LOCUS       62056_19905        4403 bp DNA     circular SYN 26-JUN-2019
DEFINITION  Cloning vector pSKA417, complete sequence.
ACCESSION   MK775703
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4403)
  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 4403)
  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 4403)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature 570,
            533-537 (2019)"
COMMENT     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..4403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..25
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      26..31
                     /regulatory_class="minus_35_signal"
     regulatory      32..50
                     /label=TetR tetO2 binding site
                     /note="TetR tetO2 binding site"
                     /regulatory_class="other"
     regulatory      49..54
                     /regulatory_class="minus_10_signal"
     regulatory      67..73
                     /label=sRBS
                     /note="sRBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             80..2443
                     /codon_start=1
                     /transl_table=11
                     /product="mfLon"
                     /label=mfLon
                     /protein_id="QDD67903.1"
                     /translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
                     QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
                     DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
                     DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
                     QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
                     RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
                     KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
                     DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
                     PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
                     DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
                     EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
                     NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPAAG
                     ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
                     ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTK"
     terminator      2455..2541
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      complement(2711..3256)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(3370..3464)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3479..4267)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
ORIGIN
        1 gacgtctccc tatcagtgat agagattgac atccctatca gtgatagaga taatgtgtgg
       61 tcacacagga ggtagatcta tgagtaaaaa aataaaactg cctattttcc aaatacgcgg
      121 ttcatttatt gttcctggaa ttaaagagaa tttagaagtt ggaagaaaaa acacattagc
      181 gagcgttaat tatgcgatta aaaattcaaa taatcaaatg atcgcaatac cgcagattga
      241 tgcatctgtt gaaaaacctg aattttcaga tttacatgaa tttggaattt taattgattt
      301 tgaagtaatc aaagaatgga aagataattc attaactatt agcacaaacc ctattcaacg
      361 atgtaaagta attagctttt ttgaaaatga agatcaagtg ccgtatgcgg aagttgaatt
      421 aattgaatca attaatgatt ttagtgatga agagttaaaa gaattaattg aaaaaatctc
      481 tgatgctatc aaaactaaag ctagtttagt aacaaaacaa attaaacagt taatatcagg
      541 tgaatctgat gatttgagct tagcttttga ttcaattatg tttaaactgg ctccgtcaaa
      601 aatattgaca aatccggaat acataacatc tccgtcatta aaaacacgat ggtcaattat
      661 tgaaaaaata atttttgctg aagatggaat cataacaaga aatgcagaat caattgatgc
      721 agcacgacaa aagaatgaaa ttgaacaaga actgaaccat aaattaaaag aaaaaatgga
      781 taaacaacaa aaagaatatt atctgcgaga aaaaatgcga attatcaaag atgaattaga
      841 agatgaagat gattctgatg acagttcatt agaaaaatat aaagaacgtt tagctaaaga
      901 acctttccct gaagaagtca aacgaaaaat tatggcttct atcaaacgag ttgaagcact
      961 tcaatcagga actcctgaat ggaatactga gaaaaattac attgattgga tgatgagtat
     1021 tccctggtgg gaagaaactg aagacttaac tgatttaaaa tatgctaaaa aaattttaga
     1081 caaacatcat tatggtatga aaaaagttaa agaaagaatt attgagtact tagctgttaa
     1141 aactaaaaca aaatcactaa aagcaccaat tattacatta gttggtcccc caggtgtagg
     1201 taaaacaagt ttagcaaaat caattgcaga agcagttgga aaaaactttg ttaaagtaag
     1261 tttaggtgga gttaaagatg aatcagaaat tcgtggacat agaaaaactt atgttggttc
     1321 aatgccagga agaattattc aaacaatgaa aagagcaaaa gtaaaaaacc cattattctt
     1381 gcttgatgaa attgataaaa tggcatctga tcacagaggt gatccagcaa gtgcaatgtt
     1441 agaggttctt gacccagaac aaaacaaaga gttttcagat cattatattg aagaaccata
     1501 tgatctaagc caagttatgt ttatagcaac tgctaactat cctgaagata ttcctgaagc
     1561 cttatatgac cgtatggaaa tcataaattt atctagctat actgaaattg aaaaagttaa
     1621 gatagcacaa gattatttag ttccaaaagc aatagaacaa catgaattaa cttcagaaga
     1681 aatatcattt actgaaggtg caattaatga aattattaaa tattacacta gagaagctgg
     1741 tgtgcgtcaa ttagaaagac atattaattc tattattaga aagtatattg ttaaaaatct
     1801 aaatggtgaa atggataaaa ttgttattga tgaaaaacaa gtaaatgatt tattaggtaa
     1861 aagaattttt gatcacactg aaaaacaaga agaatctcaa attggtgtgg ttacaggttt
     1921 agcatataca caatttggtg gagatatttt accaattgaa gtaagtttat accctggtaa
     1981 aggaaattta attcttacag gtaaacttgg tgaagttatg aaagaatctg caacaattgc
     2041 gttaacttat gttaaatcaa actttgaaaa atttggagta gataaaaaag tatttgaaga
     2101 aaatgatatt catgttcacg ttcctgaagg agctgttcca aaagatggac cagctgctgg
     2161 tatcacaatt acaactgctt taatttctgc actttcagat aaaccagttt caaaagaaat
     2221 tggaatgact ggtgaaatca cactaagagg aaatgtattg ccaattggtg gattaagaga
     2281 aaaatcaatt tctgcttcaa gaagtggatt gaaaacaatc attattccaa agaaaaatga
     2341 aagagactta gacgaaattc cagatgaagt aaaagcaaaa ctaaaaatta tacctgctga
     2401 aaaatatgaa gaggtatttg caatagtttt taaaacaaaa taatctagag gcatcaaata
     2461 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2521 gctctcctga gtaggacaaa tccgccgccc tagacctagg ggatatattc cgcttcctcg
     2581 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     2641 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     2701 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     2761 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     2821 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     2881 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     2941 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     3001 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     3061 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     3121 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     3181 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     3241 accaaaacga tctcaagaag atcatcttat taatcagata aaatattact agatttcagt
     3301 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgttact
     3361 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     3421 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcttatt
     3481 tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg
     3541 cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga
     3601 cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg
     3661 cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct
     3721 catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa
     3781 atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa
     3841 cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct
     3901 cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag
     3961 ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga
     4021 gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc
     4081 gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct
     4141 tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat
     4201 ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt
     4261 ccctcatgcg aaacgatcct catcctgtct cttgatcaga tattgatccc ctgcgccatc
     4321 agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag
     4381 agggcgcccc agctggcaat tcc
//
