LOCUS       sequence-224532-        3553 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 3553)
  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 3553)
  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..3553
                     /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             151..324
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(171..188)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(171..187)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(180..202)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      334..380
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(610..1425)
                     /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(610..1425)
                     /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     1333..1352
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1426..1517)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1535..2170)
                     /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(1680..1699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2368..3450)
                     /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     3412..3431
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3451..3528)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctagtaaatt cgcgtttcta cggtagccgg gcgctaatac gactcactat agggtgggag
       61 ggcgatcgca atctggctcc cagttttgtg gactttagaa cagaggagat aaagatgaca
      121 aaactgggaa cctggcggca gcgcaagaag atgaccatga ttacggattc actggccgtc
      181 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      241 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      301 cagttgcgca gcctgaatgg cgaatggtaa taatagcata accccttggg gcctctaaac
      361 gggtcttgag gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc
      421 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
      481 gccctgaacc gacgacaagc tgacgaccgg gtctccgcaa gtggcacttt tcggggaaat
      541 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      601 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
      661 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
      721 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
      781 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
      841 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
      901 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
      961 tcgtgattgc gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac aattacaaac
     1021 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     1081 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     1141 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     1201 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     1261 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     1321 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     1381 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct tcctttttca
     1441 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1501 ttagaaaaat aaacaaatag gcatgctagc gcagaaacgt cctagaagat gccaggagga
     1561 tacttagcag agagacaata aggccggagc gaagccgttt ttccataggc tccgcccccc
     1621 tgacgaacat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
     1681 aagataccag gcgtttcccc ctgatggctc cctcttgcgc tctcctgttc ccgtcctgcg
     1741 gcgtccgtgt tgtggtggag gctttaccca aatcaccacg tcccgttccg tgtagacagt
     1801 tcgctccaag ctgggctgtg tgcaagaacc ccccgttcag cccgactgct gcgccttatc
     1861 cggtaactat catcttgagt ccaacccgga aagacacgac aaaacgccac tggcagcagc
     1921 cattggtaac tgagaattag tggatttaga tatcgagagt cttgaagtgg tggcctaaca
     1981 gaggctacac tgaaaggaca gtatttggta tctgcgctcc actaaagcca gttaccaggt
     2041 taagcagttc cccaactgac ttaaccttcg atcaaaccgc ctccccaggc ggttttttcg
     2101 tttacagagc aggagattac gacgatcgta aaaggatctc aagaagatcc tttacggatt
     2161 cccgacacca tcactctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     2221 cagccccata cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag
     2281 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     2341 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2401 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2461 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     2521 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     2581 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     2641 taccgagatg tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     2701 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     2761 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     2821 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     2881 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     2941 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     3001 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     3061 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     3121 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     3181 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     3241 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     3301 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     3361 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     3421 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     3481 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     3541 gctctccctt atg
//
