LOCUS       sequence-297852-        2927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SmtB-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the smtO sequence 5BP downstream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2927)
  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 2927)
  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: pJBL726
FEATURES             Location/Qualifiers
     source          1..2927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     369..386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    503..524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(539..544,545..562,563..569)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    577..593
                     /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     582..604
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     601..617
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     601..617
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        660..678
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      856..902
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(928..945)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(928..944)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(937..959)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1153..1172)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1272..1294
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1332..1350)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1418..1522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1523..1591,1592..2383)
                     /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(1741..1760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2554..2927,1..215)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
       61 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      481 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      541 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      601 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagat
      661 aatacgactc actataggag gcacatgaac agttattcag atacccacat actctgatga
      721 tccgagacgg tcgggtccag atattcgtat ctgtcgagta gagtgtgggc tcggatcatt
      781 catggcaaga gacggtcggg tccagatatt cgtatctgtc gagtagagtg tgggctcttg
      841 ccatgtgtat gtgggtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      901 tgatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
      961 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1021 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1081 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     1141 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     1201 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     1261 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     1321 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     1381 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1441 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1501 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1561 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1621 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1681 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1741 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1801 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1861 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1921 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1981 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2041 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2101 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2161 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2221 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2281 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2341 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactga
//
