LOCUS       sequence-471241-        7389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of human Junctophilin3 N-terminally tagged with 
            mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7389)
  AUTHORS   Benedetti L, Fan R, Weigel AV, Moore AS, Houlihan PR, Kittisopikul 
            M, Park G, Petruncio A, Hubbard PM, Pang S, Xu CS, Hess HF, Saalfeld
            S, Rangaraju V, Clapham DE, De Camilli P, Ryan TA, 
            Lippincott-Schwartz J
  TITLE     Periodic ER-plasma membrane junctions support long-range Ca(2+) 
            signal integration in dendrites.
  JOURNAL   Cell. 2025 Jan 23;188(2):484-500.e22. doi: 
            10.1016/j.cell.2024.11.029. Epub 2024 Dec 20.
  PUBMED    39708809
REFERENCE   2  (bases 1 to 7389)
  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.cell.2024.11.029"; journalName: "Cell"; date: 
            "2025-01-23- 23"; volume: "188"; issue: "2"; pages: "484-500.e22"
COMMENT     Sequence Label: pAAV hSyn INTRON mCherry-Jph3 WPRE
FEATURES             Location/Qualifiers
     source          1..7389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(996..1787,1788..1856))
                     /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     1619..1638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1857..1961)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1987..2442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2124..2145)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   2514..2643
                     /label=AAV2 ITR
     promoter        2725..3172
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          3194..3326
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3309..3328
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3340..3349
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3346..4053
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3491..3509)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3759..3778
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6340..6928
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6393..6413)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6811..6822)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6931..6947)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6932..6948)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6977..7098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6988..7007)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7042..7061
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   7260..7389
                     /label=AAV2 ITR
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1981 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2041 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2101 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2161 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2221 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2281 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2341 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2401 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     2461 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
     2521 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     2581 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2641 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     2701 tctctgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga tgaggcgggg
     2761 tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc
     2821 ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg
     2881 cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca
     2941 ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct
     3001 tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag
     3061 gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct
     3121 gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agtcgaattc
     3181 aagctgctag caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     3241 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     3301 catccacttt gcctttctct ccacaggtga taaggatccg ccaccatggt gagcaagggc
     3361 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     3421 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     3481 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     3541 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     3601 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     3661 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     3721 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     3781 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3841 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     3901 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     3961 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     4021 cactccaccg gcggcatgga cgagctgtac aagagatctc gagctcaagc ttcgaattct
     4081 atgtccagtg ggggcaggtt taattttgac gacggagggt cctactgtgg aggctgggag
     4141 gacggcaagg cgcacggcca tggcgtctgc accggcccca agggccaagg cgaatacacc
     4201 ggctcgtgga gccacggctt cgaggtgctg ggcgtctaca cctggcccag cggcaacacg
     4261 taccagggca cctgggcgca gggcaagcgc cacggcatcg gcctggagag caaggggaag
     4321 tgggtgtaca agggcgagtg gacgcacgga ttcaaggggc gctacggggt gcgggagtgc
     4381 gcgggcaacg gggccaaata cgaagggacc tggagcaacg ggctgcagga cggctacggg
     4441 accgagacct actcggacgg agggacctac cagggccagt gggtcggtgg catgcgccag
     4501 ggctacggcg tccggcagag cgtcccgtat ggcatggccg cggtcatccg ctcacccctg
     4561 aggacgtcca tcaactccct gcgcagcgag cacaccaacg gcacggcgct gcatcccgac
     4621 gcctctccgg cggtggccgg cagcccggcc gtgtcccgcg ggggcttcgt gctcgtggcc
     4681 cacagtgact ccgagatcct caagagcaag aagaaggggc tgtttcggcg ctcgctgctg
     4741 agtgggctga agctgcgcaa gtcggagtcc aagagcagcc tggccagcca acgcagcaag
     4801 cagagctcct ttcgcagcga ggcgggcatg agcaccgtca gctccacggc cagcgacatc
     4861 cactccacca tcagcctggg cgaggctgag gccgagctgg cggtcatcga ggacgacatc
     4921 gacgccacca ccaccgagac ctacgtgggc gagtggaaga acgacaaacg ctccggcttc
     4981 ggcgtgagcc agcgctcgga cgggctcaag tacgagggcg agtgggccag caaccggcgc
     5041 catggctacg gctgcatgac cttcccggac ggcaccaagg aggagggcaa gtacaagcag
     5101 aacatcctcg tcggcggcaa gcgcaagaac ctcatccccc tgcgggccag caagatccgc
     5161 gagaaggtgg accgcgccgt tgaggccgct gagcgggccg ccaccaccgc caagcagaag
     5221 gctgagatcg cggcttccag gacctcccac tctcgggcaa aggccgaggc agccctcaca
     5281 gcagctcaga aagcccagga ggaggcgcgg atcgccagga tcactgccaa agagttctcc
     5341 ccttccttcc agcaccggga aaacgggctg gagtaccaga ggccgaagcg tcagacctcc
     5401 tgtgacgaca tcgaggtgct gtccaccggg acacccctgc agcaggagag ccccgagctg
     5461 taccgcaagg gcaccactcc ctccgacctg accaccgacg acagccccct gcagagcttc
     5521 cccaccagcc ccgcggccac cccgccgccc gcgcccgccg ccaggaacaa ggtcgcccac
     5581 ttctcgaggc aggtgtcggt ggacgaggag cggggcgggg acatccagat gctcctggag
     5641 ggccgggccg gggactgcgc ccgcagcagc tggggcgagg agcaggccgg gggctccagg
     5701 ggtgtccgca gcggtgccct gcgcggcggc ctgctcgtgg atgacttccg cacccgaggt
     5761 tcgggccgca agcagcccgg gaaccccaag ccgcgggagc ggcggacgga gtcacccccc
     5821 gtgttcacgt ggacttccca ccaccgggcc agcaaccaca gccccggagg ctccaggctg
     5881 ctggagctgc aggaggagaa gctgagcaac taccggatgg agatgaaacc cttgctgagg
     5941 atggagacgc atccccagaa aagacgctac agcaagggcg gcgcctgccg gggcttgggg
     6001 gacgaccacc gcctcgagga ccggggcttc ggggtgcaga gactgcggtc caaggcccag
     6061 aacaaggaga acttcaggcc ggcctcctcc gcggagcccg ccgtgcagaa actggcgagc
     6121 ctgcggctgg gcggggccga gccccggttg ctgcgttggg acttgacctt ctccccgccc
     6181 cagaaatcct tgcctgtcgc tctagagtcc gacgaggaga atggggatga gctcaagtcc
     6241 agtacgggct cagcgcctat cctggtggtc atggtgatct tgctcaacat cggagtcgcc
     6301 attctgttta ttaacttttt catctgaaag cttatcgata atcaacctct ggattacaaa
     6361 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     6421 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     6481 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     6541 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     6601 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     6661 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     6721 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     6781 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     6841 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     6901 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg
     6961 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     7021 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     7081 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     7141 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     7201 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     7261 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     7321 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     7381 agcgcgcag
//
