LOCUS       sequence-235986-        6454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of norovirus antisense orientation toehold
            switches (A2) with a lacZ reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6454)
  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 6454)
  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_lacZ
FEATURES             Location/Qualifiers
     source          1..6454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     926..948
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     940..957
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     941..957
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1083..1102)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1084..1102)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1162..1239
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1240..2322
                     /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
                     ALVDSLMQLARQVSRLESGQ"
     primer_bind     complement(1259..1278)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      2520..3155
                     /direction=RIGHT
                     /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     2991..3010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3173..3264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3265..4080
                     /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             3265..4080
                     /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     complement(3338..3357)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(4310..4356)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(4357..6454,1..977))
                     /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"
ORIGIN
        1 gcttcaatca gcgtgccgtc ggcggtgtgc agttcaacca ccgcacgata gagattcggg
       61 atttcggcgc tccacagttt cgggttttcg acgttcagac gtagtgtgac gcgatcggca
      121 taaccaccac gctcatcgat aatttcaccg ccgaaaggcg cggtgccgct ggcgacctgc
      181 gtttcaccct gccataaaga aactgttacc cgtaggtagt cacgcaactc gccgcacatc
      241 tgaacttcag cctccagtac agcgcggctg aaatcatcat taaagcgagt ggcaacatgg
      301 aaatcgctga tttgtgtagt cggtttatgc agcaacgaga cgtcacggaa aatgccgctc
      361 atccgccaca tatcctgatc ttccagataa ctgccgtcac tccagcgcag caccatcacc
      421 gcgaggcggt tttctccggc gcgtaaaaat gcgctcaggt caaattcaga cggcaaacga
      481 ctgtcctggc cgtaaccgac ccagcgcccg ttgcaccaca gatgaaacgc cgagttaacg
      541 ccatcaaaaa taattcgcgt ctggccttcc tgtagccagc tttcatcaac attaaatgtg
      601 agcgagtaac aacccgtcgg attctccgtg ggaacaaacg gcggattgac cgtaatggga
      661 taggtcacgt tggtgtagat gggcgcatcg taaccgtgca tctgccagtt tgaggggacg
      721 acgacagtat cggcctcagg aagatcgcac tccagccagc tttccggcac cgcttctggt
      781 gccggaaacc aggcaaagcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      841 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      901 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
      961 tccgtaatca tggtcatttt acgcatcttc ttgcgctgcc gccaggttcc cagttttgtc
     1021 atctttatct cctctgttct aaagtccaca aaactgggag ccagattgcg atcgccctcc
     1081 caccctatag tgagtcgtat tagcgcacac aacacgtgag cggtggtcac tttcaacata
     1141 agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca
     1201 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     1261 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     1321 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     1381 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     1441 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     1501 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     1561 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     1621 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     1681 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     1741 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     1801 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     1861 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     1921 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     1981 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggac
     2041 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
     2101 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     2161 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     2221 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggtcga ttcattaatg
     2281 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2341 aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc
     2401 agtcagctcc ttccggtggg cgcggggcat gactaacatg agaattacaa cttatatcgt
     2461 atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagagtgat
     2521 ggtgtcggga atccgtaaag gatcttcttg agatcctttt acgatcgtcg taatctcctg
     2581 ctctgtaaac gaaaaaaccg cctggggagg cggtttgatc gaaggttaag tcagttgggg
     2641 aactgcttaa cctggtaact ggctttagtg gagcgcagat accaaatact gtcctttcag
     2701 tgtagcctct gttaggccac cacttcaaga ctctcgatat ctaaatccac taattctcag
     2761 ttaccaatgg ctgctgccag tggcgttttg tcgtgtcttt ccgggttgga ctcaagatga
     2821 tagttaccgg ataaggcgca gcagtcgggc tgaacggggg gttcttgcac acagcccagc
     2881 ttggagcgaa ctgtctacac ggaacgggac gtggtgattt gggtaaagcc tccaccacaa
     2941 cacggacgcc gcaggacggg aacaggagag cgcaagaggg agccatcagg gggaaacgcc
     3001 tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga
     3061 tgttcgtcag gggggcggag cctatggaaa aacggcttcg ctccggcctt attgtctctc
     3121 tgctaagtat cctcctggca tcttctagga cgtttctgcg ctagcatgcc tatttgttta
     3181 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3241 caataatatt gaaaaaggaa gagtatgagc catattcaac gggaaacgtc ttgctctagg
     3301 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     3361 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     3421 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     3481 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     3541 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
     3601 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     3661 tcgattcctg tttgtaattg tccttttaac agcgaccgcg tatttcgtct cgctcaggcg
     3721 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     3781 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     3841 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     3901 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     3961 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     4021 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     4081 gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4141 tccgcgcaca tttccccgaa aagtgccact tgcggagacc cggtcgtcag cttgtcgtcg
     4201 gttcagggca gggtcgttaa atagccgctt atgtctattg ctggtttacc ggtttattga
     4261 ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg aggtttcagc aaaaaacccc
     4321 tcaagacccg tttagaggcc ccaaggggtt atgctattat ttttgacacc agaccaactg
     4381 gtaatggtag cgaccggcgc tcagctggaa ttccgccgat actgacgggc tccaggagtc
     4441 gtcgccacca atccccatat ggaaaccgtc gatattcagc catgtgcctt cttccgcgtg
     4501 cagcagatgg cgatggctgg tttccatcag ttgctgttga ctgtagcggc tgatgttgaa
     4561 ctggaagtcg ccgcgccact ggtgtgggcc ataattcaat tcgcgcgtcc cgcagcgcag
     4621 accgttttcg ctcgggaaga cgtacggggt atacatgtct gacaatggca gatcccagcg
     4681 gtcaaaacag gcggcagtaa ggcggtcggg atagttttct tgcggcccta atccgagcca
     4741 gtttacccgc tctgctacct gcgccagctg gcagttcagg ccaatccgcg ccggatgcgg
     4801 tgtatcgctc gccacttcaa catcaacggt aatcgccatt tgaccactac catcaatccg
     4861 gtaggttttc cggctgataa ataaggtttt cccctgatgc tgccacgcgt gagcggtcgt
     4921 aatcagcacc gcatcagcaa gtgtatctgc cgtgcactgc aacaacgctg cttcggcctg
     4981 gtaatggccc gccgccttcc agcgttcgac ccaggcgtta gggtcaatgc gggtcgcttc
     5041 acttacgcca atgtcgttat ccagcggtgc acgggtgaac tgatcgcgca gcggcgtcag
     5101 cagttgtttt ttatcgccaa tccacatctg tgaaagaaag cctgactggc ggttaaattg
     5161 ccaacgctta ttacccagct cgatgcaaaa atccatttcg ctggtggtca gatgcgggat
     5221 ggcgtgggac gcggcgggga gcgtcacact gaggttttcc gccagacgcc actgctgcca
     5281 ggcgctgatg tgcccggctt ctgaccatgc ggtcgcgttc ggttgcacta cgcgtactgt
     5341 gagccagagt tgcccggcgc tctccggctg cggtagttca ggcagttcaa tcaactgttt
     5401 accttgtgga gcgacatcca gaggcacttc accgcttgcc agcggcttac catccagcgc
     5461 caccatccag tgcaggagct cgttatcgct atgacggaac aggtattcgc tggtcacttc
     5521 gatggtttgc ccggataaac ggaactggaa aaactgctgc tggtgttttg cttccgtcag
     5581 cgctggatgc ggcgtgcggt cggcaaagac cagaccgttc atacagaact ggcgatcgtt
     5641 cggcgtatcg ccaaaatcac cgccgtaagc cgaccacggg ttgccgtttt catcatattt
     5701 aatcagcgac tgatccaccc agtcccagac gaagccgccc tgtaaacggg gatactgacg
     5761 aaacgcctgc cagtatttag cgaaaccgcc aagactgtta cccatcgcgt gggcgtattc
     5821 gcaaaggatc agcgggcgcg tctctccagg tagcgaaagc cattttttga tggaccattt
     5881 cggcacagcc gggaagggct ggtcttcatc cacgcgcgcg tacatcgggc aaataatatc
     5941 ggtggccgtg gtgtcggctc cgccgccttc atactgcacc gggcgggaag gatcgacaga
     6001 tttgatccag cgatacagcg cgtcgtgatt agcgccgtgg cctgattcat tccccagcga
     6061 ccagatgatc acactcgggt gattacgatc gcgctgcacc attcgcgtta cgcgttcgct
     6121 catcgccggt agccagcgcg gatcatcggt cagacgattc attggcacca tgccgtgggt
     6181 ttcaatattg gcttcatcca ccacatacag gccgtagcgg tcgcacagcg tgtaccacag
     6241 cggatggttc ggataatgcg aacagcgcac ggcgttaaag ttgttctgct tcatcagcag
     6301 gatatcctgc accatcgtct gctcatccat gacctgacca tgcagaggat gatgctcgtg
     6361 acggttaacg cctcgaatca gcaacggctt gccgttcagc agcagcagac cattttcaat
     6421 ccgcacctcg cggaaaccga catcgcaggc ttct
//
