LOCUS       sequence-229807-        3562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of norovirus antisense orientation toehold
            switches (A2) with a lacZ alpha subunit reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3562)
  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 3562)
  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_A2_lacZA
FEATURES             Location/Qualifiers
     source          1..3562
                     /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             126..299
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(146..163)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(146..162)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(155..177)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      309..355
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(585..1400)
                     /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(585..1400)
                     /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     1308..1327
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1401..1492)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1510..2145)
                     /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(1655..1674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2343..3425)
                     /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     3387..3406
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3426..3503)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg tgggagggcg atcgcaatct ggctcccagt tttgtggact
       61 ttagaacaga ggagataaag atgacaaaac tgggaacctg gcggcagcgc aagaagatgc
      121 gtaaaatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      181 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      241 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      301 ggtaataata gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      361 acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
      421 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga caagctgacg
      481 accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
      541 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
      601 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
      661 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
      721 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
      781 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
      841 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
      901 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
      961 aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1021 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1081 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1141 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1201 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1261 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     1321 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt
     1381 tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1441 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggcatg
     1501 ctagcgcaga aacgtcctag aagatgccag gaggatactt agcagagaga caataaggcc
     1561 ggagcgaagc cgtttttcca taggctccgc ccccctgacg aacatcacga aatctgacgc
     1621 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgat
     1681 ggctccctct tgcgctctcc tgttcccgtc ctgcggcgtc cgtgttgtgg tggaggcttt
     1741 acccaaatca ccacgtcccg ttccgtgtag acagttcgct ccaagctggg ctgtgtgcaa
     1801 gaaccccccg ttcagcccga ctgctgcgcc ttatccggta actatcatct tgagtccaac
     1861 ccggaaagac acgacaaaac gccactggca gcagccattg gtaactgaga attagtggat
     1921 ttagatatcg agagtcttga agtggtggcc taacagaggc tacactgaaa ggacagtatt
     1981 tggtatctgc gctccactaa agccagttac caggttaagc agttccccaa ctgacttaac
     2041 cttcgatcaa accgcctccc caggcggttt tttcgtttac agagcaggag attacgacga
     2101 tcgtaaaagg atctcaagaa gatcctttac ggattcccga caccatcact ctagatttca
     2161 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     2221 ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     2281 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     2341 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2401 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     2461 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     2521 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     2581 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     2641 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     2701 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     2761 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     2821 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     2881 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     2941 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     3001 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     3061 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     3121 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     3181 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     3241 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     3301 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     3361 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     3421 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     3481 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgtt gaaagtgacc
     3541 accgctcacg tgttgtgtgc gc
//
