LOCUS       sequence-297856-        2927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CadC-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the cadO sequence 5BP downstream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2927)
  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 2927)
  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: pJBL729
FEATURES             Location/Qualifiers
     source          1..2927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..116
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    233..254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(269..274,275..292,293..299)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    307..323
                     /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     312..334
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     331..347
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     331..347
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        390..408
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      586..632
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(658..675)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(658..674)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(667..689)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(883..902)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1002..1024
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1062..1080)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1148..1252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1253..1321,1322..2113)
                     /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(1471..1490)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2284..2872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2773..2792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
       61 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      121 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      181 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      241 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      301 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      361 agcttgcatg cctgcaggtc gactctagat aatacgactc actataggag gctcaaataa
      421 atatttgaat gaacccacat actctgatga tccgagacgg tcgggtccag atattcgtat
      481 ctgtcgagta gagtgtgggc tcggatcatt catggcaaga gacggtcggg tccagatatt
      541 cgtatctgtc gagtagagtg tgggctcttg ccatgtgtat gtgggtagca taaccccttg
      601 gggcctctaa acgggtcttg aggggttttt tgatccccgg gtaccgagct cgaattcact
      661 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
      721 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
      781 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
      841 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
      901 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      961 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     1021 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     1081 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     1141 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1201 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1261 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1321 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1381 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1441 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1501 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1561 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1621 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1681 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1741 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1801 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1861 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1921 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1981 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2041 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2101 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2401 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2521 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2581 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2641 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2701 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2761 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2821 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2881 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctc
//
