LOCUS       sequence-323579-        4863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ultrasensitive hydrogen peroxide indicator 
            HyPer7 for optical imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4863)
  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 4863)
  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: 
            "10.1016/j.cmet.2020.02"; journalName: "Cell Metab"; date: 
            "2020-03-3- 3"; volume: "31"; issue: "3"; pages: "642-653.e6"
COMMENT     Sequence Label: pQE30-HyPer7
FEATURES             Location/Qualifiers
     source          1..4863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     RBS             65..76
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             95..112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     745..766
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(875..896)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1136..1155)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     terminator      1610..1704
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1748..2407
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1814..1833)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2472..2558
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2605..2627)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2713..2853
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2768..2787
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2868..2885)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3039..3627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3119..3138)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3798..4589,4590..4658))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4421..4440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4659..4763)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4831..4849
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaatttc acacagaatt
       61 cattaaagag gagaaattaa ctatgagagg atcgcatcac catcaccatc acggatcccg
      121 ggaattcgcc accatgcacc tggctaatga ggagcaaaac gaattggaag gtgcttttaa
      181 gctgggcttg atctttaccg tcgccccgta cttgctgcct aagttgatcg tcagcctgcg
      241 ccgcaccgcc ccgaagatgc ctttgatgtt ggaggagaat tacacccaca cccttaccga
      301 gtcactgaaa cgcggcgacg ttgacgccat cattgtggca gagccgtttc aagagccggg
      361 tattgtgact gaacctttgt atgacgagcc gtttttcgtt atcgttccga agggccatag
      421 cttcgaggag ttggatgccg ttagccctcg catgttgggc gaagaacaag ttctgcttct
      481 gaccgagggc aactgtatgc gtgaccaggt gcttagctcc gccggcttca acagcaacaa
      541 cgtctatatc atggccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
      601 caacgtcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      661 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc ttccagtccg tcctgagcaa
      721 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      781 cactctcggc atggacgagc tgtacaacgt ggatggcggt agcggtggca ccggcagcaa
      841 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      901 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      961 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1021 cctcagctac ggcctgaagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1081 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1141 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1201 cgagctgaag ggcatcggct tcaaggagga cggcaacatc ctggggcaca agctggagta
     1261 caacggcccc agttgcagcg agcttgccgc caaacagcgc attcagggtc tcaccaacac
     1321 ccttcaaggc tcaagcatca ataccatccg ccacatggtt gccagtggct tggcaatcag
     1381 cgtcttgcct gccactgccc ttaccgaaaa cgaccacatg ctgtttagca ttatcccgtt
     1441 tgaaggcacc ccgcctagcc gccgcgttgt cctggcctat cgccgcaact ttgtccgccc
     1501 taaggccctt tcagccatga aagccgcaat tatgcaaagc caactgcatg gtgttagctt
     1561 catctgcgat tgactcgagg ccgactgtaa gcttaattag ctgagcttgg actcctgttg
     1621 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     1681 cgggcgtttt ttattggtga gaatccaagc tagcttggcg agattttcag gagctaagga
     1741 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1801 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1861 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1921 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa
     1981 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     2041 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     2101 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     2161 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     2221 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     2281 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     2341 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     2401 ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta
     2461 gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2521 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccctctaga gctgcctcgc
     2581 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2641 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2701 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     2761 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2821 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     2881 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2941 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3001 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3061 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3121 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3181 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3241 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3301 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3361 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3421 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3481 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3541 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3601 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3661 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3721 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3781 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3841 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3901 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3961 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4021 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4081 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4141 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4201 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4261 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4321 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4381 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4441 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4501 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4561 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4621 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4681 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4741 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     4801 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
     4861 cac
//
