LOCUS       sequence-384120-        5234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green Fluorescent Reporter of intracellular hydrogen peroxide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5234)
  AUTHORS   Kardashliev T, Weingartner A, Romero E, Schwaneberg U, Fraaije M, 
            Panke S, Held M
  TITLE     Whole-cell screening of oxidative enzymes using genetically encoded 
            sensors.
  JOURNAL   Chem Sci. 2021 Oct 29;12(44):14766-14772. doi: 10.1039/d1sc02578c. 
            eCollection 2021 Nov 17.
  PUBMED    34820092
REFERENCE   2  (bases 1 to 5234)
  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: 
            "10.1039/d1sc02578c"; journalName: "Chem Sci"; date: "2021-10-29- 
            29"; volume: "12"; issue: "44"; pages: "14766-14772"
COMMENT     Sequence Label: pOxyR_gfp
FEATURES             Location/Qualifiers
     source          1..5234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            205..313
                     /note="incP origin of transfer"
     CDS             complement(728..1678)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     WTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1726..1948
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2281..2367)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(2391..3101,3102..3104))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(2391..3104)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2431..2451)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3046..3074
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      4172..4266
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             join(4502..5234,1..59)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataag
       61 acaattgtcc ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
      121 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
      181 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
      241 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
      301 gcgaggctgg ccgtaggccg gccgccatga ttttttcccc acgggaggcg tcactggctc
      361 ccgtgttgtc ggcagctttg attcgataag cagcatcgcc tgtttcaggc tgtctatgtg
      421 tgactgttga gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta gttgctttgt
      481 tttactggtt tcacctgttc tattaggtgt tacatgctgt tcatctgtta cattgtcgat
      541 ctgttcatgg tgaacagctt taaatgcacc aaaaactcgt aaaagctctg atgtatctat
      601 cttttttaca ccgttttcat ctgtgcatat ggacagtttt ccctttgata tgtaacggtg
      661 aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata caagagccat
      721 aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt cgttgttttt
      781 gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca ctcaaaaatt
      841 ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt gtttttctta
      901 gtccgttatg taggtaggaa tctgatgtaa tggttgttgg tattttgtcg ccattcattt
      961 ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt tcaacttgga
     1021 aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata ttgctgtaag
     1081 tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta aactcatggt
     1141 agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct atatttgcct
     1201 tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc atagagtatt
     1261 tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt aactggaaaa
     1321 gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag aacttggcat
     1381 agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt tccacagttc
     1441 tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc ctactgatgt
     1501 tcatcatctg agcgtattgg ttataagtga acgataccgt ccattctttc cttgtagggt
     1561 tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt tcatgctccg
     1621 ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat tcagacatac
     1681 atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct agtcaatgat
     1741 aattactagt ccttttcctt tgagttgtgg gtatctgtaa attctgctag acctttgctg
     1801 gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg tgtgtttttt
     1861 ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa aagataaaaa
     1921 gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag tattacaaaa
     1981 ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct aaaggcttaa
     2041 gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg
     2101 cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg
     2161 aatgggggta aatggcacta caggcgccga ataaatacct gtgacggaag atcacttcgc
     2221 agaataaata aatcctggtg tccctgttga tacggcgcgc ccagctgtct agggcggcgg
     2281 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     2341 tttcgactga gcctttcgtt ttatttgatg cctttaatta attaggatcc tttgtagagc
     2401 tcatccatgc catgtgtaat cccagcagca gttacaaact caagaaggac catgtggtca
     2461 cgcttttcgt tgggatcttt cgaaaggaca gattgtgtcg acaggtaatg gttgtctggt
     2521 aaaaggacag ggccatcgcc aattggagta ttttgttgat aatggtctgc tagttgaacg
     2581 gaaccatctt caacgttgtg gcgaattttg aagttagctt tgattccatt cttttgtttg
     2641 tctgccgtga tgtatacatt gtgtgagtta aagttgtact cgagtttgtg tccgagaatg
     2701 tttccatctt ctttaaaatc aatacctttt aactcgatac gattaacaag ggtatcacct
     2761 tcaaacttga cttcagcacg cgtcttgtag gtcccgtcat ctttgaaaga tatagtgcgt
     2821 tcctgtacat aaccttcggg catggcactc ttgaaaaagt catgccgttt catgtgatcc
     2881 ggataacggg aaaagcattg aacaccatag gtcagagtag tgacaagtgt tggccatgga
     2941 acaggtagtt ttccagtagt gcaaataaat ttaagggtga gttttccgtt tgtagcatca
     3001 ccttcaccct ctcctctgac agaaaatttg tgcccattaa catcaccatc taattcaaca
     3061 agaattggga caactccagt gaaaagttct tctcctttgc tcatatgtta aatctcctgc
     3121 taattaaaaa gtaaagtgtg aattctgctg tgagcaatta tcagtcagaa tgcttgatag
     3181 ggataatcgt tcattgctat tctacctatc gccatgaact atcgtggcga tggaggatgg
     3241 ataatgaata ttcgtgatct tgagtacctg gtggcattgg ctgaacaccg ccattttcgg
     3301 cgtgcggcag attcctgcca cgttagccag ccgacgctta gcgggcaaat tcgtaagctg
     3361 gaagatgagc tgggcgtgat gttgctggag cggaccagcc gtaaagtgtt gttcacccag
     3421 gcgggaatgc tgctggtgga tcaggcgcgt accgtgctgc gtgaggtgaa agtccttaaa
     3481 gagatggcaa gccagcaggg cgagacgatg tccggaccgc tgcacattgg tttgattccc
     3541 acagttggac cgtacctgct accgcatatt atccctatgc tgcaccagac ctttccaaag
     3601 ctggaaatgt atctgcatga agcacagacc caccagttac tggcgcaact ggacagcggc
     3661 aaactcgatt gcgtgatcct cgcgctggtg aaagagagcg aagcattcat tgaagtgccg
     3721 ttgtttgatg agccaatgtt gctggctatc tatgaagatc acccgtgggc gaaccgcgaa
     3781 tgcgtaccga tggccgatct ggcaggggaa aaactgctga tgctggaaga tggtcactgt
     3841 ttgcgcgatc aggcaatggg tttctgtttt gaagccgggg cggatgaaga tacacacttc
     3901 cgcgcgacca gcctggaaac tctgcgcaac atggtggcgg caggtagcgg gatcacttta
     3961 ctgccagcgc tggctgtgcc gccggagcgc aaacgcgatg gggttgttta tctgccgtgc
     4021 attaagccgg aaccacgccg cactattggc ctggtttatc gtcctggctc accgctgcgc
     4081 agccgctatg agcagctggc agaggccatc cgcgcaagaa tggatggcca tttcgataaa
     4141 gttttaaaac aggcggttta aactagtctt ggactcctgt tgatagatcc agtaatgacc
     4201 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     4261 gagaatccag gggtccccaa taattacgat ttacgtattt aaatgaacct tgaccgaacg
     4321 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     4381 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     4441 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     4501 catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     4561 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     4621 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     4681 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     4741 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     4801 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     4861 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     4921 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     4981 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     5041 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     5101 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     5161 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     5221 ctcgcgcgca gatc
//
