LOCUS       V002108                 8343 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002108
VERSION     V002108
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8343)
  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 8343)
  AUTHORS   Whitaker WR, Shepherd ES, Sonnenburg JL.
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8343)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8343)
  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..8343
                     /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..2452
                     /codon_start=1
                     /product="TP901"
                     /label="TP901"
                     /protein_id="ARI71326.1"
                     /translation="MTKKVAIYTRVSTTNQAEEGFSIDEQIDRLTKYAEAMGWQVSDTY
                     TDAGFSGAKLERPAMQRLINDIENKAFDTVLVYKLDRLSRSVRDTLYLVKDVFTKNKID
                     FISLNESIDTSSAMGSLFLTILSAINEFERENIKERMTMGKLGRAKSGKSMMWTKTAFG
                     YYHNRKTGILEIVPLQATIVEQIFTDYLSGISLTKLRDKLNESGHIGKDIPWSYRTLRQ
                     TLDNPVYCGYIKFKDSLFEGMHKPIIPYETYLKVQKELEERQQQTYERNNNPRPFQAKY
                     MLSGMARCGYCGAPLKIVLGHKRKDGSRTMKYHCANRFPRKTKGITVYNDNKKCDSGTY
                     DLSNLENTVIDNLIGFQENNDSLLKIINGNNQPILDTSSFKKQISQIDKKIQKNSDLYL
                     NDFITMDELKDRTDSLQAEKKLLKAKISENKFNDSTDVFELVKTQLGSIPINELSYDNK
                     KKIVNNLVSKVDVTADNVDIIFKFQLATGAAKDENYA"
     terminator      2472..2499
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(2731..4014)
                     /codon_start=1
                     /product="NBU integrase"
                     /label="NBU integrase"
                     /protein_id="ARI71327.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKSGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTPKYALLSTHAGRRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     misc_feature    4174..4185
                     /label="integration site"
                     /note="integration site"
     CDS             complement(4463..5320)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(5321..5425)
                     /label="AmpR promoter"
     CDS             complement(5856..7778)
                     /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 acaaatgact aagaaagtag caatctatac acgagtatcc actactaacc aagcagagga
     1021 aggcttctca attgatgagc aaattgaccg tttaacaaaa tatgctgaag caatggggtg
     1081 gcaagtatct gatacttata ctgatgctgg tttttcaggg gccaaacttg aacgcccagc
     1141 aatgcaaaga ttaatcaacg atatcgagaa taaagctttt gatacagttc ttgtatataa
     1201 gctagaccgc ctttcacgta gtgtaagaga tactctttat cttgttaagg atgtgttcac
     1261 aaaaaataaa atagacttta tctcgcttaa tgaaagtatt gatacttctt ctgctatggg
     1321 tagcttgttt ctcactattc tttctgcaat taatgagttt gaaagagaga atataaaaga
     1381 acgcatgact atgggtaaac tagggcgagc gaaatctggt aagtctatga tgtggactaa
     1441 gacagctttt gggtattacc acaacagaaa gacaggtata ttagaaattg ttcctttaca
     1501 agctacaata gttgaacaaa tattcactga ttatttatca ggaatatcac ttacaaaatt
     1561 aagagataaa ctcaatgaat ctggacacat cggtaaagat ataccgtggt cttatcgtac
     1621 cctaagacaa acacttgata atccagttta ctgtggttat atcaaattta aggacagcct
     1681 atttgaaggt atgcacaaac caattatccc ttatgagact tatttaaaag ttcaaaaaga
     1741 gctagaagaa agacaacagc agacttatga aagaaataac aaccctagac ctttccaagc
     1801 taaatatatg ctgtcaggga tggcaaggtg cggttactgt ggagcacctt taaaaattgt
     1861 tcttggccac aaaagaaaag atggaagccg cactatgaaa tatcactgtg caaatagatt
     1921 tcctcgaaaa acaaaaggaa ttacagtata taatgacaat aaaaagtgtg attcaggaac
     1981 ttatgattta agtaatttag aaaatactgt tattgacaac ctgattggat ttcaagaaaa
     2041 taatgactcc ttattgaaaa ttatcaatgg caacaaccaa cctattcttg atacttcgtc
     2101 atttaaaaag caaatttcac agatcgataa aaaaatacaa aagaactctg atttgtacct
     2161 aaatgatttt atcactatgg atgagttgaa agatcgtact gattcccttc aggctgagaa
     2221 aaagctgctt aaagctaaga ttagcgaaaa taaatttaat gactctactg atgtttttga
     2281 gttagttaaa actcagttgg gctcaattcc gattaatgaa ctatcatatg ataataaaaa
     2341 gaaaatcgtc aacaaccttg tatcaaaggt tgatgttact gctgataatg tagatatcat
     2401 atttaaattc caactcgcta ccggtgctgc taaggacgaa aactacgctt aaatcctagc
     2461 tgattagctc gagaaggcca tcctgacgga tggccttttt tttgtatcga gacgactagt
     2521 gtaattgcct atcttccagt gatggaacag catttgtgca ttggctgcaa caatcagcct
     2581 tacttgtgcc tgttctattt ccgaaccgac cgcttgtatg aatccatcaa aattcgtttt
     2641 ctctatgttg gattccttgt tgctcatatt gtgatgataa tttctacaaa tatagtcatt
     2701 ggtaactatc tatgaaactg tttgatactt ttatagttga ttaaacttgt tcatggcatt
     2761 tgccttaata tcatccgcta tgtcaatgta gggtttcata gctttgtagt cgctgtgtcc
     2821 cgtccatttc atgaccacct gtgccgggat tccgagagcc agcgcattgc agatgaatgt
     2881 ccttcttcct gcatgggtac tgagcaaagc gtatttgggt gtgacttcat caatacgttc
     2941 atttcccttg tagtaggttt cccgtacagg ctcgttgatt tctgccagtt cgcccagctc
     3001 tttcaggtaa tcgttcatct tctggttgct gatgacgggc agagccatgt aattctcgaa
     3061 atggatgtcc ttgtatttgt ccagtatggc tttgctgtat ttgttcagtt caatcgtcag
     3121 gctgtcggca gtcttgactg tggttatttc gatgtggtcg gacttcacat cgcttctttt
     3181 cagattgcga acatccgaat accgcaaact cgtaaagcag cagaacagga aaacatcacg
     3241 cacacgttcc aggtattgct tatccttggg tatctggtag tctttcagct tgttcagttc
     3301 atcccaagtc aggaagatta cttttttcga ggtggttttc agtttcggtt tgaacgtatc
     3361 gtatgcaatg ttctgatgat gtcctttctt gaagctccag cgcaggaacc atttgaggaa
     3421 tcccatttgc ttgccgatgg tgctgtttct catatccttg gtgtcacgca ggaagttgac
     3481 gtattcgttc aatccaaact cgttgaaata gttgaacgtt gcatcctcct tgaactcttt
     3541 gaggtggttc ctcactgctg caaatttttc ataggtggat gccgtccagt tattctggtt
     3601 accgcactct tttacaaact catcgaacac ctcccaaaag ctgacagggg cttcttccgg
     3661 ctgttcttcg ctggtgtctt tcattctcat gttgaaagct tccttcaact gttgggtcgt
     3721 tggcatgacc tcctgcacct caaattcctt gaaaatattc tggatttcgg catagtattt
     3781 cagcaagtcc gtattgattt cggctgcact ttgctttagc ttgttggtac atccgctctt
     3841 tacccgctgc ttatctgcat cccatttggc tacgtcaatc cggtagcccg ttgtaaactc
     3901 gatgcgttgg ctggcaaaga tgacacgcat acggatgggt acgttctcta cgattggcac
     3961 accgttcttt ttccggctct ccaatgcaaa aatgatgttg cgcttgatat tcataattgg
     4021 gtgcgtttga aattctacac ccaaatatac acccaattat tgagatagca aaagacattt
     4081 agaaacattt acttttactc tatattgtaa tttacacttg attatcagtc gtttgcagtc
     4141 ttatgatatt ctgtgaaagt ataagttcga gagcctgtct ctccgcaaaa aacgctgaaa
     4201 atcagcagat tgcaaaacaa acaccctgtt ttacacccaa gaatgtaaag tcggctgttt
     4261 ttgttttatt taagataata caaccactac ataataaaag agtagcgata ttaaaagaat
     4321 ccgatgagaa aagactaata tttatctatc cattcagttt gatttttcag gactttacat
     4381 cgtcctgaaa gtatttgttg gtaccggtac cgaggacgcg acagctgcag acgcgttatg
     4441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4561 agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc
     4621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5161 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5221 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5281 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5341 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5401 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     5461 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5521 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5581 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5641 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     5701 accatagagc tcgtctattt ttttattgcc aagttctaat gcttctatct cctgccattc
     5761 atagaggcta ttatatgcca tacatccgat aagaatggag ataagtatat atcccaactg
     5821 tattgcctta tagaaatttc ctgaccgctc cattattttg atgacattga tttttggaac
     5881 atgaataaaa gtttatcttt ttcgttcatg cggatattat cagaataacc gccttttgtt
     5941 atttgatacc cgcatggctt aaccataaaa atgcctaagc ccttagtgta tgaacttact
     6001 tctgatgcat agtctttact tgtatttaat ggaactttcc ctttaatatg acaccactca
     6061 ttattgcaac tgatgtcttc aatctcagac atcatttttt gcaaatctgt aatagctttg
     6121 gaacttgccg cttggggtat ctgcaactca aaatagagca tcggttcgag aatgtccaca
     6181 cctgactgtt gcaaggccag cctgaagaca taaggggtca gctgtctgaa atcagcaggt
     6241 gtacttaccg ggctataata ctcggcttga gtaaaagtta ctttcagatc agtcacttcc
     6301 catccatgta acccggattg gcaagacata cgaatccctt caaaaacggc attttgaaaa
     6361 gaatggttca gataaccata ggagatgtca ctttcgattt gcaaccctgt ccctaacggt
     6421 aagggttcaa gagtcagccc tattgtggcc caataagggt tgggcggcac ttcgatctga
     6481 ataatcttat tgaccttttt tacaggtcgt tctttgtata tagtcttgat ctcatcaaaa
     6541 tggaccttta cggaaaatcg ttcttccagc aatgtctgta tgatttcctt ttgggttaaa
     6601 ccatataacg agatttccaa ttcatcacta tatgagttta tggaaaagga caaagacggg
     6661 tcttcaatcc acaatgtatt cagagcggat atcaccttgc ttctctcttc gggcctgtct
     6721 ggccggacgg aggatttgag agcgggatgc tgatgcgata atccttgaat caaacaaggt
     6781 tcagcaccta aataatttcc gattcgaaaa tcatccatat cctctacaat cgcgatatca
     6841 ttggcgccca cttcatcaac atttatctct ctgccctgat tgatagtttt tagattttta
     6901 atcttgatga atttttccga atcgttgatt cttacaacgt ctcgaagtct cagacttccg
     6961 tcaattattt ttagaaaact tcttttatgt cctttggggt catgctctat cttataaaga
     7021 taagatgaaa gtctgtttga aaccgatgcc ggaggaagta taaaagaagt gatggcgtcc
     7081 aacaactcat tgataccgat attgaacatt gctgatccat gtagcaccgg atagactttg
     7141 gcttttgcca caagagcgat tatcgtattc caataatcag ccggtgaaat ttcgctatcc
     7201 gccaaatatc gttctaatat attgtcgtca tggttgcata caaattcttt gtattcttcc
     7261 tttatatatg tttgggagca aaccggataa accgatccat cgacaacatt ttgcataaac
     7321 aggacatctt gagacagatt tgcttttata tccagataca aacgctccaa attcacaccg
     7381 gctcggtcaa tcttattgat aaatataatt gtcgggattt gcagcttctg taaagtattg
     7441 aacagcaact ttgtctgcgc ttgtatgcct tcctttgcgg ataagatgag gactgctcca
     7501 tcaagcattt tgaatgtccg ctccacttcc gcaataaaat ccatgtgtcc cggagtgtca
     7561 atgatattgc atttcacacc attccagata atagatgtcg tagaagcccg aacagtaatt
     7621 cctctacgtt tctctatatc catagagtcc gttatggtgt caccattatc cacacagccg
     7681 cacttttccg ttgctccact ggcaaacagc agattctcgg ttacggaagt ttttcctgca
     7741 tcaatgtgag caagaattcc taaatttata atattcattt ggattaagca ataatatact
     7801 acaatagatg cattgtcgaa acgcaccttt taatacctcc tcgtagcata tgagaactac
     7861 aggattccta actcgtatta atatgtatat tattactgcc gcataacgat tacaaaatta
     7921 cacaaaaaaa tatatctaac aaaagtagga ggatttttta actttttacc atagacattt
     7981 tattagggaa acgtaggttc aactggtaag tacaaataag tagaaatgtt tgaacaacac
     8041 tgttttagga agttgaaaaa tcaagtttct ctctcggtat agcgtttatt ttggccaata
     8101 tcttctttag tttggcattt gagtatttcc cattcgtgtt ctatttagct ccttattatg
     8161 agtatgtagc aagttgctta tcaaattcag cctctttgag ggtcaaatat ggttttgcaa
     8221 atggtgactg acaagacata aacaaggtca gcaggatttt ttacataaat ggacatgaat
     8281 gtatataacg gggtttgagg gccaatggaa cgaaaacgta cgttaaggag ataacgcgtg
     8341 gta
//
