LOCUS       sequence-340570-        7411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid for a HyPer7 and tdTomato fusion under hSyn 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7411)
  AUTHORS   Li X, Zhang Y, Ai HW
  TITLE     Ratiometric Imaging of Mitochondrial Hydrogen Peroxide in 
            Abeta42-Mediated Neurotoxicity.
  JOURNAL   ACS Sens. 2022 Feb 28. doi: 10.1021/acssensors.1c01381.
  PUBMED    35226474
REFERENCE   2  (bases 1 to 7411)
  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"; journalName: "ACS Sens. 
            2022 Feb 28. doi: 10.1021/acssensors.1c01381."
COMMENT     Sequence Label: pAAV-hSyn-Mito-HyPerGRt
FEATURES             Location/Qualifiers
     source          1..7411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   76..205
                     /label=AAV2 ITR
     promoter        281..728
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             777..851
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             873..944
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     1577..1598
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1707..1728)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1968..1987)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2439..3866
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     misc_feature    3924..4512
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3977..3997)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4395..4406)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4515..4531)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4516..4532)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4561..4682
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4572..4591)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4626..4645
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4844..4973
                     /label=AAV2 ITR
     rep_origin      complement(5210..5798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5290..5309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5969..6760,6761..6829))
                     /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     6592..6611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6830..6934)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(6960..7411,1..4))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7097..7118)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7309..7328
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
       61 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
      121 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      181 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
      241 acgtagccat gctctaggaa gatccagggc cctgcgtatg agtgcaagtg ggttttagga
      301 ccaggatgag gcggggtggg ggtgcctacc tgacgaccgg ccccgaccca ctggacaagc
      361 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      421 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      481 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      541 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      601 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      661 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      721 gagcgcagtc caattcaagc tgctagcaag gatcccatcg attcgaattc gccaccatgt
      781 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc
      841 cgcgcgccaa gatccattcg ttgggggatc tgtccgtcct gacgccgctg ctgctgcggg
      901 gcttgacagg ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttggggg
      961 ataccggtca cctggctaat gaggagcaaa acgaattgga aggtgctttt aagctgggct
     1021 tgatctttac cgtcgccccg tacttgctgc ctaagttgat cgtcagcctg cgccgcaccg
     1081 ccccgaagat gcctttgatg ttggaggaga attacaccca cacccttacc gagtcactga
     1141 aacgcggcga cgttgacgcc atcattgtgg cagagccgtt tcaagagccg ggtattgtga
     1201 ctgaaccttt gtatgacgag ccgtttttcg ttatcgttcc gaagggccat agcttcgagg
     1261 agttggatgc cgttagccct cgcatgttgg gcgaagaaca agttctgctt ctgaccgagg
     1321 gcaactgtat gcgtgaccag gtgcttagct ccgccggctt caacagcaac aacgtctata
     1381 tcatggccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacgtcg
     1441 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1501 ccgtgctgct gcccgacaac cactacctga gcttccagtc cgtcctgagc aaagacccca
     1561 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1621 gcatggacga gctgtacaac gtggatggcg gtagcggtgg caccggcagc aagggcgagg
     1681 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1741 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagc
     1801 tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctcagct
     1861 acggcctgaa gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1921 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1981 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2041 agggcatcgg cttcaaggag gacggcaaca tcctggggca caagctggag tacaacggcc
     2101 ccagttgcag cgagcttgcc gccaaacagc gcattcaggg tctcaccaac acccttcaag
     2161 gctcaagcat caataccatc cgccacatgg ttgccagtgg cttggcaatc agcgtcttgc
     2221 ctgccactgc ccttaccgaa aacgaccaca tgctgtttag cattatcccg tttgaaggca
     2281 ccccgcctag ccgccgcgtt gtcctggcct atcgccgcaa ctttgtccgc cctaaggccc
     2341 tttcagccat gaaagccgca attatgcaaa gccaactgca tggtgttagc ttcatctgcg
     2401 atctcgaggg cggttctggt ggtggcggta gcggtggtgt gagcaagggc gaggaggtca
     2461 tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg
     2521 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     2581 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg
     2641 gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc
     2701 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga
     2761 cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca
     2821 acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg
     2881 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga
     2941 aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc
     3001 aactgcccgg ctactactac gtggacacca agctggacat cacctcccac aacgaggact
     3061 acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg
     3121 gcaccggcag caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg
     3181 ccgtcatcaa agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg
     3241 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     3301 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca
     3361 tgtacggctc caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt
     3421 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga
     3481 ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg
     3541 gcaccaactt cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     3601 ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga
     3661 agctgaagga cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc
     3721 ccgtgcaact gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg
     3781 aggactacac catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt
     3841 acggcatgga cgagctgtac aagtgagcta gccggcggcc tctagaacta tagtgagtcg
     3901 tattacgtag aaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3961 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4021 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4081 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4141 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4201 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4261 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4321 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4381 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4441 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4501 ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg gccgcttcga gcagacatga
     4561 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4621 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4681 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     4741 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat
     4801 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     4861 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4921 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt
     4981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     6961 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     7021 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     7081 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7141 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     7201 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     7261 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7321 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7381 acccgccgcg cttaatgcgc cgctacaggg c
//
