LOCUS       sequence-483492-        5834 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  Mammalian neurons HyPer7 expression targeted to the mitochondrial 
            matrix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  AUTHORS   Kelmanson IV, Shokhina AG, Kotova DA, Pochechuev MS, Ivanova AD, 
            Kostyuk AI, Panova AS, Borodinova AA, Solotenkov MA, Stepanov EA, 
            Raevskii RI, Moshchenko AA, Pak VV, Ermakova YG, van Belle GJC, 
            Tarabykin V, Balaban PM, Fedotov IV, Fedotov AB, Conrad M, Bogeski 
            I, Katschinski DM, Doeppner TR, Bahr M, Zheltikov AM, Belousov VV, 
            Bilan DS
  TITLE     In vivo dynamics of acidosis and oxidative stress in the acute phase
            of an ischemic stroke in a rodent model.
  JOURNAL   Redox Biol. 2021 Nov 3;48:102178. doi: 10.1016/j.redox.2021.102178.
  PUBMED    34773835
REFERENCE   2  (bases 1 to 5834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Redox 
            Biol."; date: "2021-11-3"; volume: "48:102178. doi"; pages: " 
            10.1016/j.redox.2021.102178"
COMMENT     Sequence Label: pAAV_hSyn1_HyPer7_mito
FEATURES             Location/Qualifiers
     source          1..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        145..448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        456..903
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             916..990
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1012..1083
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     1716..1737
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1846..1867)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2107..2126)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2581..3057
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2780..2799)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3097..3237
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3097..3226
                     /label=AAV2 ITR
     rep_origin      3312..3767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3399..3418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3609..3630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3784..3803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3903..3925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3963..3981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4049..4153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4154..4222,4223..5014)
                     /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     complement(4372..4391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5185..5773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5674..5693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgttac ataacttacg gtaaatggcc cgcctggctg
      181 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      301 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      361 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      421 ctacgtatta gtcatcgcta ttaccatggc tcgagagtgc aagtgggttt taggaccagg
      481 atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca
      541 acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg
      601 cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc
      661 ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg
      721 caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg
      781 caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc
      841 gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg
      901 caggaattcg ccaccatgtc cgtcctgacg ccgctgctgc tgcggggctt gacaggctcg
      961 gcccggcggc tcccagtgcc gcgcgccaag atccattcgt tgggggatct gtccgtcctg
     1021 acgccgctgc tgctgcgggg cttgacaggc tcggcccggc ggctcccagt gccgcgcgcc
     1081 aagatccatt cgttggggga taccggtcac ctggctaatg aggagcaaaa cgaattggaa
     1141 ggtgctttta agctgggctt gatctttacc gtcgccccgt acttgctgcc taagttgatc
     1201 gtcagcctgc gccgcaccgc cccgaagatg cctttgatgt tggaggagaa ttacacccac
     1261 acccttaccg agtcactgaa acgcggcgac gttgacgcca tcattgtggc agagccgttt
     1321 caagagccgg gtattgtgac tgaacctttg tatgacgagc cgtttttcgt tatcgttccg
     1381 aagggccata gcttcgagga gttggatgcc gttagccctc gcatgttggg cgaagaacaa
     1441 gttctgcttc tgaccgaggg caactgtatg cgtgaccagg tgcttagctc cgccggcttc
     1501 aacagcaaca acgtctatat catggccgac aagcagaaga acggcatcaa ggccaacttc
     1561 aagatccgcc acaacgtcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1621 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cttccagtcc
     1681 gtcctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1741 gccgccggga tcactctcgg catggacgag ctgtacaacg tggatggcgg tagcggtggc
     1801 accggcagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1861 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1921 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1981 ctcgtgacca ccctcagcta cggcctgaag tgcttcgccc gctaccccga ccacatgaag
     2041 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2101 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2161 gtgaaccgca tcgagctgaa gggcatcggc ttcaaggagg acggcaacat cctggggcac
     2221 aagctggagt acaacggccc cagttgcagc gagcttgccg ccaaacagcg cattcagggt
     2281 ctcaccaaca cccttcaagg ctcaagcatc aataccatcc gccacatggt tgccagtggc
     2341 ttggcaatca gcgtcttgcc tgccactgcc cttaccgaaa acgaccacat gctgtttagc
     2401 attatcccgt ttgaaggcac cccgcctagc cgccgcgttg tcctggccta tcgccgcaac
     2461 tttgtccgcc ctaaggccct ttcagccatg aaagccgcaa ttatgcaaag ccaactgcat
     2521 ggtgttagct tcatctgcga ttgaaagctt gcctcgagca gcgctgctcg agagatctac
     2581 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     2641 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     2701 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     2761 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     2821 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     2881 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     2941 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     3001 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     3061 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3121 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3181 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3241 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3301 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3361 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3421 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3481 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3541 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3601 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3661 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3721 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3781 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3841 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3901 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3961 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4021 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4081 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4141 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4201 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4261 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4321 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4381 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4441 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4501 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4561 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4621 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4681 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4741 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4801 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4861 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4921 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4981 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5041 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5101 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5161 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5221 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5281 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5341 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5401 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5461 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5521 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5581 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5641 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5701 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5761 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5821 ttttgctcac atgt
//
