LOCUS       sequence-297846-        2924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 5BP dowsntream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2924)
  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 2924)
  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: pJBL705
FEATURES             Location/Qualifiers
     source          1..2924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    265..286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(301..306,307..324,325..331)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    339..355
                     /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     344..366
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     363..379
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     363..379
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        422..440
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    444..462
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      615..661
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(687..704)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(687..703)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(696..718)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(912..931)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1031..1053
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1091..1109)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1177..1281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1282..1350,1351..2142)
                     /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(1500..1519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2313..2901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2802..2821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
       61 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      121 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      181 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      241 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      301 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      361 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag
      421 ataatacgac tcactatagg aggtccctat cagtgataga gacccacata ctctgatgat
      481 ccgagacggt cgggtccaga tattcgtatc tgtcgagtag agtgtgggct cggatcattc
      541 atggcaagag acggtcgggt ccagatattc gtatctgtcg agtagagtgt gggctcttgc
      601 catgtgtatg tgggtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      661 gatccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
      721 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      781 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      841 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
      901 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
      961 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     1021 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     1081 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1141 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1201 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1261 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1321 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1381 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1441 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1501 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1561 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1621 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1681 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1741 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1801 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     1861 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1921 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1981 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2041 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2101 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2161 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2221 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2281 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2341 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2401 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2461 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2521 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2581 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2641 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2701 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2761 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2821 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2881 ggggggcgga gcctatggaa aaacgccagc aacgcggcct tttt
//
