LOCUS       sequence-297843-        2936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MphR-regulated 3WJdB RNA aptamer driven by T7 promoter 
            with the mphO sequence 5BP downstream from the promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2936)
  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 2936)
  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: pJBL716
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..76
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    193..214
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(229..234,235..252,253..259)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    267..283
                     /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     272..294
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     291..307
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     291..307
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        350..368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      558..604
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(627..644)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(627..643)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(636..658)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(852..871)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     971..993
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1031..1049)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1117..1221
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1222..1290,1291..2082)
                     /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(1440..1459)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2253..2841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2742..2761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
       61 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
      121 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
      181 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
      241 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      301 ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagat aatacgactc
      361 actataggag ggaatataac cgacgtgact gttacattta ggtggcccac atactctgat
      421 gatccgagac ggtcgggtcc agatattcgt atctgtcgag tagagtgtgg gctcggatca
      481 ttcatggcaa gagacggtcg ggtccagata ttcgtatctg tcgagtagag tgtgggctct
      541 tgccatgtgt atgtgggtag cataacccct tggggcctct aaacgggtct tgaggggttt
      601 tttgcccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
      661 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      721 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      781 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
      841 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
      901 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      961 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     1021 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1081 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1141 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1201 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1261 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1321 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1381 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1441 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1501 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1561 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1621 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1681 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1741 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     1801 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     1861 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     1921 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     1981 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2041 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2101 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2161 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2221 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2281 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2341 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2401 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2461 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2521 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2581 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2641 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2701 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2761 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2821 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2881 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataa
//
