LOCUS       sequence-297844-        2923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 4BP dowsntream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2923)
  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 2923)
  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: pJBL704
FEATURES             Location/Qualifiers
     source          1..2923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    299..320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(335..340,341..358,359..365)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    373..389
                     /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     378..400
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     397..413
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     397..413
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     456..475
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        456..474
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    477..495
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      648..694
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(720..737)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(720..736)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(729..751)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(945..964)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1064..1086
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1124..1142)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1210..1314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1315..1383,1384..2175)
                     /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(1533..1552)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2346..2923,1..11)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2835..2854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
       61 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      181 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      241 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      301 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
      361 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
      421 acgccaagct tgcatgcctg caggtcgact ctagataata cgactcacta tagggatccc
      481 tatcagtgat agagacccac atactctgat gatccgagac ggtcgggtcc agatattcgt
      541 atctgtcgag tagagtgtgg gctcggatca ttcatggcaa gagacggtcg ggtccagata
      601 ttcgtatctg tcgagtagag tgtgggctct tgccatgtgt atgtgggtag cataacccct
      661 tggggcctct aaacgggtct tgaggggttt tttgatcccc gggtaccgag ctcgaattca
      721 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
      781 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
      841 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
      901 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
      961 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     1021 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1081 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     1141 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1201 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1261 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1321 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1381 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1441 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1501 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1561 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1621 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1681 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1741 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1801 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1861 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1921 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1981 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2041 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2101 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2161 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2221 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2281 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2341 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2401 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2461 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2521 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2581 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2641 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2701 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2761 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2821 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2881 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc gga
//
