LOCUS       sequence-311292-        4220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression (sigma 70 promoter) of the ratiometric 
            mRuby-iATPSnFR1.0 ATP indicator in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4220)
  AUTHORS   Bruni GN, Kralj JM
  TITLE     Membrane voltage dysregulation driven by metabolic dysfunction 
            underlies bactericidal activity of aminoglycosides.
  JOURNAL   eLife. 2020 Aug 4;9. pii: 58706. doi: 10.7554/eLife.58706.
  PUBMED    32748785
REFERENCE   2  (bases 1 to 4220)
  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: "eLife."; 
            date: "2020-08-4"; volume: "9. pii: 58706. doi"; pages: " 
            10.7554/eLife.58706"
COMMENT     Sequence Label: pKL10-mRuby-iATPSnFR1.0 bb118
FEATURES             Location/Qualifiers
     source          1..4220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(48..636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(128..147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(807..1598,1599..1667))
                     /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     1430..1449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1668..1772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1840..1858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1862..1905)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1908..1929
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        1930..1964
                     /label=BBa_J23118
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.56"
     primer_bind     complement(3883..3900)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     primer_bind     complement(4097..4114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccacaggctc
       61 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     1801 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag
     1861 aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg
     1921 cttctagagt tgacggctag ctcagtccta ggtattgtgc tagcattaac tttaagaagg
     1981 agatatacat acccatggtc tccaaggggg aggaacttat taaagaaaat atgcgcatga
     2041 aagttgttat ggaggggtct gtaaatggtc accagtttaa gtgcacaggg gagggcgagg
     2101 gacgtcctta tgaaggggtc caaaccatgc gcattaaggt gattgagggt ggcccgcttc
     2161 cgttcgcttt tgacatcctt gctacaagtt ttatgtatgg aagtcgtacc tttatcaaat
     2221 atcccgcaga tattccagat tttttcaagc agagtttccc tgagggcttc acatgggagc
     2281 gtgtcacccg ttacgaagac ggtggtgtag tcaccgtcac ccaagataca tctctggaag
     2341 acggtgaact ggtgtacaac gtaaaagtcc gtggcgtaaa ttttccatct aacggaccgg
     2401 tcatgcagaa gaaaaccaaa ggttgggaac ctaacacaga gatgatgtat cccgcagacg
     2461 ggggcttacg tggctatacg gacatcgctt taaaggtgga cggagggggt caccttcact
     2521 gtaattttgt cacgacgtat cgttccaaga aaacagttgg aaacatcaag atgcctgggg
     2581 tacatgcagt agaccatcgc cttgaacgca tcgaggagtc cgataatgag acttacgtcg
     2641 tgcagcgtga ggtagcagtc gccaaatact cgaacctggg cgggggctcg atgaagacaa
     2701 ttcatgtctc tgtagttaca cctgatggac ccgtgtacga ggacgacgtc gagatggttt
     2761 ccgtcaaggc aaaatctggt gagctgggca tcttaccagg acatatccca ctgaaggccc
     2821 ccctggaaat ctccgcggcc cgcttaaaaa agggaggtaa gacacaatac attgcagtga
     2881 gcgggggtaa tctggaggtt cgccctgaca aggtcacaat ctatgctcag gcagcagagc
     2941 gcgcggaaga catcgatgta ctgcgtgcca aagcagccaa ggaacgtgca gaacgtcgcc
     3001 ttcagagcca agtgctgtct cacaacgtgt acattacggc tgacaaacaa aaaaacggga
     3061 tcaaagccaa tttcaaaatt cgccataatg tggaggacgg cagcatgcaa ttggctgacc
     3121 attatcagca aaataccccg atcggagatg gcccggtgct tcttccagat aaccattacc
     3181 ttagtacgca gtccgtcctg tcaaaggatc cgaatgagaa gcgtgatcat atggtgttac
     3241 tggagtttgt gactgctgct ggcattacgc ttgggatgga cgagctttac aagggaggta
     3301 ccggggggtc tatgtcgaag ggtgaagaat tgtttaccgg agtagttccg attcttgtag
     3361 agctggatgg tgatgttaac gggcataagt tctcagtgcg tggtgaagga gagggggatg
     3421 ccactaatgg gaagttgact cttaaattta tctgcacaac tggaaaactt cctgttccct
     3481 ggccaacctt agtcaccacc ttaacgtacg gagttcagtg tttcagccgc tatccagacc
     3541 acatgaagca acacgatttc tttaaatcgg ccatgcctga gggttacgtt caggaacgta
     3601 cgattagttt taaggatgac ggcacctata agacgcgtgc tgaagttaag tttgaaggtg
     3661 atactctggt aaaccgcatt gaattaaagg gaatcgattt caaagaagac ggtaacatct
     3721 taggacataa gttagagtat aacttcgggt tgcatgatat cgacttcaaa cgtgctgaat
     3781 tagcccttaa acgtgcgatg aaccgtctgt ctgtcgcgga gatgaagtag gtttaaacgg
     3841 tctccagctt ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
     3901 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     3961 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     4021 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     4081 actgggcctt ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4141 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4201 gaaagaacat gtgagcaaaa
//
