LOCUS       sequence-307671-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 14BP dowsntream 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: pJBL710
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      234..822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     723..742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     976..993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1110..1131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1146..1151,1152..1169,1170..1176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1184..1200
                     /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     1189..1211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1208..1224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1208..1224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1267..1286
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1267..1285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1298..1316
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1469..1515
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1541..1558)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1541..1557)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1550..1572)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1766..1785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1885..1907
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1945..1963)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2031..2135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2136..2204,2205..2933,1..63)
                     /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(2354..2373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
       61 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      241 cctttttctc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
      421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      901 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      961 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     1021 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     1081 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     1141 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     1201 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac
     1261 tctagataat acgactcact atagggagac cacaacgtcc ctatcagtga tagagaccca
     1321 catactctga tgatccgaga cggtcgggtc cagatattcg tatctgtcga gtagagtgtg
     1381 ggctcggatc attcatggca agagacggtc gggtccagat attcgtatct gtcgagtaga
     1441 gtgtgggctc ttgccatgtg tatgtgggta gcataacccc ttggggcctc taaacgggtc
     1501 ttgaggggtt ttttgatccc cgggtaccga gctcgaattc actggccgtc gttttacaac
     1561 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1621 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1681 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     1741 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     1801 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     1861 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     1921 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     1981 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2041 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2101 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2161 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2221 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2281 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2341 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2401 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2461 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2521 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2581 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2641 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     2701 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2761 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2821 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2881 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gca
//
