LOCUS       sequence-297861-        2929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HucR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the hucO sequence 5BP downstream 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: pJBL724
FEATURES             Location/Qualifiers
     source          1..2929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     120..137
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    254..275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(290..295,296..313,314..320)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    328..344
                     /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     333..355
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     352..368
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     352..368
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        411..429
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      609..655
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(681..698)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(681..697)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(690..712)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(906..925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1025..1047
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1085..1103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1171..1275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1276..1344,1345..2136)
                     /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(1494..1513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2307..2895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2796..2815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
       61 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
      121 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      181 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      241 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      301 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      361 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga taatacgact
      421 cactatagga ggtcagtagg tagacatcta agtatcccca catactctga tgatccgaga
      481 cggtcgggtc cagatattcg tatctgtcga gtagagtgtg ggctcggatc attcatggca
      541 agagacggtc gggtccagat attcgtatct gtcgagtaga gtgtgggctc ttgccatgtg
      601 tatgtgggta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgatccc
      661 cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
      721 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      781 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
      841 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
      901 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
      961 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     1021 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     1081 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1141 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1201 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1261 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1321 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1381 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1441 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1501 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1561 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1621 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1681 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1741 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1801 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1861 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1921 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1981 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2041 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2101 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2161 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2221 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctgg
//
