LOCUS       sequence-229808-        3563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of norovirus antisense orientation toehold
            switches (A1) with a lacZ alpha subunit reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3563)
  AUTHORS   Ma D, Shen L, Wu K, Diehnelt CW, Green AA
  TITLE     Low-cost detection of norovirus using paper-based cell-free systems 
            and synbody-based viral enrichment.
  JOURNAL   Synth Biol (Oxf). 2018;3(1):ysy018. doi: 10.1093/synbio/ysy018. Epub
            2018 Sep 19.
  PUBMED    30370338
REFERENCE   2  (bases 1 to 3563)
  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: "Synth Biol 
            (Oxf)."; date: "2018"; volume: "3(1):ysy018. doi"; pages: " 
            10.1093/synbio/ysy018. Epub 2018 Sep 19"
COMMENT     Sequence Label: pDM_noro_A1_lacZA
FEATURES             Location/Qualifiers
     source          1..3563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             127..300
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(147..164)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(147..163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(156..178)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      310..356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(586..1401)
                     /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(586..1401)
                     /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     1309..1328
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1402..1493)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1511..2146)
                     /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(1656..1675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2344..3426)
                     /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     3388..3407
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3427..3504)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg tctgatgggt ccgcagccaa cctcgtccca gaggtcggac
       61 tttagaacag aggagataaa gatggacctc tgggaaacct ggcggcagcg caagaagatg
      121 cgtaaaatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa
      181 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      241 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      301 tggtaataat agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      361 aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
      421 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg acaagctgac
      481 gaccgggtct ccgcaagtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      541 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
      601 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
      661 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
      721 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
      781 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
      841 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
      901 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
      961 aaatacgcgg tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     1021 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     1081 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     1141 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     1201 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     1261 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     1321 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     1381 ttcccgttga atatggctca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggcat
     1501 gctagcgcag aaacgtccta gaagatgcca ggaggatact tagcagagag acaataaggc
     1561 cggagcgaag ccgtttttcc ataggctccg cccccctgac gaacatcacg aaatctgacg
     1621 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctga
     1681 tggctccctc ttgcgctctc ctgttcccgt cctgcggcgt ccgtgttgtg gtggaggctt
     1741 tacccaaatc accacgtccc gttccgtgta gacagttcgc tccaagctgg gctgtgtgca
     1801 agaacccccc gttcagcccg actgctgcgc cttatccggt aactatcatc ttgagtccaa
     1861 cccggaaaga cacgacaaaa cgccactggc agcagccatt ggtaactgag aattagtgga
     1921 tttagatatc gagagtcttg aagtggtggc ctaacagagg ctacactgaa aggacagtat
     1981 ttggtatctg cgctccacta aagccagtta ccaggttaag cagttcccca actgacttaa
     2041 ccttcgatca aaccgcctcc ccaggcggtt ttttcgttta cagagcagga gattacgacg
     2101 atcgtaaaag gatctcaaga agatccttta cggattcccg acaccatcac tctagatttc
     2161 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     2221 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     2281 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     2341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     2461 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     2521 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     2581 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     2641 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     2701 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     2761 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     2821 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     2881 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     2941 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     3001 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     3061 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     3121 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     3181 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     3241 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     3301 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     3361 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     3421 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     3481 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgt tgaaagtgac
     3541 caccgctcac gtgttgtgtg cgc
//
