LOCUS       sequence-178137-        5520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic gene circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  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 5520)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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: pZS2oxySp-GFP-proD-oxyR
FEATURES             Location/Qualifiers
     source          1..5520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             111..122
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(159..187)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     782..802
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      881..967
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2110..2196
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2741..3691)
                     /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      3739..3961
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4459..4553)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4584..5378)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4695..4714)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5305..5324
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ttcattatcc atcctccatc gccacgatag ttcatggcga taggtagaat agcaatgaac
       61 gattatccct atcaagcatt ctgactgata attgctcaca cgaattcatt aaagaggaga
      121 aaggtaccat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
      181 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
      241 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
      301 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
      361 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
      421 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
      481 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
      541 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
      601 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
      661 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
      721 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
      781 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gatctctaca
      841 aataacccgg gggatcccat ggtacgcgtg ctagaggcat caaataaaac gaaaggctca
      901 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
      961 gacaaatccg ccgccctaga cctagcacag ctaacaccac gtcgtcccta tctgctgccc
     1021 taggtctatg agtggttgct ggataacttt acgggcatgc ataaggctcg tataatatat
     1081 tcagggagac cacaacggtt tccctctaca aataattttg tttaactttg aattcttcac
     1141 acaggaaacc ggtaccatga atattcgtga tcttgagtac ctggtggcat tggctgaaca
     1201 ccgccatttt cggcgtgcgg cagattcctg ccacgttagc cagccgacgc ttagcgggca
     1261 aattcgtaag ctggaagatg agctgggcgt gatgttgctg gagcggacca gccgtaaagt
     1321 gttgttcacc caggcgggaa tgctgctggt ggatcaggcg cgtaccgtgc tgcgtgaggt
     1381 gaaagtcctt aaagagatgg caagccagca gggcgagacg atgtccggac cgctgcacat
     1441 tggtttgatt cccacagttg gaccgtacct gctaccgcat attatcccta tgctgcacca
     1501 gacctttcca aagctggaaa tgtatctgca tgaagcacag acccaccagt tactggcgca
     1561 actggacagc ggcaaactcg attgcgtgat cctcgcgctg gtgaaagaga gcgaagcatt
     1621 cattgaagtg ccgttgtttg atgagccaat gttgctggct atctatgaag atcacccgtg
     1681 ggcgaaccgc gaatgcgtac cgatggccga tctggcaggg gaaaaactgc tgatgctgga
     1741 agatggtcac tgtttgcgcg atcaggcaat gggtttctgt tttgaagccg gggcggatga
     1801 agatacacac ttccgcgcga ccagcctgga aactctgcgc aacatggtgg cggcaggtag
     1861 cgggatcact ttactgccag cgctggctgt gccgccggag cgcaaacgcg atggggttgt
     1921 ttatctgccg tgcattaagc cggaaccacg ccgcactatt ggcctggttt atcgtcctgg
     1981 ctcaccgctg cgcagccgct atgagcagct ggcagaggcc atccgcgcaa gaatggatgg
     2041 ccatttcgat aaagttttaa aacaggcggt ttaacccggg ggatcccatg gtacgcgtgc
     2101 tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2161 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac ctagggccta
     2221 gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt gttatcagaa
     2281 tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca
     2341 tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat
     2401 aagcagcatc gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa caagttgtct
     2461 caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg ttctattagg
     2521 tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag ctttgaatgc
     2581 accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt catctgtgca
     2641 tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt tgtttgttag
     2701 tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt ccgtatttag
     2761 ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac gaaccattga
     2821 gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg agctgaattt
     2881 ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag gaatctgatg
     2941 taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca agttcggtta
     3001 cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc
     3061 gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt attggtttca
     3121 aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt aacatgaact
     3181 taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt gttagttctt
     3241 ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta acatgttcca
     3301 gattatattt tatgaatttt tttaactgga aaagataagg caatatctct tcactaaaaa
     3361 ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa atctcaaagc
     3421 ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg gttgctttag
     3481 ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat tggttataag
     3541 tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg agtagtgcca
     3601 cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta atcgctagtt
     3661 catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg tgattttaat
     3721 cactatacca attgagatgg gctagtcaat gataattact agtccttttc ctttgagttg
     3781 tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct gctagaccct
     3841 ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg gttataattt
     3901 atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct gtgtataact
     3961 cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg tttgctcctc
     4021 tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc tcgggcaaat
     4081 cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac
     4141 attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc
     4201 cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc
     4261 tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag
     4321 ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag
     4381 actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg tcttactgtc
     4441 cctagtacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct
     4501 gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga
     4561 gctctcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
     4621 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     4681 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     4741 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     4801 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc
     4861 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     4921 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     4981 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     5041 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     5101 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     5161 gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
     5221 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     5281 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     5341 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     5401 tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     5461 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccgacgtc
//
