LOCUS       sequence-483533-        6989 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  Mammalian astrocytes HyPer7 expression targeted to the mitochondrial
            matrix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6989)
  AUTHORS   Kotova DA, Ivanova AD, Pochechuev MS, Kelmanson IV, Khramova YV, 
            Tiaglik A, Sudoplatov MA, Trifonova AP, Fedotova A, Morozova K, 
            Katrukha VA, Sergeeva AD, Raevskii RI, Pestriakova MP, Solotenkov 
            MA, Stepanov EA, Tsopina AS, Moshchenko AA, Shestopalova M, Zalygin 
            A, Fedotov IV, Fedotov AB, Oleinikov V, Belousov VV, Semyanov A, 
            Brazhe N, Zheltikov AM, Bilan DS
  TITLE     Hyperglycemia exacerbates ischemic stroke not through increased 
            generation of hydrogen peroxide.
  JOURNAL   Free Radic Biol Med. 2023 Nov 1;208:153-164. doi: 
            10.1016/j.freeradbiomed.2023.08.004. Epub 2023 Aug 4.
  PUBMED    37543166
REFERENCE   2  (bases 1 to 6989)
  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: "Free Radic 
            Biol Med."; date: "2023-11-1"; volume: "208:153-164. doi"; pages: " 
            10.1016/j.freeradbiomed.2023.08.004. Epub 2023 Aug 4"
COMMENT     Sequence Label: pAAV_GFAP_HyPer7_mito
FEATURES             Location/Qualifiers
     source          1..6989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             2071..2145
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2167..2238
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     2871..2892
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3001..3022)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3262..3281)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3736..4212
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3935..3954)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4252..4392
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4252..4381
                     /label=AAV2 ITR
     rep_origin      4467..4922
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4554..4573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4764..4785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4939..4958)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5058..5080
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5118..5136)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5204..5308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5309..5377,5378..6169)
                     /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"
     rep_origin      6340..6928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6829..6848
                     /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 gcgtcttaag gctgggaggt tgaggctgca gtgagtcgtg
      181 gttgcgccac tgcactccag cctgggcaac agtgagaccc tgtctcaaaa gacaaaaaaa
      241 aaaaaaaaaa aagaacatat cctggtgtgg agtaggggac gctgctctga cagaggctcg
      301 ggggcctgag ctggctctgt gagctgggga ggaggcagac agccaggcct tgtctgcaag
      361 cagacctggc agcattgggc tggccgcccc ccagggcctc ctcttcatgc ccagtgaatg
      421 actcaccttg gcacagacac aatgttcggg gtgggcacag tgcctgcttc ccgccgcacc
      481 ccagcccccc tcaaatgcct tccgagaagc ccattgagca gggggcttgc attgcacccc
      541 agcctgacag cctggcatct tgggataaaa gcagcacagc cccctagggg ctgcccttgc
      601 tgtgtggcgc caccggcggt ggagaacaag gctctattca gcctgtgccc aggaaagggg
      661 atcaggggat gcccaggcat ggacagtggg tggcaggggg ggagaggagg gctgtctgct
      721 tcccagaagt ccaaggacac aaatgggtga ggggactggg cagggttctg accctgtggg
      781 accagagtgg agggcgtaga tggacctgaa gtctccaggg acaacagggc ccaggtctca
      841 ggctcctagt tgggcccagt ggctccagcg tttccaaacc catccatccc cagaggttct
      901 tcccatctct ccaggctgat gtgtgggaac tcgaggaaat aaatctccag tgggagacgg
      961 aggggtggcc agggaaacgg ggcgctgcag gaataaagac gagccagcac agccagctca
     1021 tgtgtaacgg ctttgtggag ctgtcaaggc ctggtctctg ggagagaggc acagggaggc
     1081 cagacaagga aggggtgacc tggagggaca gatccagggg ctaaagtcct gataaggcaa
     1141 gagagtgccg gccccctctt gccctatcag gacctccact gccacataga ggccatgatt
     1201 gacccttaga caaagggctg gtgtccaatc ccagccccca gccccagaac tccagggaat
     1261 gaatgggcag agagcaggaa tgtgggacat ctgtgttcaa gggaaggact ccaggagtct
     1321 gctggaaatg aggcctagta ggaaatgagg tggcccttga gggtacagaa caggttcatt
     1381 cttcgccaaa ttcccagcac cttgcaggca cttacagctg agtgagataa tgcctgggtt
     1441 atgaaatcaa aaagttggaa agcaggtcag aggtcatctg gtacagccct tccttccctt
     1501 tttttttttt ttttttgtga gacaaggtct ctctctgttg cccaggctgg agtggcgcaa
     1561 acacagctca ctgcagcctc aacctactgg gctcaagcaa tcctccagcc tcagcctccc
     1621 aaagtgctgg gattacaagc atgagccacc ccactcagcc ctttccttcc tttttaattg
     1681 atgcataata attgtaagta ttcatcatgg tccaaccaac cctttcttga cccaccttcc
     1741 tagagagagg gtcctcttgc ttcagcggtc agggccccag acccatggtc tggctccagg
     1801 taccacctgc ctcatgcagg agttggcgtg cccaggaagc tctgcctctg ggcacagtga
     1861 cctcagtggg gtgaggggag ctctccccat agctgggctg cggcccaacc ccaccccctc
     1921 aggctatgcc agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt
     1981 cataaagccc tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat
     2041 cacctccgct gctcgcggga attcgccacc atgtccgtcc tgacgccgct gctgctgcgg
     2101 ggcttgacag gctcggcccg gcggctccca gtgccgcgcg ccaagatcca ttcgttgggg
     2161 gatctgtccg tcctgacgcc gctgctgctg cggggcttga caggctcggc ccggcggctc
     2221 ccagtgccgc gcgccaagat ccattcgttg ggggataccg gtcacctggc taatgaggag
     2281 caaaacgaat tggaaggtgc ttttaagctg ggcttgatct ttaccgtcgc cccgtacttg
     2341 ctgcctaagt tgatcgtcag cctgcgccgc accgccccga agatgccttt gatgttggag
     2401 gagaattaca cccacaccct taccgagtca ctgaaacgcg gcgacgttga cgccatcatt
     2461 gtggcagagc cgtttcaaga gccgggtatt gtgactgaac ctttgtatga cgagccgttt
     2521 ttcgttatcg ttccgaaggg ccatagcttc gaggagttgg atgccgttag ccctcgcatg
     2581 ttgggcgaag aacaagttct gcttctgacc gagggcaact gtatgcgtga ccaggtgctt
     2641 agctccgccg gcttcaacag caacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2701 atcaaggcca acttcaagat ccgccacaac gtcgaggacg gcagcgtgca gctcgccgac
     2761 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2821 ctgagcttcc agtccgtcct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2881 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caacgtggat
     2941 ggcggtagcg gtggcaccgg cagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3001 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3061 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     3121 gtgccctggc ccaccctcgt gaccaccctc agctacggcc tgaagtgctt cgcccgctac
     3181 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3241 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3301 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcggcttcaa ggaggacggc
     3361 aacatcctgg ggcacaagct ggagtacaac ggccccagtt gcagcgagct tgccgccaaa
     3421 cagcgcattc agggtctcac caacaccctt caaggctcaa gcatcaatac catccgccac
     3481 atggttgcca gtggcttggc aatcagcgtc ttgcctgcca ctgcccttac cgaaaacgac
     3541 cacatgctgt ttagcattat cccgtttgaa ggcaccccgc ctagccgccg cgttgtcctg
     3601 gcctatcgcc gcaactttgt ccgccctaag gccctttcag ccatgaaagc cgcaattatg
     3661 caaagccaac tgcatggtgt tagcttcatc tgcgattgaa agcttgcctc gagcagcgct
     3721 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     3781 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     3841 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     3901 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     3961 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     4021 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     4081 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     4141 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     4201 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     4261 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4321 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     4381 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4441 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     4501 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     4561 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4621 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4681 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4741 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     4801 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     4861 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     4921 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4981 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     5041 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     5101 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5161 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5221 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5281 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5341 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5401 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5461 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5521 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5581 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5641 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5701 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5761 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5821 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5881 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5941 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     6001 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     6061 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6121 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6181 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6241 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6301 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6361 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6421 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6481 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6541 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6601 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6661 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6721 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6781 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6841 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6901 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6961 ggccttttgc tggccttttg ctcacatgt
//
