LOCUS       sequence-133613-        6144 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. LOG241 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6144)
  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 6144)
  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-LOG241
FEATURES             Location/Qualifiers
     source          1..6144
                     /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      2698..2920
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5074..5868
                     /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             5074..5868
                     /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(5128..5147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5738..5757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5881..6017
                     /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 atttaaatga
     2221 cagtaagacg ggtaagcctg ttgatgatac cgctgcctta ctgggtgcat tagccagtct
     2281 gaatgacctg tcacgggata atccgaagtg gtcagactgg aaaatcagag ggcaggaact
     2341 gctgaacagc aaaaagtcag atagcaccac atagcagacc cgccataaaa cgccctgaga
     2401 agcccgtgac gggcttttct tgtattatgg gtagtttcct tgcatgaatc cataaaaggc
     2461 gcctgtagtg ccatttaccc ccattcactg ccagagccgt gagcgcagcg aactgaatgt
     2521 cacgaaaaag acagcgactc aggtgcctga tggtcggaga caaaaggaat attcagcgat
     2581 ttgcccgagc ttgcgagggt gctacttaag cctttagggt tttaaggtct gttttgtaga
     2641 gcagcaaaca gcgtttgcga catccttttg taatactgcg gaactgacta aagtagtgag
     2701 ttatacacag ggctgggatc tattcttttt atcttttttt attctttctt tattctataa
     2761 attataacca cttgaatata aacaaaaaaa acacacaaag gtctagcgga atttacagag
     2821 ggtctagcag aatttacaag ttttccagca aaggtctagc agaatttaca gatacccaca
     2881 actcaaagga aaaggactag taattatcat tgactagccc atctcaattg gtatagtgat
     2941 taaaatcacc tagaccaatt gagatgtatg tctgaattag ttgttttcaa agcaaatgaa
     3001 ctagcgatta gtcgctatga cttaacggag catgaaacca agctaatttt atgctgtgtg
     3061 gcactactca accccacgat tgaaaaccct acaaggaaag aacggacggt atcgtttcac
     3121 ttataaccaa tacgctcaga tgatgaacat cagtagggaa aatgcttatg gtgtattagc
     3181 taaagcaacc agagagctga tgacgagaac tgtggaaatc aggaatcctt tggttaaagg
     3241 ctttgagatt ttccagtgga caaactatgc caagttctca agcgaaaaat tagaattagt
     3301 ttttagtgaa gagatattgc cttatctttt ccagttaaaa aaattcataa aatataatct
     3361 ggaacatgtt aagtcttttg aaaacaaata ctctatgagg atttatgagt ggttattaaa
     3421 agaactaaca caaaagaaaa ctcacaaggc aaatatagag attagccttg atgaatttaa
     3481 gttcatgtta atgcttgaaa ataactacca tgagtttaaa aggcttaacc aatgggtttt
     3541 gaaaccaata agtaaagatt taaacactta cagcaatatg aaattggtgg ttgataagcg
     3601 aggccgcccg actgatacgt tgattttcca agttgaacta gatagacaaa tggatctcgt
     3661 aaccgaactt gagaacaacc agataaaaat gaatggtgac aaaataccaa caaccattac
     3721 atcagattcc tacctacgta acggactaag aaaaacacta cacgatgctt taactgcaaa
     3781 aattcagctc accagttttg aggcaaaatt tttgagtgac atgcaaagta agcatgatct
     3841 caatggttcg ttctcatggc tcacgcaaaa acaacgaacc acactagaga acatactggc
     3901 taaatacgga aggatctgag gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac
     3961 taacaaacaa aagtagaaca actgttcacc gttacatatc aaagggaaaa ctgtccatat
     4021 gcacagatga aaacggtgta aaaaagatag atacatcaga gcttttacga gtttttggtg
     4081 catttaaagc tgttcaccat gaacagatcg acaatgtaac agatgaacag catgtaacac
     4141 ctaatagaac aggtgaaacc agtaaaacaa agcaactaga acatgaaatt gaacacctga
     4201 gacaacttgt tacagctcaa cagtcacaca tagacagcct gaaacaggcg atgctgctta
     4261 tcgaatcaaa gctgccgaca acacgggagc cagtgacgcc tcccgtgggg aaaaaatcat
     4321 ggcaattctg gaagaaatag cgctttcagc cggcaaacct gaagccggat ctgcgattct
     4381 gataacaaac tagcaacacc agaacagccc gtttgcgggc agcaaaaccc gtacttttgg
     4441 acgttccggc ggttttttgt ggcgagtggt gttcgggcgg tgcgcgcaag atccattatg
     4501 ttaaacgggc gagtttacat ctcaaaaccg cccgcttaac accatcagaa atcctcagcg
     4561 cgatatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     4621 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     4681 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     4741 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     4801 gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc
     4861 tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga
     4921 agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa
     4981 cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc
     5041 tatcaagaga caggatcagc aggaggcttt cgcatgattg aacaagatgg attgcacgca
     5101 ggttctccgg cggcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5161 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgtccggt tctttttgtc
     5221 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     5281 ctggcgacga cgggcgttcc ttgcgcggct gtgctcgacg ttgtcactga agcgggaagg
     5341 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5401 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5461 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5521 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5581 ctgttcgcca ggctcaaggc gtctatgccc gacggcgagg atctcgtcgt gacccacggc
     5641 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5701 ggccgtctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5761 gaagagcttg gcggcgaatg ggctgaccgc ttccttgtgc tttacggtat cgccgcgccc
     5821 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgacc gattctaggt
     5881 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     5941 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     6001 cagaaattta tccttaaggt cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     6061 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     6121 tatcgtcagc ttaccttttt ggca
//
