LOCUS       sequence-308715-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of nucleus 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 5615)
  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 5615)
  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-NLS
FEATURES             Location/Qualifiers
     source          1..5615
                     /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             1552..1572
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1576..1596
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1600..1620
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1667..1801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1691..1710)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1745..1764
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1914..1934)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1914..1932)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1953..1969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1953..1969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1966..1988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1977..1993
                     /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(2001..2007,2008..2025,2026..2031))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2046..2067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2184..2201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2355..2943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2435..2454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3114..3905,3906..3974))
                     /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     3737..3756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3975..4079)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4105..4560)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4242..4263)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4454..4473
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4631..5615
                     /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 ataccggtga tccaaaaaag
     1561 aagagaaagg tagatccaaa aaagaagaga aaggtagatc caaaaaagaa gagaaaggta
     1621 taagcttgcc tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat
     1681 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1741 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1801 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1861 ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag
     1921 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1981 tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt
     2041 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2101 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2161 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2761 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2941 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3001 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3061 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3121 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3181 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3241 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3301 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3361 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3421 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3481 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3541 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3601 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3661 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3721 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3781 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3841 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3901 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3961 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4081 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     4141 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4201 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4261 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4321 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4381 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4441 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4501 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4561 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4621 tattacgcca gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
     4681 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
     4741 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
     4801 ggcggacctg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
     4861 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
     4921 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
     4981 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
     5041 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
     5101 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
     5161 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
     5221 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
     5281 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
     5341 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
     5401 cttggcaagt acattactat tggaagtacg ccagggtaca ttggcagtac tcccattgac
     5461 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
     5521 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     5581 gggaggtcta tataagcaat gctcgtttag ggaac
//
