LOCUS       sequence-272409-        2921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 2BP dowsntream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2921)
  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 2921)
  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: pJBL703
FEATURES             Location/Qualifiers
     source          1..2921
                     /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"
     protein_bind    282..300
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      453..499
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(525..542)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(525..541)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(534..556)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(750..769)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     869..891
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(929..947)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1015..1119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1120..1188,1189..1980)
                     /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(1338..1357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2151..2739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2640..2659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2893..2910
                     /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 gtccctatca gtgatagaga
      301 cccacatact ctgatgatcc gagacggtcg ggtccagata ttcgtatctg tcgagtagag
      361 tgtgggctcg gatcattcat ggcaagagac ggtcgggtcc agatattcgt atctgtcgag
      421 tagagtgtgg gctcttgcca tgtgtatgtg ggtagcataa ccccttgggg cctctaaacg
      481 ggtcttgagg ggttttttga tccccgggta ccgagctcga attcactggc cgtcgtttta
      541 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      601 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      661 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      721 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      781 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
      841 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
      901 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
      961 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1021 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1081 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1141 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1201 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1261 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1321 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1381 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1441 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1501 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1561 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1621 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1681 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1741 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1801 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1861 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1921 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1981 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2041 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2101 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2161 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2221 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2281 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2341 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2401 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2461 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2521 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2581 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2641 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2701 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2761 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2821 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2881 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag a
//
