LOCUS       sequence-481762-        5798 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Mammalian neurons SypHer3s expression targeted to the mitochondrial 
            matrix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5798)
  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 5798)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 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_SypHer3s_mito
FEATURES             Location/Qualifiers
     source          1..5798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      359..947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     848..867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1009..1138
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        1153..1456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1464..1911
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1924..1998
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2020..2091
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     complement(2818..2839)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3079..3098)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3553..4029
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3752..3771)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4069..4209
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4069..4198
                     /label=AAV2 ITR
     rep_origin      4284..4739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4371..4390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4581..4602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4756..4775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4875..4897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4935..4953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5021..5125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5126..5194,5195..5798,1..188)
                     /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(5344..5363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
       61 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      121 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
      181 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
      241 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      301 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
      361 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
      421 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
      481 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
      541 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
      601 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
      661 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
      721 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
      781 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      841 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      901 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      961 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     1021 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     1081 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     1141 ggccgcacgc gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1201 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1261 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1321 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1381 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1441 catcgctatt accatggctc gagagtgcaa gtgggtttta ggaccaggat gaggcggggt
     1501 gggggtgcct acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc
     1561 caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc
     1621 gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac
     1681 cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt
     1741 cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg
     1801 cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg
     1861 cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca ggaattcgcc
     1921 accatgtccg tcctgacgcc gctgctgctg cggggcttga caggctcggc ccggcggctc
     1981 ccagtgccgc gcgccaagat ccattcgttg ggggatctgt ccgtcctgac gccgctgctg
     2041 ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc cgcgcgccaa gatccattcg
     2101 ttgggggatc catccggacc gctgcacatt ggtttgattc ccacagttgg accgtacctg
     2161 ctaccgcata ttatccctat gctgcaccag acctttccaa agctggaaat gtatctgcat
     2221 gaagcacaga cccaccagtt actggcgcaa ctggacagcg gcaaactcga ttgcgtgatc
     2281 ctcgcgctgg tgaaagagag cgaagcattc attgaagtgc cgttgtttga tgagccaatg
     2341 ttgctggcta tctatgaaga tcacccgtgg gcgaaccgcg aatgcgtacc gatggccgat
     2401 ctggcagggg aaaaactgct gatgctggaa gatggtcact ctttgcgcga tcaggcaatg
     2461 tccgccggct tcaacagcga caacgtctat atcatggccg acaagcagaa gaacggcatc
     2521 aaggccaact tcaagatccg ccacaacgtc gaggacggca gcgtgcagct cgccgaccac
     2581 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2641 agcttccagt ccgtcctgag caaagacccc aacgagaagc gcgatcacat ggtcctgcag
     2701 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa cgtggatggc
     2761 ggtagcggtg gcaccggcag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2821 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2881 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     2941 ccctggccca ccctcgtgac caccctcggc tacggcctga agtgcttcgc ccgctacccc
     3001 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3061 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3121 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg gcttcaagga ggacggcaac
     3181 atcctggggc acaagctgga gtacaacggc accggtttct gttttgaagc cggggcggat
     3241 gaagatacac acttccgcgc gaccagcctg gaaactctgc gcaacatggt ggcggcaggt
     3301 agcgggatca ctttactgcc agcgctggct gtgccgccgg agcgcaaacg cgatggggtt
     3361 gtttatctgc cgtgcattaa gccggaacca cgccgcacta ttggcctggt ttatcgtcct
     3421 ggctcaccgc tgcgcagccg ctatgagcag ctggcagagg ccatccgcgc aagaatggat
     3481 ggccatttcg ataaagtttt aaaacaggcg gtttaaaagc ttgcctcgag cagcgctgct
     3541 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3601 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3661 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3721 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3781 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     3841 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     3901 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     3961 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     4021 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     4081 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4141 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4201 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4261 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4441 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4621 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4681 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4741 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4801 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4861 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4921 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4981 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5041 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5101 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5161 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5221 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5281 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5341 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5401 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5461 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5521 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5581 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5641 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5701 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5761 taaagttgca ggaccacttc tgcgctcggc ccttccgg
//
