LOCUS       sequence-224529-        3554 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 3554)
  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 3554)
  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..3554
                     /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             152..325
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(172..189)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(172..188)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(181..203)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      335..381
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(611..1426)
                     /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(611..1426)
                     /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     1334..1353
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1427..1518)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1536..2171)
                     /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(1681..1700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2369..3451)
                     /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     3413..3432
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3452..3529)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctagtaaatt cgcgtttcta cggtagccgg gcgctaatac gactcactat agggtctgat
       61 gggtccgcag ccaacctcgt cccagaggtc ggactttaga acagaggaga taaagatgga
      121 cctctgggaa acctggcggc agcgcaagaa gatgaccatg attacggatt cactggccgt
      181 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      241 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      301 acagttgcgc agcctgaatg gcgaatggta ataatagcat aaccccttgg ggcctctaaa
      361 cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac ggtcacactg
      421 cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc
      481 tgccctgaac cgacgacaag ctgacgaccg ggtctccgca agtggcactt ttcggggaaa
      541 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      601 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
      661 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
      721 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
      781 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
      841 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
      901 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
      961 ttcgtgattg cgcctgagcg agacgaaata cgcggtcgct gttaaaagga caattacaaa
     1021 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     1081 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     1141 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     1201 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     1261 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     1321 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     1381 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcatactc ttcctttttc
     1441 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1501 tttagaaaaa taaacaaata ggcatgctag cgcagaaacg tcctagaaga tgccaggagg
     1561 atacttagca gagagacaat aaggccggag cgaagccgtt tttccatagg ctccgccccc
     1621 ctgacgaaca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat
     1681 aaagatacca ggcgtttccc cctgatggct ccctcttgcg ctctcctgtt cccgtcctgc
     1741 ggcgtccgtg ttgtggtgga ggctttaccc aaatcaccac gtcccgttcc gtgtagacag
     1801 ttcgctccaa gctgggctgt gtgcaagaac cccccgttca gcccgactgc tgcgccttat
     1861 ccggtaacta tcatcttgag tccaacccgg aaagacacga caaaacgcca ctggcagcag
     1921 ccattggtaa ctgagaatta gtggatttag atatcgagag tcttgaagtg gtggcctaac
     1981 agaggctaca ctgaaaggac agtatttggt atctgcgctc cactaaagcc agttaccagg
     2041 ttaagcagtt ccccaactga cttaaccttc gatcaaaccg cctccccagg cggttttttc
     2101 gtttacagag caggagatta cgacgatcgt aaaaggatct caagaagatc ctttacggat
     2161 tcccgacacc atcactctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     2221 tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa
     2281 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     2341 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2401 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2461 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     2521 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     2581 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     2641 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     2701 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     2761 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     2821 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     2881 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     2941 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     3001 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     3061 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     3121 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     3181 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     3241 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     3301 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     3361 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     3421 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     3481 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     3541 cgctctccct tatg
//
