LOCUS       sequence-265174-        5567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cytoplasm 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 5567)
  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 5567)
  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: pCS2+HyPer7-NES
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     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             1549..1575
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    1616..1750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1640..1659)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1694..1713
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1863..1883)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1863..1881)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1902..1918)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1902..1918)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1915..1937)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1926..1942
                     /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)."
     promoter        complement(join(1950..1956,1957..1974,1975..1980))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1995..2016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2133..2150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2304..2892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2384..2403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3063..3854,3855..3923))
                     /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     3686..3705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3924..4028)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4054..4509)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4191..4212)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4403..4422
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4583..5567
                     /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 ataccggtct gccccccctg
     1561 gagcggctga ccctgtaact cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     1621 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1681 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1741 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1801 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     1861 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     1921 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1981 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2041 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2101 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2461 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2701 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     2761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3901 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3961 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4021 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     4081 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4141 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4201 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4261 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4321 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     4381 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     4441 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4501 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     4561 cctcttcgct attacgccag tcgaccatag ccaattcaat atggcgtata tggactcatg
     4621 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg
     4681 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg
     4741 ccaattcaat atggcggact tggcactgtg ccaactgggg aggggtctac ttggcacggt
     4801 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt
     4861 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata
     4921 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat
     4981 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc
     5041 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat
     5101 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg
     5161 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg
     5221 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt
     5281 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc
     5341 tattaatagt aacttggcaa gtacattact attggaagga cgccagggta cattggcagt
     5401 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt
     5461 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta
     5521 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
