LOCUS       sequence-133612-        6156 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, Bba_B0034 (iGEM). LOG241 backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6156)
  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 6156)
  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: PatoL-LOG241
FEATURES             Location/Qualifiers
     source          1..6156
                     /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"
     RBS             651..662
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             681..1391
                     /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    1501..1622
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1512..1531)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1566..1585
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1737..1783
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1813..1949
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1994..2065
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      2710..2932
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5086..5880
                     /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             5086..5880
                     /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(5140..5159)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5750..5769
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5893..6029
                     /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 tatgcagatt aaagaggaga
      661 aatactagat ggtacccacc atgacagctc tgacagaggg agcgaaactg ttcgagaagg
      721 agatcccgta catcacagag ttggaaggag atgtcgaagg gatgaaattc atcatcaagg
      781 gagaaggaac gggtgacgca acgaccggaa cgatcaaggc caaatacatt tgtaccacgg
      841 gcgatttgcc tgtcccctgg gcgacgcttg taagcacgct ctcgtatggt gtccagtgct
      901 tcgcgaaata tccatcgcac attaaggact ttttcaagtc ggccatgcca gaaggttaca
      961 cacaagaacg aaccatctcc tttgaggggg acggagtgta taagacacgc gcgatggtaa
     1021 cgtacgagcg cgggtccatc tacaacaggg taactcttac tggggagaac tttaagaagg
     1081 acgggcatat cttgcggaaa aacgtggcat ttcaatgtcc gccctcgatt ctgtatattc
     1141 tcccggacac ggtgaataat gggatcagag tggagttcaa ccaggcatac gatattgagg
     1201 gtgtgactga aaagctcgtc accaaatgca gccagatgaa tcgccccctt gcgggatcag
     1261 cagccgtcca tatcccccgg tatcaccata ttacctacca cacaaaactc tcaaaagaca
     1321 gagatgagag aagggaccat atgtgcctgg tcgaagtagt gaaggcggtg gatcttgata
     1381 cataccagtg aaagttgtct cctcctgcac tgactgactg atacaatcga tttctggatc
     1441 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     1501 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1561 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     1621 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     1681 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     1741 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     1801 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     1861 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     1921 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
     1981 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
     2041 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
     2101 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     2161 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     2221 tcatttaaat gacagtaaga cgggtaagcc tgttgatgat accgctgcct tactgggtgc
     2281 attagccagt ctgaatgacc tgtcacggga taatccgaag tggtcagact ggaaaatcag
     2341 agggcaggaa ctgctgaaca gcaaaaagtc agatagcacc acatagcaga cccgccataa
     2401 aacgccctga gaagcccgtg acgggctttt cttgtattat gggtagtttc cttgcatgaa
     2461 tccataaaag gcgcctgtag tgccatttac ccccattcac tgccagagcc gtgagcgcag
     2521 cgaactgaat gtcacgaaaa agacagcgac tcaggtgcct gatggtcgga gacaaaagga
     2581 atattcagcg atttgcccga gcttgcgagg gtgctactta agcctttagg gttttaaggt
     2641 ctgttttgta gagcagcaaa cagcgtttgc gacatccttt tgtaatactg cggaactgac
     2701 taaagtagtg agttatacac agggctggga tctattcttt ttatcttttt ttattctttc
     2761 tttattctat aaattataac cacttgaata taaacaaaaa aaacacacaa aggtctagcg
     2821 gaatttacag agggtctagc agaatttaca agttttccag caaaggtcta gcagaattta
     2881 cagataccca caactcaaag gaaaaggact agtaattatc attgactagc ccatctcaat
     2941 tggtatagtg attaaaatca cctagaccaa ttgagatgta tgtctgaatt agttgttttc
     3001 aaagcaaatg aactagcgat tagtcgctat gacttaacgg agcatgaaac caagctaatt
     3061 ttatgctgtg tggcactact caaccccacg attgaaaacc ctacaaggaa agaacggacg
     3121 gtatcgtttc acttataacc aatacgctca gatgatgaac atcagtaggg aaaatgctta
     3181 tggtgtatta gctaaagcaa ccagagagct gatgacgaga actgtggaaa tcaggaatcc
     3241 tttggttaaa ggctttgaga ttttccagtg gacaaactat gccaagttct caagcgaaaa
     3301 attagaatta gtttttagtg aagagatatt gccttatctt ttccagttaa aaaaattcat
     3361 aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa tactctatga ggatttatga
     3421 gtggttatta aaagaactaa cacaaaagaa aactcacaag gcaaatatag agattagcct
     3481 tgatgaattt aagttcatgt taatgcttga aaataactac catgagttta aaaggcttaa
     3541 ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact tacagcaata tgaaattggt
     3601 ggttgataag cgaggccgcc cgactgatac gttgattttc caagttgaac tagatagaca
     3661 aatggatctc gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg acaaaatacc
     3721 aacaaccatt acatcagatt cctacctacg taacggacta agaaaaacac tacacgatgc
     3781 tttaactgca aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag
     3841 taagcatgat ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa ccacactaga
     3901 gaacatactg gctaaatacg gaaggatctg aggttcttat ggctcttgta tctatcagtg
     3961 aagcatcaag actaacaaac aaaagtagaa caactgttca ccgttacata tcaaagggaa
     4021 aactgtccat atgcacagat gaaaacggtg taaaaaagat agatacatca gagcttttac
     4081 gagtttttgg tgcatttaaa gctgttcacc atgaacagat cgacaatgta acagatgaac
     4141 agcatgtaac acctaataga acaggtgaaa ccagtaaaac aaagcaacta gaacatgaaa
     4201 ttgaacacct gagacaactt gttacagctc aacagtcaca catagacagc ctgaaacagg
     4261 cgatgctgct tatcgaatca aagctgccga caacacggga gccagtgacg cctcccgtgg
     4321 ggaaaaaatc atggcaattc tggaagaaat agcgctttca gccggcaaac ctgaagccgg
     4381 atctgcgatt ctgataacaa actagcaaca ccagaacagc ccgtttgcgg gcagcaaaac
     4441 ccgtactttt ggacgttccg gcggtttttt gtggcgagtg gtgttcgggc ggtgcgcgca
     4501 agatccatta tgttaaacgg gcgagtttac atctcaaaac cgcccgctta acaccatcag
     4561 aaatcctcag cgcgatattt aaatttccga actctccaag gccctcgtcg gaaaatcttc
     4621 aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt
     4681 ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc
     4741 ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc
     4801 tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc
     4861 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     4921 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     4981 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     5041 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgat tgaacaagat
     5101 ggattgcacg caggttctcc ggcggcttgg gtggagaggc tattcggcta tgactgggca
     5161 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgtccg
     5221 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5281 cggctatcgt ggctggcgac gacgggcgtt ccttgcgcgg ctgtgctcga cgttgtcact
     5341 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5401 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5461 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5521 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5581 gcgccagccg aactgttcgc caggctcaag gcgtctatgc ccgacggcga ggatctcgtc
     5641 gtgacccacg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5701 ttcatcgact gtggccgtct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5761 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttccttgt gctttacggt
     5821 atcgccgcgc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5881 ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     5941 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     6001 tgtcctcctt accagaaatt tatccttaag gtcgtttaaa ctcgactctg gctctatcga
     6061 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     6121 aatctcgtca gctatcgtca gcttaccttt ttggca
//
