LOCUS       sequence-133603-        4719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Induced by acetoacetate to express GFP using atoSC two-component 
            system in E. coli. Contains the native atoDAEB promoter (Pato) from 
            E. coli DH5A. OG241 backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4719)
  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 4719)
  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: PatoG-OG241
FEATURES             Location/Qualifiers
     source          1..4719
                     /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             705..1415
                     /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    1525..1646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1536..1555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1590..1609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1761..1807
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1837..1973
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2018..2089
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2315..2903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2395..2414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3649..4443
                     /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             3649..4443
                     /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(3703..3722)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4313..4332
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      4456..4592
                     /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 tatgcaagag ggataaaaaa
      661 tgaaaacaaa attgatgaca ttacaagacg ccacggtacc caccatgaca gctctgacag
      721 agggagcgaa actgttcgag aaggagatcc cgtacatcac agagttggaa ggagatgtcg
      781 aagggatgaa attcatcatc aagggagaag gaacgggtga cgcaacgacc ggaacgatca
      841 aggccaaata catttgtacc acgggcgatt tgcctgtccc ctgggcgacg cttgtaagca
      901 cgctctcgta tggtgtccag tgcttcgcga aatatccatc gcacattaag gactttttca
      961 agtcggccat gccagaaggt tacacacaag aacgaaccat ctcctttgag ggggacggag
     1021 tgtataagac acgcgcgatg gtaacgtacg agcgcgggtc catctacaac agggtaactc
     1081 ttactgggga gaactttaag aaggacgggc atatcttgcg gaaaaacgtg gcatttcaat
     1141 gtccgccctc gattctgtat attctcccgg acacggtgaa taatgggatc agagtggagt
     1201 tcaaccaggc atacgatatt gagggtgtga ctgaaaagct cgtcaccaaa tgcagccaga
     1261 tgaatcgccc ccttgcggga tcagcagccg tccatatccc ccggtatcac catattacct
     1321 accacacaaa actctcaaaa gacagagatg agagaaggga ccatatgtgc ctggtcgaag
     1381 tagtgaaggc ggtggatctt gatacatacc agtgaaagtt gtctcctcct gcactgactg
     1441 actgatacaa tcgatttctg gatccgcagg cctctgctag cttgactgac tgagatacag
     1501 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     1561 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1621 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1681 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     1741 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1801 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     1861 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     1921 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     1981 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     2041 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     2101 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     2161 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     2221 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     2281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3121 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     3181 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     3241 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     3301 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     3361 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc
     3421 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     3481 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     3541 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     3601 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gattgaacaa
     3661 gatggattgc acgcaggttc tccggcggct tgggtggaga ggctattcgg ctatgactgg
     3721 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgt
     3781 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     3841 gcgcggctat cgtggctggc gacgacgggc gttccttgcg cggctgtgct cgacgttgtc
     3901 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3961 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4021 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4081 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4141 ctcgcgccag ccgaactgtt cgccaggctc aaggcgtcta tgcccgacgg cgaggatctc
     4201 gtcgtgaccc acggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4261 ggattcatcg actgtggccg tctgggtgtg gcggaccgct atcaggacat agcgttggct
     4321 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct tgtgctttac
     4381 ggtatcgccg cgcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4441 tgaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     4501 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     4561 acgtgtcctc cttaccagaa atttatcctt aaggtcgttt aaactcgact ctggctctat
     4621 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     4681 tcgaatctcg tcagctatcg tcagcttacc tttttggca
//
