact ttcaaataat
      901 gttcttatat ttgcagtgtc gaaagaaaca aaggactgat cacacaacat ttaaaaaata
      961 acaaatgaga gccctggtag tcatccgcct gtcccgcgtc accgatgcta cgacttcacc
     1021 ggagcgtcag ctggagtctt gccagcagct ctgcgcccag cgcggctggg acgtcgtcgg
     1081 ggtagcggag gatctggacg tctccggggc ggtcgatccg ttcgaccgga agcgcagacc
     1141 gaacctggcc cggtggctag cgttcgagga gcaaccgttt gacgtgatcg tggcgtaccg
     1201 ggtagatcgg ttgacccgat cgatccggca tcttcagcag ctggtccact gggccgagga
     1261 ccacaagaag ctggtcgtct ccgcgaccga agcgcacttc gatacgacga cgccgtttgc
     1321 ggcggtcgtc atcgcgctta tgggaacggt ggcgcagatg gaattagaag cgatcaaaga
     1381 gcggaaccgt tcggctgcgc atttcaatat ccgcgccggg aaataccgag gatccctgcc
     1441 gccgtgggga tacctgccta cgcgcgtgga cggggagtgg cggctggtgc cggaccctgt
     1501 gcagcgagag cgcatcctcg aggtgtatca ccgcgtcgtc gacaaccacg agccgctgca
     1561 tctggtggcc cacgacctga accggcgtgg tgtcctgtcg ccgaaggact acttcgcgca
     1621 gctgcaaggc cgcgagccgc agggccggga gtggtcggct accgcgctga agcgatcgat
     1681 gatctccgag gcgatgctcg ggtacgcgac tctgaacggt aagaccgtcc gagacgacga
     1741 cggagccccg ctggtgcggg ctgagccgat cctgacccgt gagcagctgg aggcgctgcg
     1801 cgccgagctc gtgaagacct cccgggcgaa gcccgcggtg tctaccccgt cgctgctgct
     1861 gcgggtgttg ttctgcgcgg tgtgcgggga gcccgcgtac aagttcgccg ggggaggacg
     1921 taagcacccg cgctaccgct gccgctcgat ggggttcccg aagcactgcg ggaacggcac
     1981 ggtggcgatg gccgagtggg acgcgttctg cgaggagcag gtactggatc tgctcgggga
     2041 cgcggagcgt ctggagaaag tctgggtagc gggctcggac tccgcggtcg aactcgcgga
     2101 ggtgaacgcg gagctggtgg acctgacgtc gctgatcggc tccccggcct accgggcggg
     2161 ctctccgcag cgagaagcac tggatgcccg tattgcggcg ctggccgcgc ggcaagagga
     2221 gctggagggc ctggaggctc gcccgtctgg ctgggagtgg cgcgaaaccg ggcagcggtt
     2281 cggggactgg tggcgggagc aggacaccgc ggcaaagaac acctggcttc ggtcgatgaa
     2341 cgttcggctg acgttcgacg tccgcggcgg gctgactcgc acgatcgact tcggggatct
     2401 tcaggagtac gagcagcatc tcaggctcgg cagcgtggtc gaacggctac acaccgggat
     2461 gtcgtaaatc ctagctgatt agctcgagaa ggccatcctg acggatggcc ttttttttgt
     2521 atcgagacga ctagtgtaat tgcctatctt ccagtgatgg aacagcattt gtgcattggc
     2581 tgcaacaatc agccttactt gtgcctgttc tatttccgaa ccgaccgctt gtatgaatcc
     2641 atcaaaattc gttttctcta tgttggattc cttgttgctc atattgtgat gataatttct
     2701 acaaatatag tcattggtaa ctatctatga aactgtttga tacttttata gttgattaaa
     2761 cttgttcatg gcatttgcct taatatcatc cgctatgtca atgtagggtt tcatagcttt
     2821 gtagtcgctg tgtcccgtcc atttcatgac cacctgtgcc gggattccga gagccagcgc
     2881 attgcagatg aatgtccttc ttcctgcatg ggtactgagc aaagcgtatt tgggtgtgac
     2941 ttcatcaata cgttcatttc ccttgtagta ggtttcccgt acaggctcgt tgatttctgc
     3001 cagttcgccc agctctttca ggtaatcgtt catcttctgg ttgctgatga cgggcagagc
     3061 catgtaattc tcgaaatgga tgtccttgta tttgtccagt atggctttgc tgtatttgtt
     3121 cagttcaatc gtcaggctgt cggcagtctt gactgtggtt atttcgatgt ggtcggactt
     3181 cacatcgctt cttttcagat tgcgaacatc cgaataccgc aaactcgtaa agcagcagaa
     3241 caggaaaaca tcacgcacac gttccaggta ttgcttatcc ttgggtatct ggtagtcttt
     3301 cagcttgttc agttcatccc aagtcaggaa gattactttt ttcgaggtgg ttttcagttt
     3361 cggtttgaac gtatcgtatg caatgttctg atgatgtcct ttcttgaagc tccagcgcag
     3421 gaaccatttg aggaatccca tttgcttgcc gatggtgctg tttctcatat ccttggtgtc
     3481 acgcaggaag ttgacgtatt cgttcaatcc aaactcgttg aaatagttga acgttgcatc
     3541 ctccttgaac tctttgaggt ggttcctcac tgctgcaaat ttttcatagg tggatgccgt
     3601 ccagttattc tggttaccgc actcttttac aaactcatcg aacacctccc aaaagctgac
     3661 aggggcttct tccggctgtt cttcgctggt gtctttcatt ctcatgttga aagcttcctt
     3721 caactgttgg gtcgttggca tgacctcctg cacctcaaat tccttgaaaa tattctggat
     3781 ttcggcatag tatttcagca agtccgtatt gatttcggct gcactttgct ttagcttgtt
     3841 ggtacatccg ctctttaccc gctgcttatc tgcatcccat ttggctacgt caatccggta
     3901 gcccgttgta aactcgatgc gttggctggc aaagatgaca cgcatacgga tgggtacgtt
     3961 ctctacgatt ggcacaccgt tctttttccg gctctccaat gcaaaaatga tgttgcgctt
     4021 gatattcata attgggtgcg tttgaaattc tacacccaaa tatacaccca attattgaga
     4081 tagcaaaaga catttagaaa catttacttt tactctatat tgtaatttac acttgattat
     4141 cagtcgtttg cagtcttatg atattctgtg aaagtataag ttcgagagcc tgtctctccg
     4201 caaaaaacgc tgaaaatcag cagattgcaa aacaaacacc ctgttttaca cccaagaatg
     4261 taaagtcggc tgtttttgtt ttatttaaga taatacaacc actacataat aaaagagtag
     4321 cgatattaaa agaatccgat gagaaaagac taatatttat ctatccattc agtttgattt
     4381 ttcaggactt tacatcgtcc tgaaagtatt tgttggtacc ggtaccgagg acgcgacagc
     4441 tgcagacgcg ttatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4501 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4561 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc
     4621 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4681 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4741 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4801 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4861 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4921 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4981 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5041 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5101 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5161 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5221 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5281 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5341 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5401 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5461 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5521 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5581 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5641 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     5701 ttgtactgag agtgcaccat agagctcgtc tattttttta ttgccaagtt ctaatgcttc
     5761 tatctcctgc cattcataga ggctattata tgccatacat ccgataagaa tggagataag
     5821 tatatatccc aactgtattg ccttatagaa atttcctgac cgctccatta ttttgatgac
     5881 attgattttt ggaacatgaa taaaagttta tctttttcgt tcatgcggat attatcagaa
     5941 taaccgcctt ttgttatttg atacccgcat ggcttaacca taaaaatgcc taagccctta
     6001 gtgtatgaac ttacttctga tgcatagtct ttacttgtat ttaatggaac tttcccttta
     6061 atatgacacc actcattatt gcaactgatg tcttcaatct cagacatcat tttttgcaaa
     6121 tctgtaatag ctttggaact tgccgcttgg ggtatctgca actcaaaata gagcatcggt
     6181 tcgagaatgt ccacacctga ctgttgcaag gccagcctga agacataagg ggtcagctgt
     6241 ctgaaatcag caggtgtact taccgggcta taatactcgg cttgagtaaa agttactttc
     6301 agatcagtca cttcccatcc atgtaacccg gattggcaag acatacgaat cccttcaaaa
     6361 acggcatttt gaaaagaatg gttcagataa ccataggaga tgtcactttc gatttgcaac
     6421 cctgtcccta acggtaaggg ttcaagagtc agccctattg tggcccaata agggttgggc
     6481 ggcacttcga tctgaataat cttattgacc ttttttacag gtcgttcttt gtatatagtc
     6541 ttgatctcat caaaatggac ctttacggaa aatcgttctt ccagcaatgt ctgtatgatt
     6601 tccttttggg ttaaaccata taacgagatt tccaattcat cactatatga gtttatggaa
     6661 aaggacaaag acgggtcttc aatccacaat gtattcagag cggatatcac cttgcttctc
     6721 tcttcgggcc tgtctggccg gacggaggat ttgagagcgg gatgctgatg cgataatcct
     6781 tgaatcaaac aaggttcagc acctaaataa tttccgattc gaaaatcatc catatcctct
     6841 acaatcgcga tatcattggc gcccacttca tcaacattta tctctctgcc ctgattgata
     6901 gtttttagat ttttaatctt gatgaatttt tccgaatcgt tgattcttac aacgtctcga
     6961 agtctcagac ttccgtcaat tatttttaga aaacttcttt tatgtccttt ggggtcatgc
     7021 tctatcttat aaagataaga tgaaagtctg tttgaaaccg atgccggagg aagtataaaa
     7081 gaagtgatgg cgtccaacaa ctcattgata ccgatattga acattgctga tccatgtagc
     7141 accggataga ctttggcttt tgccacaaga gcgattatcg tattccaata atcagccggt
     7201 gaaatttcgc tatccgccaa atatcgttct aatatattgt cgtcatggtt gcatacaaat
     7261 tctttgtatt cttcctttat atatgtttgg gagcaaaccg gataaaccga tccatcgaca
     7321 acattttgca taaacaggac atcttgagac agatttgctt ttatatccag atacaaacgc
     7381 tccaaattca caccggctcg gtcaatctta ttgataaata taattgtcgg gatttgcagc
     7441 ttctgtaaag tattgaacag caactttgtc tgcgcttgta tgccttcctt tgcggataag
     7501 atgaggactg ctccatcaag cattttgaat gtccgctcca cttccgcaat aaaatccatg
     7561 tgtcccggag tgtcaatgat attgcatttc acaccattcc agataataga tgtcgtagaa
     7621 gcccgaacag taattcctct acgtttctct atatccatag agtccgttat ggtgtcacca
     7681 ttatccacac agccgcactt ttccgttgct ccactggcaa acagcagatt ctcggttacg
     7741 gaagtttttc ctgcatcaat gtgagcaaga attcctaaat ttataatatt catttggatt
     7801 aagcaataat atactacaat agatgcattg tcgaaacgca ccttttaata cctcctcgta
     7861 gcatatgaga actacaggat tcctaactcg tattaatatg tatattatta ctgccgcata
     7921 acgattacaa aattacacaa aaaaatatat ctaacaaaag taggaggatt ttttaacttt
     7981 ttaccataga cattttatta gggaaacgta ggttcaactg gtaagtacaa ataagtagaa
     8041 atgtttgaac aacactgttt taggaagttg aaaaatcaag tttctctctc ggtatagcgt
     8101 ttattttggc caatatcttc tttagtttgg catttgagta tttcccattc gtgttctatt
     8161 tagctcctta ttatgagtat gtagcaagtt gcttatcaaa ttcagcctct ttgagggtca
     8221 aatatggttt tgcaaatggt gactgacaag acataaacaa ggtcagcagg attttttaca
     8281 taaatggaca tgaatgtata taacggggtt tgagggccaa tggaacgaaa acgtacgtta
     8341 aggagataac gcgtggta
//
