LOCUS       sequence-297860-        2931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 12BP dowsntream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2931)
  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 2931)
  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: pJBL709
FEATURES             Location/Qualifiers
     source          1..2931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..79
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    196..217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(232..237,238..255,256..262)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    270..286
                     /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     275..297
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     294..310
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     294..310
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     353..372
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        353..371
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    382..400
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      553..599
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(625..642)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(625..641)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(634..656)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(850..869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     969..991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1029..1047)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1115..1219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1220..1288,1289..2080)
                     /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(1438..1457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2251..2839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2740..2759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
       61 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      121 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      181 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      241 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      301 cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta gataatacga
      361 ctcactatag ggagaccaca atccctatca gtgatagaga cccacatact ctgatgatcc
      421 gagacggtcg ggtccagata ttcgtatctg tcgagtagag tgtgggctcg gatcattcat
      481 ggcaagagac ggtcgggtcc agatattcgt atctgtcgag tagagtgtgg gctcttgcca
      541 tgtgtatgtg ggtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttga
      601 tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
      661 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      721 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      781 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
      841 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
      901 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
      961 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     1021 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     1081 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1141 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1201 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1261 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1321 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1381 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1441 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1501 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1561 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1621 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1681 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1741 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1801 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1861 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1921 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1981 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     2041 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     2101 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2161 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2221 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttctctgc gcgtaatctg
     2281 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2341 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2521 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2581 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2641 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2701 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2761 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2821 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2881 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg a
//
