LOCUS       sequence-146082-        6455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Toehold switch sensor to detect Zika Virus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6455)
  AUTHORS   Pardee K, Green AA, Takahashi MK, Braff D, Lambert G, Lee JW, 
            Ferrante T, Ma D, Donghia N, Fan M, Daringer NM, Bosch I, Dudley DM,
            O'Connor DH, Gehrke L, Collins JJ
  TITLE     Rapid, Low-Cost Detection of Zika Virus Using Programmable 
            Biomolecular Components.
  JOURNAL   Cell. 2016 May 6. pii: S0092-8674(16)30505-0. doi: 
            10.1016/j.cell.2016.04.059.
  PUBMED    27160350
REFERENCE   2  (bases 1 to 6455)
  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: "Cell. 2016 
            May 6. pii: S0092-8674(16)30505-0. doi: 10.1016/j.cell.2016.04.059."
COMMENT     Sequence Label: ZIKV_Sensor_32B_LacZ
FEATURES             Location/Qualifiers
     source          1..6455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..54
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        35..53
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             161..3235
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(181..198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(181..197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(190..212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3236..3282
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3512..4327)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3512..4327)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4235..4254
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4328..4419)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4437..5072)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(4582..4601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5270..6352)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6314..6333
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6353..6430)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gtagtctgcc agtggttcgc gatatagctc gcgctaatac gactcactat agggctggga
       61 tcaagtacat gtagtgcgcc acgagcaaaa ggactttaga acagaggaga taaagatgtt
      121 ttgctcgtgt aacctggcgg cagcgcaaaa gatgcgtaaa atgaccatga ttacggattc
      181 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      241 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      301 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc
      361 agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt
      421 cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc
      481 cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac
      541 atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt
      601 taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc ggttacggcc aggacagtcg
      661 tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc ggagaaaacc gcctcgcggt
      721 gatggtgctg cgctggagtg acggcagtta tctggaagat caggatatgt ggcggatgag
      781 cggcattttc cgtgacgtct cgttgctgca taaaccgact acacaaatca gcgatttcca
      841 tgttgccact cgctttaatg atgatttcag ccgcgctgta ctggaggctg aagttcagat
      901 gtgcggcgag ttgcgtgact acctacgggt aacagtttct ttatggcagg gtgaaacgca
      961 ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc gatgagcgtg gtggttatgc
     1021 cgatcgcgtc acactacgtc tgaacgtcga aaacccgaaa ctgtggagcg ccgaaatccc
     1081 gaatctctat cgtgcggtgg ttgaactgca caccgccgac ggcacgctga ttgaagcaga
     1141 agcctgcgat gtcggtttcc gcgaggtgcg gattgaaaat ggtctgctgc tgctgaacgg
     1201 caagccgttg ctgattcgag gcgttaaccg tcacgagcat catcctctgc atggtcaggt
     1261 catggatgag cagacgatgg tgcaggatat cctgctgatg aagcagaaca actttaacgc
     1321 cgtgcgctgt tcgcattatc cgaaccatcc gctgtggtac acgctgtgcg accgctacgg
     1381 cctgtatgtg gtggatgaag ccaatattga aacccacggc atggtgccaa tgaatcgtct
     1441 gaccgatgat ccgcgctggc taccggcgat gagcgaacgc gtaacgcgaa tggtgcagcg
     1501 cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg aatgaatcag gccacggcgc
     1561 taatcacgac gcgctgtatc gctggatcaa atctgtcgat ccttcccgcc cggtgcagta
     1621 tgaaggcggc ggagccgaca ccacggccac cgatattatt tgcccgatgt acgcgcgcgt
     1681 ggatgaagac cagcccttcc cggctgtgcc gaaatggtcc atcaaaaaat ggctttcgct
     1741 acctggagag acgcgcccgc tgatcctttg cgaatacgcc cacgcgatgg gtaacagtct
     1801 tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat ccccgtttac agggcggctt
     1861 cgtctgggac tgggtggatc agtcgctgat taaatatgat gaaaacggca acccgtggtc
     1921 ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc cagttctgta tgaacggtct
     1981 ggtctttgcc gaccgcacgc cgcatccagc gctgacggaa gcaaaacacc agcagcagtt
     2041 tttccagttc cgtttatccg ggcaaaccat cgaagtgacc agcgaatacc tgttccgtca
     2101 tagcgataac gagctcctgc actggatggt ggcgctggat ggtaagccgc tggcaagcgg
     2161 tgaagtgcct ctggatgtcg ctccacaagg taaacagttg attgaactgc ctgaactacc
     2221 gcagccggag agcgccgggc aactctggct cacagtacgc gtagtgcaac cgaacgcgac
     2281 cgcatggtca gaagccgggc acatcagcgc ctggcagcag tggcgtctgg cggaaaacct
     2341 cagtgtgacg ctccccgccg cgtcccacgc catcccgcat ctgaccacca gcgaaatgga
     2401 tttttgcatc gagctgggta ataagcgttg gcaatttaac cgccagtcag gctttctttc
     2461 acagatgtgg attggcgata aaaaacaact gctgacgccg ctgcgcgatc agttcacccg
     2521 tgcaccgctg gataacgaca ttggcgtaag tgaagcgacc cgcattgacc ctaacgcctg
     2581 ggtcgaacgc tggaaggcgg cgggccatta ccaggccgaa gcagcgttgt tgcagtgcac
     2641 ggcagataca cttgctgatg cggtgctgat tacgaccgct cacgcgtggc agcatcaggg
     2701 gaaaacctta tttatcagcc ggaaaaccta ccggattgat ggtagtggtc aaatggcgat
     2761 taccgttgat gttgaagtgg cgagcgatac accgcatccg gcgcggattg gcctgaactg
     2821 ccagctggcg caggtagcag agcgggtaaa ctggctcgga ttagggccgc aagaaaacta
     2881 tcccgaccgc cttactgccg cctgttttga ccgctgggat ctgccattgt cagacatgta
     2941 taccccgtac gtcttcccga gcgaaaacgg tctgcgctgc gggacgcgcg aattgaatta
     3001 tggcccacac cagtggcgcg gcgacttcca gttcaacatc agccgctaca gtcaacagca
     3061 actgatggaa accagccatc gccatctgct gcacgcggaa gaaggcacat ggctgaatat
     3121 cgacggtttc catatgggga ttggtggcga cgactcctgg agcccgtcag tatcggcgga
     3181 attccagctg agcgccggtc gctaccatta ccagttggtc tggtgtcaaa aataatagca
     3241 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt
     3301 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca
     3361 taagcggcta tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc gggtctccgc
     3421 aagtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3481 ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa
     3541 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     3601 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     3661 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     3721 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     3781 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     3841 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcggtcgc
     3901 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     3961 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     4021 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     4081 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     4141 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     4201 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     4261 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
     4321 ggctcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4381 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggcatgcta gcgcagaaac
     4441 gtcctagaag atgccaggag gatacttagc agagagacaa taaggccgga gcgaagccgt
     4501 ttttccatag gctccgcccc cctgacgaac atcacgaaat ctgacgctca aatcagtggt
     4561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctgatggc tccctcttgc
     4621 gctctcctgt tcccgtcctg cggcgtccgt gttgtggtgg aggctttacc caaatcacca
     4681 cgtcccgttc cgtgtagaca gttcgctcca agctgggctg tgtgcaagaa ccccccgttc
     4741 agcccgactg ctgcgcctta tccggtaact atcatcttga gtccaacccg gaaagacacg
     4801 acaaaacgcc actggcagca gccattggta actgagaatt agtggattta gatatcgaga
     4861 gtcttgaagt ggtggcctaa cagaggctac actgaaagga cagtatttgg tatctgcgct
     4921 ccactaaagc cagttaccag gttaagcagt tccccaactg acttaacctt cgatcaaacc
     4981 gcctccccag gcggtttttt cgtttacaga gcaggagatt acgacgatcg taaaaggatc
     5041 tcaagaagat cctttacgga ttcccgacac catcactcta gatttcagtg caatttatct
     5101 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt
     5161 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     5221 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     5281 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5341 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     5401 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     5461 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     5521 cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg
     5581 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     5641 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     5701 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     5761 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     5821 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     5881 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     5941 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     6001 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     6061 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     6121 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     6181 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     6241 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     6301 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     6361 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     6421 cgatggtgtc cgggatctcg acgctctccc ttatg
//
