LOCUS       sequence-272393-        2936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MphR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the mphO sequence 5BP downstream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2936)
  AUTHORS   Jung JK, Alam KK, Verosloff MS, Capdevila DA, Desmau M, Clauer PR, 
            Lee JW, Nguyen PQ, Pasten PA, Matiasek SJ, Gaillard JF, Giedroc DP, 
            Collins JJ, Lucks JB
  TITLE     Cell-free biosensors for rapid detection of water contaminants.
  JOURNAL   Nat Biotechnol. 2020 Jul 6. pii: 10.1038/s41587-020-0571-7. doi: 
            10.1038/s41587-020-0571-7.
  PUBMED    32632301
REFERENCE   2  (bases 1 to 2936)
  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: "Nat 
            Biotechnol. 2020 Jul 6. pii: 10.1038/s41587-020-0571-7. doi: 
            10.1038/s41587-020-0571-7."
COMMENT     Sequence Label: pJBL716
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        263..281
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      471..517
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(540..557)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(540..556)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(549..571)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(765..784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     884..906
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(944..962)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1030..1134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1135..1203,1204..1995)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1353..1372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2166..2754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2655..2674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2908..2925
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gataatacga ctcactatag gagggaatat aaccgacgtg
      301 actgttacat ttaggtggcc cacatactct gatgatccga gacggtcggg tccagatatt
      361 cgtatctgtc gagtagagtg tgggctcgga tcattcatgg caagagacgg tcgggtccag
      421 atattcgtat ctgtcgagta gagtgtgggc tcttgccatg tgtatgtggg tagcataacc
      481 ccttggggcc tctaaacggg tcttgagggg ttttttgccc gggtaccgag ctcgaattca
      541 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
      601 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
      661 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
      721 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
      781 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
      841 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
      901 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
      961 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1021 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1081 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1141 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1201 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1261 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1321 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1381 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1441 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1501 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1561 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1621 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1681 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1741 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1801 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1861 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1921 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1981 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2041 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2101 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2161 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2221 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2281 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2341 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2401 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2461 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2521 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2581 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2641 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2701 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2761 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2821 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2881 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
