LOCUS       sequence-178102-        6999 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic gene circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  AUTHORS   Rubens JR, Selvaggio G, Lu TK
  TITLE     Synthetic mixed-signal computation in living cells.
  JOURNAL   Nat Commun. 2016 Jun 3;7:11658. doi: 10.1038/ncomms11658.
  PUBMED    27255669
REFERENCE   2  (bases 1 to 6999)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2016-06-3"; volume: "7:11658. doi"; pages: " 
            10.1038/ncomms11658"
COMMENT     Sequence Label: pZS1katGp-RBS31-PhiC31-proD-oxyR
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             379..2193
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     terminator      2301..2387
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3530..3616
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4161..5111)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      5159..5381
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5879..5973)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(6003..6794,6795..6863))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6626..6645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6864..6968)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtggctttt atgaaaatca cacagtgatc acaaatttta aacagagcac aaaatgctgc
       61 ctcgaaatga gggcgggaaa ataaggttat cagccttgtt ttctccctca ttacttgaag
      121 gatatgaagc taaaaccctt ttttataaag catttgtccg aattcggaca taatcaaaaa
      181 agcttaatta agatcaattt gatctacatc tctttaacca acaatatgta agatctcaac
      241 tatcgcatcc gtggattaat tcaattataa cttctctcta acgctgtgta tcgtaacggt
      301 aacactgtag aggggagcac attgatgcga attctcacac aggaaaccgg taccatgaca
      361 caaggggttg tgaccggggt ggacacgtac gcgggtgctt acgaccgtca gtcgcgcgag
      421 cgcgaaaatt cgagcgcagc aagcccagcg acacagcgta gcgccaacga agacaaggcg
      481 gccgaccttc agcgcgaagt cgagcgcgac gggggccggt tcaggttcgt cgggcatttc
      541 agcgaagcgc cgggcacgtc ggcgttcggg acggcggagc gcccggagtt cgaacgcatc
      601 ctgaacgaat gccgcgccgg gcggctcaac atgatcattg tctatgacgt gtcgcgcttc
      661 tcgcgcctga aggtcatgga cgcgattccg attgtctcgg aattgctcgc cctgggcgtg
      721 acgattgttt ccactcagga aggcgtcttc cggcagggaa acgtcatgga cctgattcac
      781 ctgattatgc ggctcgacgc gtcgcacaaa gaatcttcgc tgaagtcggc gaagattctc
      841 gacacgaaga accttcagcg cgaattgggc gggtacgtcg gcgggaaggc gccttacggc
      901 ttcgagcttg tttcggagac gaaggagatc acgcgcaacg gccgaatggt caatgtcgtc
      961 atcaacaagc ttgcgcactc gaccactccc cttaccggac ccttcgagtt cgagcccgac
     1021 gtaatccggt ggtggtggcg tgagatcaag acgcacaaac accttccctt caagccgggc
     1081 agtcaagccg ccattcaccc gggcagcatc acggggcttt gtaagcgcat ggacgctgac
     1141 gccgtgccga cccggggcga gacgattggg aagaagaccg cttcaagcgc ctgggacccg
     1201 gcaaccgtta tgcgaatcct tcgggacccg cgtattgcgg gcttcgccgc tgaggtgatc
     1261 tacaagaaga agccggacgg cacgccgacc acgaagattg agggttaccg cattcagcgc
     1321 gacccgatca cgctccggcc ggtcgagctt gattgcggac cgatcatcga gcccgctgag
     1381 tggtatgagc ttcaggcgtg gttggacggc agggggcgcg gcaaggggct ttcccggggg
     1441 caagccattc tgtccgccat ggacaagctg tactgcgagt gtggcgccgt catgacttcg
     1501 aagcgcgggg aagaatcgat caaggactct taccgctgcc gtcgccggaa ggtggtcgac
     1561 ccgtccgcac ctgggcagca cgaaggcacg tgcaacgtca gcatggcggc actcgacaag
     1621 ttcgttgcgg aacgcatctt caacaagatc aggcacgccg aaggcgacga agagacgttg
     1681 gcgcttctgt gggaagccgc ccgacgcttc ggcaagctca ctgaggcgcc tgagaagagc
     1741 ggcgaacggg cgaaccttgt tgcggagcgc gccgacgccc tgaacgccct tgaagagctg
     1801 tacgaagacc gcgcggcagg cgcgtacgac ggacccgttg gcaggaagca cttccggaag
     1861 caacaggcag cgctgacgct ccggcagcaa ggggcggaag agcggcttgc cgaacttgaa
     1921 gccgccgaag ccccgaagct tccccttgac caatggttcc ccgaagacgc cgacgctgac
     1981 ccgaccggcc ctaagtcgtg gtgggggcgc gcgtcagtag acgacaagcg cgtgttcgtc
     2041 gggctcttcg tagacaagat cgttgtcacg aagtcgacta cgggcagggg gcagggaacg
     2101 cccatcgaga agcgcgcttc gatcacgtgg gcgaagccgc cgaccgacga cgacgaagac
     2161 gacgcccagg acggcacgga agacgtagcg gcgaggcctg cagcaaacga cgaaaactac
     2221 gctgcagcag tttagacaca tggcatggat gaactataca aataacccgg gggatcccat
     2281 ggtacgcgtg ctagaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     2341 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgccctaga
     2401 cctagcacag ctaacaccac gtcgtcccta tctgctgccc taggtctatg agtggttgct
     2461 ggataacttt acgggcatgc ataaggctcg tataatatat tcagggagac cacaacggtt
     2521 tccctctaca aataattttg tttaactttg aattcttcac acaggaaacc ggtaccatga
     2581 atattcgtga tcttgagtac ctggtggcat tggctgaaca ccgccatttt cggcgtgcgg
     2641 cagattcctg ccacgttagc cagccgacgc ttagcgggca aattcgtaag ctggaagatg
     2701 agctgggcgt gatgttgctg gagcggacca gccgtaaagt gttgttcacc caggcgggaa
     2761 tgctgctggt ggatcaggcg cgtaccgtgc tgcgtgaggt gaaagtcctt aaagagatgg
     2821 caagccagca gggcgagacg atgtccggac cgctgcacat tggtttgatt cccacagttg
     2881 gaccgtacct gctaccgcat attatcccta tgctgcacca gacctttcca aagctggaaa
     2941 tgtatctgca tgaagcacag acccaccagt tactggcgca actggacagc ggcaaactcg
     3001 attgcgtgat cctcgcgctg gtgaaagaga gcgaagcatt cattgaagtg ccgttgtttg
     3061 atgagccaat gttgctggct atctatgaag atcacccgtg ggcgaaccgc gaatgcgtac
     3121 cgatggccga tctggcaggg gaaaaactgc tgatgctgga agatggtcac tgtttgcgcg
     3181 atcaggcaat gggtttctgt tttgaagccg gggcggatga agatacacac ttccgcgcga
     3241 ccagcctgga aactctgcgc aacatggtgg cggcaggtag cgggatcact ttactgccag
     3301 cgctggctgt gccgccggag cgcaaacgcg atggggttgt ttatctgccg tgcattaagc
     3361 cggaaccacg ccgcactatt ggcctggttt atcgtcctgg ctcaccgctg cgcagccgct
     3421 atgagcagct ggcagaggcc atccgcgcaa gaatggatgg ccatttcgat aaagttttaa
     3481 aacaggcggt ttaacccggg ggatcccatg gtacgcgtgc tagaggcatc aaataaaacg
     3541 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     3601 cctgagtagg acaaatccgc cgccctagac ctagggccta gggtacgggt tttgctgccc
     3661 gcaaacgggc tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc
     3721 ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag
     3781 gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca
     3841 ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt
     3901 ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg
     3961 ttacattgtc gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct
     4021 ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg
     4081 atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag
     4141 atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg
     4201 gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg
     4261 tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt
     4321 agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg
     4381 tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct
     4441 agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc
     4501 atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt
     4561 ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc
     4621 tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc
     4681 ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt
     4741 tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt
     4801 gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg
     4861 atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt
     4921 tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct
     4981 ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg
     5041 gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact
     5101 aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg
     5161 gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg taaattctgc
     5221 tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct
     5281 ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa
     5341 aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg
     5401 cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac
     5461 cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc
     5521 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
     5581 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
     5641 gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg
     5701 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc
     5761 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta
     5821 aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtacta gtgcttggat
     5881 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     5941 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgtaaac ttggtctgac
     6001 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6061 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6121 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6181 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6241 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6301 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6361 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6421 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6481 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6541 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6601 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6661 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6721 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6781 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6841 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6901 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6961 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
