LOCUS       sequence-133611-        6179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NOT induced by acetoacetate to express GFP. Contains the native 
            atoDAEB promoter (Pato) from E. coli DH5A, but out-of-frame with GFP
            gene. LOG241 backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  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 6179)
  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: PatoK-LOG241
FEATURES             Location/Qualifiers
     source          1..6179
                     /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             704..1414
                     /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    1524..1645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1535..1554)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1589..1608
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1760..1806
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1836..1972
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2017..2088
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      2733..2955
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5109..5903
                     /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             5109..5903
                     /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(5163..5182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5773..5792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5916..6052
                     /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 ccaggtaccc accatgacag ctctgacaga
      721 gggagcgaaa ctgttcgaga aggagatccc gtacatcaca gagttggaag gagatgtcga
      781 agggatgaaa ttcatcatca agggagaagg aacgggtgac gcaacgaccg gaacgatcaa
      841 ggccaaatac atttgtacca cgggcgattt gcctgtcccc tgggcgacgc ttgtaagcac
      901 gctctcgtat ggtgtccagt gcttcgcgaa atatccatcg cacattaagg actttttcaa
      961 gtcggccatg ccagaaggtt acacacaaga acgaaccatc tcctttgagg gggacggagt
     1021 gtataagaca cgcgcgatgg taacgtacga gcgcgggtcc atctacaaca gggtaactct
     1081 tactggggag aactttaaga aggacgggca tatcttgcgg aaaaacgtgg catttcaatg
     1141 tccgccctcg attctgtata ttctcccgga cacggtgaat aatgggatca gagtggagtt
     1201 caaccaggca tacgatattg agggtgtgac tgaaaagctc gtcaccaaat gcagccagat
     1261 gaatcgcccc cttgcgggat cagcagccgt ccatatcccc cggtatcacc atattaccta
     1321 ccacacaaaa ctctcaaaag acagagatga gagaagggac catatgtgcc tggtcgaagt
     1381 agtgaaggcg gtggatcttg atacatacca gtgaaagttg tctcctcctg cactgactga
     1441 ctgatacaat cgatttctgg atccgcaggc ctctgctagc ttgactgact gagatacagc
     1501 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1561 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1621 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1681 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1741 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1801 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     1861 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     1921 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     1981 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     2041 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     2101 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     2161 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     2221 cacccgttct ccgattacga gtttcattta aatgacagta agacgggtaa gcctgttgat
     2281 gataccgctg ccttactggg tgcattagcc agtctgaatg acctgtcacg ggataatccg
     2341 aagtggtcag actggaaaat cagagggcag gaactgctga acagcaaaaa gtcagatagc
     2401 accacatagc agacccgcca taaaacgccc tgagaagccc gtgacgggct tttcttgtat
     2461 tatgggtagt ttccttgcat gaatccataa aaggcgcctg tagtgccatt tacccccatt
     2521 cactgccaga gccgtgagcg cagcgaactg aatgtcacga aaaagacagc gactcaggtg
     2581 cctgatggtc ggagacaaaa ggaatattca gcgatttgcc cgagcttgcg agggtgctac
     2641 ttaagccttt agggttttaa ggtctgtttt gtagagcagc aaacagcgtt tgcgacatcc
     2701 ttttgtaata ctgcggaact gactaaagta gtgagttata cacagggctg ggatctattc
     2761 tttttatctt tttttattct ttctttattc tataaattat aaccacttga atataaacaa
     2821 aaaaaacaca caaaggtcta gcggaattta cagagggtct agcagaattt acaagttttc
     2881 cagcaaaggt ctagcagaat ttacagatac ccacaactca aaggaaaagg actagtaatt
     2941 atcattgact agcccatctc aattggtata gtgattaaaa tcacctagac caattgagat
     3001 gtatgtctga attagttgtt ttcaaagcaa atgaactagc gattagtcgc tatgacttaa
     3061 cggagcatga aaccaagcta attttatgct gtgtggcact actcaacccc acgattgaaa
     3121 accctacaag gaaagaacgg acggtatcgt ttcacttata accaatacgc tcagatgatg
     3181 aacatcagta gggaaaatgc ttatggtgta ttagctaaag caaccagaga gctgatgacg
     3241 agaactgtgg aaatcaggaa tcctttggtt aaaggctttg agattttcca gtggacaaac
     3301 tatgccaagt tctcaagcga aaaattagaa ttagttttta gtgaagagat attgccttat
     3361 cttttccagt taaaaaaatt cataaaatat aatctggaac atgttaagtc ttttgaaaac
     3421 aaatactcta tgaggattta tgagtggtta ttaaaagaac taacacaaaa gaaaactcac
     3481 aaggcaaata tagagattag ccttgatgaa tttaagttca tgttaatgct tgaaaataac
     3541 taccatgagt ttaaaaggct taaccaatgg gttttgaaac caataagtaa agatttaaac
     3601 acttacagca atatgaaatt ggtggttgat aagcgaggcc gcccgactga tacgttgatt
     3661 ttccaagttg aactagatag acaaatggat ctcgtaaccg aacttgagaa caaccagata
     3721 aaaatgaatg gtgacaaaat accaacaacc attacatcag attcctacct acgtaacgga
     3781 ctaagaaaaa cactacacga tgctttaact gcaaaaattc agctcaccag ttttgaggca
     3841 aaatttttga gtgacatgca aagtaagcat gatctcaatg gttcgttctc atggctcacg
     3901 caaaaacaac gaaccacact agagaacata ctggctaaat acggaaggat ctgaggttct
     3961 tatggctctt gtatctatca gtgaagcatc aagactaaca aacaaaagta gaacaactgt
     4021 tcaccgttac atatcaaagg gaaaactgtc catatgcaca gatgaaaacg gtgtaaaaaa
     4081 gatagataca tcagagcttt tacgagtttt tggtgcattt aaagctgttc accatgaaca
     4141 gatcgacaat gtaacagatg aacagcatgt aacacctaat agaacaggtg aaaccagtaa
     4201 aacaaagcaa ctagaacatg aaattgaaca cctgagacaa cttgttacag ctcaacagtc
     4261 acacatagac agcctgaaac aggcgatgct gcttatcgaa tcaaagctgc cgacaacacg
     4321 ggagccagtg acgcctcccg tggggaaaaa atcatggcaa ttctggaaga aatagcgctt
     4381 tcagccggca aacctgaagc cggatctgcg attctgataa caaactagca acaccagaac
     4441 agcccgtttg cgggcagcaa aacccgtact tttggacgtt ccggcggttt tttgtggcga
     4501 gtggtgttcg ggcggtgcgc gcaagatcca ttatgttaaa cgggcgagtt tacatctcaa
     4561 aaccgcccgc ttaacaccat cagaaatcct cagcgcgata tttaaatttc cgaactctcc
     4621 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     4681 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     4741 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     4801 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     4861 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     4921 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     4981 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     5041 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     5101 gctttcgcat gattgaacaa gatggattgc acgcaggttc tccggcggct tgggtggaga
     5161 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5221 ggctgtcagc gcaggggcgt ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5281 atgaactgca agacgaggca gcgcggctat cgtggctggc gacgacgggc gttccttgcg
     5341 cggctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5401 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5461 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5521 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5581 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgtcta
     5641 tgcccgacgg cgaggatctc gtcgtgaccc acggcgatgc ctgcttgccg aatatcatgg
     5701 tggaaaatgg ccgcttttct ggattcatcg actgtggccg tctgggtgtg gcggaccgct
     5761 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5821 accgcttcct tgtgctttac ggtatcgccg cgcccgattc gcagcgcatc gccttctatc
     5881 gccttcttga cgagttcttc tgaccgattc taggtgcatt ggcgcagaaa aaaatgcctg
     5941 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     6001 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgttt
     6061 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     6121 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggca
//
