LOCUS       sequence-133606-        4683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Induced by acetoacetate to express GFP using atoSC two-component 
            system in E. coli. Contains the atoDAEB promoter (Pato) with a 
            strong RBS, Shine-Dalgarno sequence provided in OG241. OG241 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4683)
  AUTHORS   Rutter JW, Dekker L, Fedorec AJH, Gonzales DT, Wen KY, Tanner LES, 
            Donovan E, Ozdemir T, Thomas GM, Barnes CP
  TITLE     Engineered acetoacetate-inducible whole-cell biosensors based on the
            AtoSC two-component system.
  JOURNAL   Biotechnol Bioeng. 2021 Nov;118(11):4278-4289. doi: 
            10.1002/bit.27897. Epub 2021 Aug 9.
  PUBMED    34289076
REFERENCE   2  (bases 1 to 4683)
  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"; doi: "10.1002/bit.27897";
            journalName: "Biotechnol Bioeng"; date: "2021-11"; volume: "118"; 
            issue: "11"; pages: "4278-4289"
COMMENT     Sequence Label: PatoM-OG241
FEATURES             Location/Qualifiers
     source          1..4683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..89
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(94..140)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    158..206
                     /note="synthetic polyadenylation signal"
     polyA_signal    241..289
                     /note="synthetic polyadenylation signal"
     misc_feature    303..394
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     343..362
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             669..1379
                     /codon_start=1
                     /product="green fluorescent protein with peak excitation at
                     510 nm and peak emission at 521 nm"
                     /label=daGFP
                     /translation="MTALTEGAKLFEKEIPYITELEGDVEGMKFIIKGEGTGDATTGTI
                     KAKYICTTGDLPVPWATLVSTLSYGVQCFAKYPSHIKDFFKSAMPEGYTQERTISFEGD
                     GVYKTRAMVTYERGSIYNRVTLTGENFKKDGHILRKNVAFQCPPSILYILPDTVNNGIR
                     VEFNQAYDIEGVTEKLVTKCSQMNRPLAGSAAVHIPRYHHITYHTKLSKDRDERRDHMC
                     LVEVVKAVDLDTYQ"
     polyA_signal    1489..1610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1500..1519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1554..1573
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1725..1771
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1801..1937
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1982..2053
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2279..2867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2359..2378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3613..4407
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3613..4407
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3667..3686)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4277..4296
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      4420..4556
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
       61 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
      121 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
      181 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctgat
      241 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgaaagta
      301 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      361 cccagtgcaa atgcaggtgc cagaacattt ctctgtcgac caactagtgc cgtgcattga
      421 tgtataaact ccaggaatac ggtatcgatc cggcggatgt ataacaccaa aacttgctat
      481 gcagaaattt gcacagtgcg caattttctg catagccgct cattctcctt ataaatcccc
      541 atccaattta tcccttcata ttcaattagt taaataacta aatccaataa tctcattctg
      601 gcactcccct tgctattgcc tgactgtacc cacaacggtg tatgcagaat tcaaaggagg
      661 tacccaccat gacagctctg acagagggag cgaaactgtt cgagaaggag atcccgtaca
      721 tcacagagtt ggaaggagat gtcgaaggga tgaaattcat catcaaggga gaaggaacgg
      781 gtgacgcaac gaccggaacg atcaaggcca aatacatttg taccacgggc gatttgcctg
      841 tcccctgggc gacgcttgta agcacgctct cgtatggtgt ccagtgcttc gcgaaatatc
      901 catcgcacat taaggacttt ttcaagtcgg ccatgccaga aggttacaca caagaacgaa
      961 ccatctcctt tgagggggac ggagtgtata agacacgcgc gatggtaacg tacgagcgcg
     1021 ggtccatcta caacagggta actcttactg gggagaactt taagaaggac gggcatatct
     1081 tgcggaaaaa cgtggcattt caatgtccgc cctcgattct gtatattctc ccggacacgg
     1141 tgaataatgg gatcagagtg gagttcaacc aggcatacga tattgagggt gtgactgaaa
     1201 agctcgtcac caaatgcagc cagatgaatc gcccccttgc gggatcagca gccgtccata
     1261 tcccccggta tcaccatatt acctaccaca caaaactctc aaaagacaga gatgagagaa
     1321 gggaccatat gtgcctggtc gaagtagtga aggcggtgga tcttgataca taccagtgaa
     1381 agttgtctcc tcctgcactg actgactgat acaatcgatt tctggatccg caggcctctg
     1441 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     1501 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1561 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1621 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1681 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     1741 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     1801 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     1861 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     1921 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     1981 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     2041 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     2101 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     2161 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     2221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2461 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2701 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2941 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3001 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3061 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     3121 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     3181 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     3241 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     3301 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     3361 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
     3421 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
     3481 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
     3541 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
     3601 ggaggctttc gcatgattga acaagatgga ttgcacgcag gttctccggc ggcttgggtg
     3661 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3721 ttccggctgt cagcgcaggg gcgtccggtt ctttttgtca agaccgacct gtccggtgcc
     3781 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggcgacgac gggcgttcct
     3841 tgcgcggctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3901 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3961 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4021 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4081 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4141 tctatgcccg acggcgagga tctcgtcgtg acccacggcg atgcctgctt gccgaatatc
     4201 atggtggaaa atggccgctt ttctggattc atcgactgtg gccgtctggg tgtggcggac
     4261 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4321 gctgaccgct tccttgtgct ttacggtatc gccgcgcccg attcgcagcg catcgccttc
     4381 tatcgccttc ttgacgagtt cttctgaccg attctaggtg cattggcgca gaaaaaaatg
     4441 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     4501 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
     4561 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     4621 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     4681 gca
//
