LOCUS       sequence-307666-        2919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TetR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the tetO sequence 0BP dowsntream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2919)
  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 2919)
  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: pJBL702
FEATURES             Location/Qualifiers
     source          1..2919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..19
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    136..157
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(172..177,178..195,196..202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    210..226
                     /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     215..237
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     234..250
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     234..250
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    310..328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      481..527
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(553..570)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(553..569)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(562..584)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(778..797)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     897..919
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(957..975)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1043..1147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1148..1216,1217..2008)
                     /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(1366..1385)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2179..2767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2668..2687
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
       61 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      121 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
      181 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      241 cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta gataatacga
      301 ctcactatat ccctatcagt gatagagacc cacatactct gatgatccga gacggtcggg
      361 tccagatatt cgtatctgtc gagtagagtg tgggctcgga tcattcatgg caagagacgg
      421 tcgggtccag atattcgtat ctgtcgagta gagtgtgggc tcttgccatg tgtatgtggg
      481 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgatc cccgggtacc
      541 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      601 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      661 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      721 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
      781 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      841 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      901 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
      961 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1021 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1081 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1141 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1201 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1261 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1321 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1381 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1441 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1501 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1561 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1621 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1681 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1741 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1801 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1861 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1921 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1981 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2041 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2101 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2161 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2221 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2281 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2341 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2401 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2461 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2521 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2581 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2641 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2701 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2761 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2821 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2881 gtgagctgat accgctcgcc gcagccgaac gaccgagcg
//
