LOCUS       Exported                5531 bp DNA     circular SYN 31-JUL-2024
DEFINITION  Mammalian expression of non-targeted ultrasensitive hydrogen 
            peroxide indicator HyPer7 for optical imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5531)
  AUTHORS   Pak VV, Ezerina D, Lyublinskaya OG, Pedre B, Tyurin-Kuzmin PA, 
            Mishina NM, Thauvin M, Young D, Wahni K, Martinez Gache SA, 
            Demidovich AD, Ermakova YG, Maslova YD, Shokhina AG, Eroglu E, Bilan
            DS, Bogeski I, Michel T, Vriz S, Messens J, Belousov VV
  TITLE     Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide 
            Identifies Roles for the Oxidant in Cell Migration and Mitochondrial
            Function.
  JOURNAL   Cell Metab. 2020 Mar 3;31(3):642-653.e6. doi: 
            10.1016/j.cmet.2020.02.003.
  PUBMED    32130885
REFERENCE   2  (bases 1 to 5531)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5531)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5531)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cmet.2020.02"; journalName: "Cell Metab"; date: 
            "2020-03-3- 3"; volume: "31"; issue: "3"; pages: "642-653.e6"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             106..489
                     /codon_start=1
                     /product="OxyR-RD domain from Neisseria meningitidis"
                     /label=oxyR
                     /translation="MHLANEEQNELEGAFKLGLIFTVAPYLLPKLIVSLRRTAPKMPLM
                     LEENYTHTLTESLKRGDVDAIIVAEPFQEPGIVTEPLYDEPFFVIVPKGHSFEELDAVS
                     PRMLGEEQVLLLTEGNCMRDQVLS"
     CDS             511..1236
                     /codon_start=1
                     /label=cpGFP
                     /note="circularly permuted GFP"
                     /translation="NVYIMADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPV
                     LLPDNHYLSFQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDELYNVDGGSGGTGSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLS
                     YGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIGFKEDGNILGHKLEYN"
     primer_bind     717..738
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(847..868)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1108..1127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1243..1542
                     /codon_start=1
                     /product="OxyR-RD domain from Neisseria meningitidis"
                     /label=oxyR
                     /translation="SCSELAAKQRIQGLTNTLQGSSINTIRHMVASGLAISVLPATALT
                     ENDHMLFSIIPFEGTPPSRRVVLAYRRNFVRPKALSAMKAAIMQSQLHGVSFICD"
     promoter        complement(1561..1578)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    complement(1583..1717)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1830..1848)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1869..1885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1893..1909)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(1917..1947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1962..1983)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(2100..2117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2271..2859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3033..3890)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3891..3995)
                     /label=AmpR promoter
     rep_origin      complement(4021..4476)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4553..5531
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg ccaccatgca cctggctaat
      121 gaggagcaaa acgaattgga aggtgctttt aagctgggct tgatctttac cgtcgccccg
      181 tacttgctgc ctaagttgat cgtcagcctg cgccgcaccg ccccgaagat gcctttgatg
      241 ttggaggaga attacaccca cacccttacc gagtcactga aacgcggcga cgttgacgcc
      301 atcattgtgg cagagccgtt tcaagagccg ggtattgtga ctgaaccttt gtatgacgag
      361 ccgtttttcg ttatcgttcc gaagggccat agcttcgagg agttggatgc cgttagccct
      421 cgcatgttgg gcgaagaaca agttctgctt ctgaccgagg gcaactgtat gcgtgaccag
      481 gtgcttagct ccgccggctt caacagcaac aacgtctata tcatggccga caagcagaag
      541 aacggcatca aggccaactt caagatccgc cacaacgtcg aggacggcag cgtgcagctc
      601 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      661 cactacctga gcttccagtc cgtcctgagc aaagacccca acgagaagcg cgatcacatg
      721 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaac
      781 gtggatggcg gtagcggtgg caccggcagc aagggcgagg agctgttcac cggggtggtg
      841 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      901 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
      961 ctgcccgtgc cctggcccac cctcgtgacc accctcagct acggcctgaa gtgcttcgcc
     1021 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1081 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1141 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcgg cttcaaggag
     1201 gacggcaaca tcctggggca caagctggag tacaacggcc ccagttgcag cgagcttgcc
     1261 gccaaacagc gcattcaggg tctcaccaac acccttcaag gctcaagcat caataccatc
     1321 cgccacatgg ttgccagtgg cttggcaatc agcgtcttgc ctgccactgc ccttaccgaa
     1381 aacgaccaca tgctgtttag cattatcccg tttgaaggca ccccgcctag ccgccgcgtt
     1441 gtcctggcct atcgccgcaa ctttgtccgc cctaaggccc tttcagccat gaaagccgca
     1501 attatgcaaa gccaactgca tggtgttagc ttcatctgcg attgactcga gcctctagaa
     1561 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     1621 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1681 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1741 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     1801 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     1861 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1921 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1981 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2041 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2701 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2941 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3001 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3061 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3121 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3181 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3241 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3301 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3361 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3421 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3481 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3541 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3601 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3661 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3721 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3781 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3841 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3901 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3961 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4021 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4081 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4141 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4201 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4261 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4321 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4381 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4441 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     4501 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt
     4561 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
     4621 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
     4681 caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact
     4741 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
     4801 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
     4861 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
     4921 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
     4981 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
     5041 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
     5101 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
     5161 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
     5221 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
     5281 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
     5341 agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
     5401 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
     5461 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
     5521 gtttagggaa c
//
