LOCUS       sequence-297854-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses QacR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the qacO 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: pJBL720
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..129
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    246..267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(282..287,288..305,306..312)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    320..336
                     /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     325..347
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     344..360
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     344..360
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        403..421
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      605..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(677..694)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(677..693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(686..708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(902..921)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1021..1043
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1081..1099)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1167..1271
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1272..1340,1341..2132)
                     /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(1490..1509)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2303..2891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2792..2811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      121 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
      181 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
      241 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      301 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      361 catgattacg ccaagcttgc atgcctgcag gtcgactcta gataatacga ctcactatag
      421 gaggcttata gaccgatcgc acggtctata agcccacata ctctgatgat ccgagacggt
      481 cgggtccaga tattcgtatc tgtcgagtag agtgtgggct cggatcattc atggcaagag
      541 acggtcgggt ccagatattc gtatctgtcg agtagagtgt gggctcttgc catgtgtatg
      601 tgggtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gatccccggg
      661 taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
      841 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
      901 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
      961 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     1021 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     1081 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     1141 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1201 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1261 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1321 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1381 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1441 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1501 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1561 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1621 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1681 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1741 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1801 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1861 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1921 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1981 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2041 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2101 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2161 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2221 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgc
//
