LOCUS       sequence-297855-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TtgR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the ttgO sequence 5BP downstream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2933)
  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 2933)
  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: pJBL722
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        252..270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      454..500
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(526..543)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(526..542)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(535..557)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(751..770)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     870..892
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(930..948)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1016..1120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1121..1189,1190..1981)
                     /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(1339..1358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2152..2740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2641..2660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2894..2911
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg
      241 tcgactctag ataatacgac tcactatagg aggtatttac aaacaaccat gaatgtaagt
      301 acccacatac tctgatgatc cgagacggtc gggtccagat attcgtatct gtcgagtaga
      361 gtgtgggctc ggatcattca tggcaagaga cggtcgggtc cagatattcg tatctgtcga
      421 gtagagtgtg ggctcttgcc atgtgtatgt gggtagcata accccttggg gcctctaaac
      481 gggtcttgag gggttttttg atccccgggt accgagctcg aattcactgg ccgtcgtttt
      541 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
      601 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
      661 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
      721 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      781 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      841 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      901 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      961 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     1021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1081 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1141 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1201 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1261 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1321 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     1381 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1441 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1501 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1561 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1621 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     1681 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1741 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1801 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1861 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1921 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1981 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2281 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2821 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2881 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tac
//
