LOCUS       V016485                 9414 bp    DNA     circular SYN 26-JUN-2019
DEFINITION  Exported.
ACCESSION   V016485
VERSION     V016485
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9414)
  AUTHORS   Aoki SK, Lillacci G, Gupta A, Baumschlager A, Schweingruber D,
            Khammash M.
  TITLE     A universal biomolecular integral feedback controller for robust
            perfect adaptation
  JOURNAL   Nature 570, 533-537 (2019)
   PUBMED   31217585
REFERENCE   2  (bases 1 to 9414)
  AUTHORS   Aoki SK, Lillacci G.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-APR-2019) Biosystems Science and Engineering, ETH
            Zurich, Mattenstrasse 26, Basel 4058, Switzerland
REFERENCE   3  (bases 1 to 9414)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature 570,
            533-537 (2019)"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (09-APR-2019) Biosystems Science and Engineering, ETH Zurich,
            Mattenstrasse 26, Basel 4058, Switzerland"
FEATURES             Location/Qualifiers
     source          1..9414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      26..47
                     /label="UP element"
                     /note="UP element"
                     /regulatory_class="enhancer"
     regulatory      50..85
                     /label="PsigW"
                     /note="PsigW"
                     /regulatory_class="promoter"
     regulatory      112..118
                     /label="sRBS"
                     /note="sRBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             128..169
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             176..1048
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     regulatory      1139..1145
                     /label="sRBS"
                     /note="sRBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1155..1178
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             1191..1901
                     /label="superfolder GFP"
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     terminator      2206..2292
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2384..2411
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     regulatory      2494..2515
                     /label="UP element"
                     /note="UP element"
                     /regulatory_class="enhancer"
     regulatory      2518..2553
                     /label="PsigW"
                     /note="PsigW"
                     /regulatory_class="promoter"
     regulatory      2580..2586
                     /label="sRBS"
                     /note="sRBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             2596..2637
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             2644..3516
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     regulatory      3607..3613
                     /label="sRBS"
                     /note="sRBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             3623..3646
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             3659..4369
                     /label="superfolder GFP"
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     regulatory      4472..4897
                     /label="rrnB T1T2 terminator"
                     /note="rrnB T1T2 terminator"
                     /regulatory_class="terminator"
     terminator      complement(4973..5020)
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(5024..5773)
                     /gene="luxR"
                     /label="Transcriptional activator protein LuxR"
                     /note="Transcriptional activator protein LuxR from
                     Aliivibrio fischeri. Accession#: P12746"
     regulatory      complement(5802..5855)
                     /label="Pconstitutive"
                     /note="Pconstitutive"
                     /regulatory_class="promoter"
     regulatory      5906..5970
                     /label="Plux"
                     /note="Plux"
                     /regulatory_class="promoter"
     regulatory      5906..5925
                     /label="LuxR binding site"
                     /note="LuxR binding site"
                     /regulatory_class="other"
     regulatory      5971..5996
                     /label="RBS5000"
                     /note="RBS5000"
                     /regulatory_class="ribosome_binding_site"
     CDS             5997..6557
                     /gene="sigW"
                     /label="ECF RNA polymerase sigma factor SigW"
                     /note="ECF RNA polymerase sigma factor SigW from Bacillus
                     subtilis (strain 168). Accession#: Q45585"
     regulatory      6572..6658
                     /label="rrnB T1 terminator"
                     /note="rrnB T1 terminator"
                     /regulatory_class="terminator"
     regulatory      6685..6756
                     /label="PBAD"
                     /note="PBAD"
                     /regulatory_class="promoter"
     regulatory      6685..6701
                     /label="AraC I1 binding site"
                     /note="AraC I1 binding site"
                     /regulatory_class="other"
     regulatory      6706..6722
                     /label="AraC I2 binding site"
                     /note="AraC I2 binding site"
                     /regulatory_class="other"
     regulatory      6772..6797
                     /label="RBS5000"
                     /note="RBS5000"
                     /regulatory_class="ribosome_binding_site"
     CDS             6798..7061
                     /codon_start=1
                     /transl_table=11
                     /product="RsiW"
                     /label="RsiW"
                     /note="cytoplasmic RsiW domain"
                     /protein_id="QDD67922.1"
                     /translation="MSCPEQIVQLMHMHLDGDILPKDEHVLNEHLETCEKCRKHFYEME
                     KSIALVRSTSHVEAPADFTANVMAKLPKEKKRASVKRWFRTH"
     terminator      complement(7279..7322)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(7585..8173)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     terminator      complement(8261..8355)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(8388..9245)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(9246..9350)
                     /label="AmpR promoter"
ORIGIN
        1 gacgtctcta caccctgcca aaaatggaaa attttttttc aaaagtaaaa ttgaaacctt
       61 ttgaaacgaa gctcgtatac atacagagga tcctgtcaat ttccgcgata gaggaggtag
      121 atctatgggc aaaccgattc cgaacccgct gctgggcctg gatagcaccg ccgcggctga
      181 agcgcaaaat gatcccctgc tgccgggata ctcgtttaat gcccatctgg tggcgggttt
      241 aacgccgatt gaggccaacg gttatctcga tttttttatc gaccgaccgc tgggaatgaa
      301 aggttatatt ctcaatctca ccattcgcgg tcagggggtg gtgaaaaatc agggacgaga
      361 atttgtttgc cgaccgggtg atattttgct gttcccgcca ggagagattc atcactacgg
      421 tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt tactttcgtc cgcgcgccta
      481 ctggcatgaa tggcttaact ggccgtcaat atttgccaat acggggttct ttcgcccgga
      541 tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa atcattaacg ccgggcaagg
      601 ggaagggcgc tattcggagc tgctggcgat aaatctgctt gagcaattgt tactgcggcg
      661 catggaagcg attaacgagt cgctccatcc accgatggat aatcgggtac gcgaggcttg
      721 tcagtacatc agcgatcacc tggcagacag caattttgat atcgccagcg tcgcacagca
      781 tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc cagcagttag ggattagcgt
      841 cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag ctgcttttga gcaccacccg
      901 gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac gatcaactct atttctcgcg
      961 ggtatttaaa aaatgcaccg gggccagccc gagcgagttc cgtgccggtt gtgaagaaaa
     1021 agtgaatgat gtagccgtca agttgtcact cgaggcagca aataaaaatg aagagaatac
     1081 gaatgaagtt gcacaaccaa tgttaaatgc aggtcaagca aatcgccgtc gcgtttaaag
     1141 gaggtatata tatggactac aaggacgacg acgacaaggc cgccatcgag cgtaaaggcg
     1201 aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc
     1261 ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga
     1321 agttcatctg tactactggt aaactgccgg taccttggcc gactctggta acgacgctga
     1381 cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca
     1441 agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag gatgacggca
     1501 cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac cgcattgagc
     1561 tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg gaatacaatt
     1621 ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt
     1681 ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac taccagcaaa
     1741 acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg agcacgcaaa
     1801 gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg gagttcgtaa
     1861 ccgcagcggg catcacgcat ggtatggatg aactgtacaa agcggccgcg gcgaacaaaa
     1921 acgaagaaaa caccaacgaa gtgccgacct ttatgctgaa cgcgggccag gcgaaccgtc
     1981 ttcagttgta agcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     2041 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     2101 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     2161 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     2221 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2281 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     2341 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     2401 gatggccttt ttgcgtttct acaaactctg gcggtaagtc aatgatagca ctccaaagtt
     2461 tatggttcgg gaggtctaca ccctgccaaa aatggaaaat tttttttcaa aagtaaaatt
     2521 gaaacctttt gaaacgaagc tcgtatacat acagaggatc ctgtcaattt ccgcgataga
     2581 ggaggtagat ctatgggcaa accgattccg aacccgctgc tgggcctgga tagcaccgcc
     2641 gcggctgaag cgcaaaatga tcccctgctg ccgggatact cgtttaatgc ccatctggtg
     2701 gcgggtttaa cgccgattga ggccaacggt tatctcgatt tttttatcga ccgaccgctg
     2761 ggaatgaaag gttatattct caatctcacc attcgcggtc agggggtggt gaaaaatcag
     2821 ggacgagaat ttgtttgccg accgggtgat attttgctgt tcccgccagg agagattcat
     2881 cactacggtc gtcatccgga ggctcgcgaa tggtatcacc agtgggttta ctttcgtccg
     2941 cgcgcctact ggcatgaatg gcttaactgg ccgtcaatat ttgccaatac ggggttcttt
     3001 cgcccggatg aagcgcacca gccgcatttc agcgacctgt ttgggcaaat cattaacgcc
     3061 gggcaagggg aagggcgcta ttcggagctg ctggcgataa atctgcttga gcaattgtta
     3121 ctgcggcgca tggaagcgat taacgagtcg ctccatccac cgatggataa tcgggtacgc
     3181 gaggcttgtc agtacatcag cgatcacctg gcagacagca attttgatat cgccagcgtc
     3241 gcacagcatg tttgcttgtc gccgtcgcgt ctgtcacatc ttttccgcca gcagttaggg
     3301 attagcgtct taagctggcg cgaggaccaa cgtatcagcc aggcgaagct gcttttgagc
     3361 accacccgga tgcctatcgc caccgtcggt cgcaatgttg gttttgacga tcaactctat
     3421 ttctcgcggg tatttaaaaa atgcaccggg gccagcccga gcgagttccg tgccggttgt
     3481 gaagaaaaag tgaatgatgt agccgtcaag ttgtcactcg aggcagcaaa taaaaatgaa
     3541 gagaatacga atgaagttgc acaaccaatg ttaaatgcag gtcaagcaaa tcgccgtcgc
     3601 gtttaaagga ggtatatata tggactacaa ggacgacgac gacaaggccg ccatcgagcg
     3661 taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg atggtgatgt
     3721 caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta atggtaaact
     3781 gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga ctctggtaac
     3841 gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga agcagcatga
     3901 cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt cctttaagga
     3961 tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg
     4021 cattgagctg aaaggcattg actttaaaga agacggcaat atcctgggcc ataagctgga
     4081 atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa atggcattaa
     4141 agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg ctgatcacta
     4201 ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc actatctgag
     4261 cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg ttctgctgga
     4321 gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaag cggccgcggc
     4381 gaacaaaaac gaagaaaaca ccaacgaagt gccgaccttt atgctgaacg cgggccaggc
     4441 gaaccgtctt cagttgtaag catgcaagct tggctgtttt ggcggatgag agaagatttt
     4501 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     4561 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     4621 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     4681 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     4741 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     4801 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     4861 tcctgacgga tggccttttt gcgtttctac aaactctggc ggtaagtcaa tgatagcact
     4921 cgcttgataa agagcttctg cgtgacgaag caacattaca ctacggtacc tccaaaaaac
     4981 ccctcaagac ccgtttagag gccccaaggg gttatgctag ttaattttta aagtatgggc
     5041 aatcaattgc tcctgttaaa attgctttag aaatactttg gcagcggttt gttgtattga
     5101 gtttcatttg cgcattggtt aaatggaaag tgacagtacg ctcactgcaa cctaatattt
     5161 ttgaaatatc ccaagagctt tttccttcgc atgcccacgc taaacattct ttttctcttt
     5221 tggttaaatc gttgtttgat ttattatttg ctatatttat ttttcgataa ttatcaacta
     5281 gagaaggaac aattaatggt atgttcatac acgcatgtaa aaataaacta tctatatagt
     5341 tgtctttttc tgaatgtgca aaactaagca ttccgaagcc attgttagcc gtatgaatag
     5401 ggaaactaaa cccagtgata agacctgatg ttttcgcttc tttaattaca tttggagatt
     5461 ttttatttac agcattgttt tcaaatatat tccaattaat tggtgaatga ttggagttag
     5521 aataatctac tataggatca tattttatta aattagcgtc atcataatat tgcctccatt
     5581 ttttagggta attatctagg attgaaatat cagatttaac catagaatga ggataaatga
     5641 tcgcgagtaa ataatattca caatgtacca ttttagtcat atcagataag cattgattaa
     5701 tatcattatt gcttctacaa gctttaattt tattaattat tctgtatgtg tcgtcggcat
     5761 ttatgttttt catattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     5821 cgcgaaaggt tttgcgccat tcgatggcgc gccgcttcgt caggccacat agctttcttg
     5881 ttctgatcgg aacgatcgtt ggctgacctg taggatcgta caggtttacg caagaaaatg
     5941 gtttgttata gtcgaataaa tactagagaa cttctagatc gctaaaacgg gctagcatgg
     6001 aaatgatgat taaaaaaaga attaaacaag tcaaaaaagg cgaccaggat gcatttgcgg
     6061 acatcgtaga tatttacaaa gataaaattt atcagctttg ctaccgtatg cttggcaatg
     6121 tgcatgaggc ggaggatatt gcacaggaag ctttcatcag agcgtacgtt aatatcgaca
     6181 gttttgatat taaccggaaa ttttcaactt ggctttatcg aatcgcgacc aatttgacca
     6241 ttgaccgcat tcgcaaaaag aagccggatt attacctcga cgcagaggtg gctggtacgg
     6301 aaggcttgac catgtattct caaatcgtcg cagatggtgt tttgcctgaa gatgcagttg
     6361 tatcgctgga gctctcaaac acgatacagc agaaaatttt aaagcttcct gacaaataca
     6421 gaacagtcat cgtattaaag tatattgacg aactctcatt aattgaaatc ggggagattc
     6481 taaacattcc tgtggggact gtgaaaacgc ggattcacag aggcagagag gctcttagaa
     6541 aacaattaag ggatctttaa tctagaggca tcaaataaaa cgaaaggctc agtcgaaaga
     6601 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatga
     6661 cgaagcaaca ttacactagt cgactagcat ttttatccat aagattagcg gatcctacct
     6721 gacgcttttt atcgcaactc tctactgttt ctccataccc gtttttttgg aaataacgca
     6781 aagggaggcg gatcgatatg agctgtcctg aacaaattgt gcagcttatg catatgcatc
     6841 ttgatggaga tatccttcca aaagatgaac acgtattaaa tgaacatctg gagacatgcg
     6901 agaaatgcag aaagcatttt tacgagatgg agaaatccat agcgctcgta cggagcacat
     6961 cgcatgtcga agcccccgcg gattttaccg ctaatgtcat ggcaaaattg cctaaggaga
     7021 agaaaagagc ttctgtaaaa agatggttca gaacccatta agcatgcaag cttggctgtt
     7081 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     7141 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     7201 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     7261 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     7321 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     7381 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     7441 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctg
     7501 aattctctag atactacatc acagatgaca ggcaaaaggc cagcaaaagg ccaggaaccg
     7561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7621 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7681 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7741 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     8041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8221 aaactcacgt taagggattt tggtcatgac tagtgcttgg attctcacca ataaaaaacg
     8281 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     8341 ctatcaacag gagtccaagc gagctcgtaa acttggtctg acagttacca atgcttaatc
     8401 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8461 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8521 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8581 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8641 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8701 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8761 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8821 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8881 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     8941 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     9001 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     9061 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     9121 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     9181 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     9241 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     9301 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     9361 cgaaaagtgc cacctgacgg ctgtctgttc agttatgaac gaaaagtgcc acct
//
