LOCUS       sequence-271548-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mitochondrial intermembrane space 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 5765)
  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 5765)
  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+IMS-HyPer7
FEATURES             Location/Qualifiers
     source          1..5765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        475..493
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     475..492
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      540..549
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      544..553
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1391..1412
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1521..1542)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1782..1801)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2257..2391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2281..2300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2335..2354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2504..2524)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2504..2522)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2543..2559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2543..2559)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2556..2578)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2567..2583
                     /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(2591..2597,2598..2615,2616..2621))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2636..2657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2774..2791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2945..3533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3025..3044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3704..4495,4496..4564))
                     /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     4327..4346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4565..4669)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4695..5150)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4832..4853)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5044..5063
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5221..5765,1..440)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
       61 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
      121 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
      181 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
      241 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
      301 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
      361 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
      421 gcaatgctcg tttagggaac cgccattctg cctggggacg tcggagcaag cttgatttag
      481 gtgacactat agaatacaag ctacttgttc tttttgcagg atcccatcga ttcgaattcg
      541 ccaccatggc ggctctgaag agttggctgt cgcgcagcgt aacttcattc ttcaggtaca
      601 gacagtgttt gtgtgttcct gttgtggcta actttaagaa gcggtgtttc tcagaattga
      661 taagaccatg gcacaaaact gtgacgattg gctttggagt aaccctgtgt gcggttccta
      721 ttaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccc gtcgccaccg
      781 gtcacctggc taatgaggag caaaacgaat tggaaggtgc ttttaagctg ggcttgatct
      841 ttaccgtcgc cccgtacttg ctgcctaagt tgatcgtcag cctgcgccgc accgccccga
      901 agatgccttt gatgttggag gagaattaca cccacaccct taccgagtca ctgaaacgcg
      961 gcgacgttga cgccatcatt gtggcagagc cgtttcaaga gccgggtatt gtgactgaac
     1021 ctttgtatga cgagccgttt ttcgttatcg ttccgaaggg ccatagcttc gaggagttgg
     1081 atgccgttag ccctcgcatg ttgggcgaag aacaagttct gcttctgacc gagggcaact
     1141 gtatgcgtga ccaggtgctt agctccgccg gcttcaacag caacaacgtc tatatcatgg
     1201 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac gtcgaggacg
     1261 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1321 tgctgcccga caaccactac ctgagcttcc agtccgtcct gagcaaagac cccaacgaga
     1381 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1441 acgagctgta caacgtggat ggcggtagcg gtggcaccgg cagcaagggc gaggagctgt
     1501 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1561 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     1621 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctc agctacggcc
     1681 tgaagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1741 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1801 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1861 tcggcttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ggccccagtt
     1921 gcagcgagct tgccgccaaa cagcgcattc agggtctcac caacaccctt caaggctcaa
     1981 gcatcaatac catccgccac atggttgcca gtggcttggc aatcagcgtc ttgcctgcca
     2041 ctgcccttac cgaaaacgac cacatgctgt ttagcattat cccgtttgaa ggcaccccgc
     2101 ctagccgccg cgttgtcctg gcctatcgcc gcaactttgt ccgccctaag gccctttcag
     2161 ccatgaaagc cgcaattatg caaagccaac tgcatggtgt tagcttcatc tgcgattgac
     2221 tcgagcctct agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt
     2281 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2341 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2401 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg
     2461 gccgcggcgc caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     2521 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2581 caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag
     2641 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2701 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2761 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2821 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2881 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2941 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3001 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3061 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3121 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3181 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3241 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3301 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3361 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3421 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3481 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3541 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3601 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3661 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3721 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3781 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3841 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3901 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3961 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4021 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4081 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4141 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4201 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4261 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4321 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4381 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4441 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4501 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4561 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4621 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4681 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4741 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4801 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4861 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4921 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4981 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5041 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5101 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     5161 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5221 gtcgaccgcc aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg
     5281 gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg
     5341 gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg
     5401 gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt
     5461 ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg
     5521 gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta
     5581 ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat atatgggttt
     5641 tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc
     5701 ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc
     5761 tattg
//
