LOCUS       sequence-133604-        4718 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. OG241 backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4718)
  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 4718)
  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-OG241
FEATURES             Location/Qualifiers
     source          1..4718
                     /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      complement(2314..2902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2394..2413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3648..4442
                     /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             3648..4442
                     /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(3702..3721)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4312..4331
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      4455..4591
                     /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 aatcatgtga gcaaaaggcc agcaaaaggc
     2281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3121 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     3181 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     3241 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     3301 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3361 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc
     3421 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     3481 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     3541 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     3601 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg attgaacaag
     3661 atggattgca cgcaggttct ccggcggctt gggtggagag gctattcggc tatgactggg
     3721 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgtc
     3781 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     3841 cgcggctatc gtggctggcg acgacgggcg ttccttgcgc ggctgtgctc gacgttgtca
     3901 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3961 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4021 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4081 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4141 tcgcgccagc cgaactgttc gccaggctca aggcgtctat gcccgacggc gaggatctcg
     4201 tcgtgaccca cggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4261 gattcatcga ctgtggccgt ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4321 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctt gtgctttacg
     4381 gtatcgccgc gcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4441 gaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     4501 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     4561 cgtgtcctcc ttaccagaaa tttatcctta aggtcgttta aactcgactc tggctctatc
     4621 gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat
     4681 cgaatctcgt cagctatcgt cagcttacct ttttggca
//
