LOCUS       sequence-297838-        2929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 10BP dowsntream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2929)
  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 2929)
  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: pJBL708
FEATURES             Location/Qualifiers
     source          1..2929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..82
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    199..220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(235..240,241..258,259..265)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    273..289
                     /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     278..300
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     297..313
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     297..313
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     356..375
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        356..374
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    383..401
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      554..600
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(626..643)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(626..642)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(635..657)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(851..870)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     970..992
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1030..1048)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1116..1220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1221..1289,1290..2081)
                     /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(1439..1458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2252..2840
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2741..2760
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
       61 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      121 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      181 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      241 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      301 aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagataata
      361 cgactcacta tagggagacc actccctatc agtgatagag acccacatac tctgatgatc
      421 cgagacggtc gggtccagat attcgtatct gtcgagtaga gtgtgggctc ggatcattca
      481 tggcaagaga cggtcgggtc cagatattcg tatctgtcga gtagagtgtg ggctcttgcc
      541 atgtgtatgt gggtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      601 atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      661 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      721 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      781 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
      841 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
      901 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
      961 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     1021 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1081 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1141 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1201 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1261 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1321 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1381 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1441 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1501 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1561 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1621 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1681 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1741 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     1801 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1861 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1921 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1981 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2041 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2101 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2161 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2221 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2281 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2341 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2401 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2461 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2521 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2581 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2641 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2701 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2761 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2821 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2881 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgt
//
