LOCUS       sequence-471083-        7611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of human Junctophilin3 N-terminally tagged with 
            HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7611)
  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 7611)
  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 HaloTag-Jph3 WPRE
FEATURES             Location/Qualifiers
     source          1..7611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(83..538)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(220..241)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   610..739
                     /label=AAV2 ITR
     intron          1332..1464
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1447..1466
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1488..1497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1494..2384
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    4683..5271
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4736..4756)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5154..5165)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5295..5416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5306..5325)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5360..5379
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5578..5707
                     /label=AAV2 ITR
     rep_origin      complement(5944..6532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6024..6043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6703..7494,7495..7563))
                     /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     7326..7345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7564..7611,1..57))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
       61 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
      121 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
      181 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
      241 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
      301 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
      361 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
      421 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
      481 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
      541 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      601 ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
      661 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
      721 tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag
      781 ccatgctcta ggaagatccg agctccagtg tgctggaatt cgcccttttc agtatttaaa
      841 ttgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      901 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      961 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
     1021 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
     1081 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
     1141 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
     1201 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagc tgtgctcctg ggcaccgcgc
     1261 agtccgcccc cgcggctcct ggccagacca cccctaggac cccctgcccc aagtcgcagc
     1321 cgttggatca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     1381 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     1441 tccactttgc ctttctctcc acaggtgatg atatcactag tagatctgcc accatggcag
     1501 aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc
     1561 actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga
     1621 cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg
     1681 ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg
     1741 accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg
     1801 tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg
     1861 tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag
     1921 aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca
     1981 tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg
     2041 aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt
     2101 ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg
     2161 aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc
     2221 caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca
     2281 aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg
     2341 gcagcgagat cgcgcgctgg ctgtcgacgc tcgagatttc cggcggatcc ggcggctccg
     2401 gaggcagcgg cggttctgga ggtgctagca tgtccagtgg gggcaggttt aattttgacg
     2461 acggagggtc ctactgtgga ggctgggagg acggcaaggc gcacggccat ggcgtctgca
     2521 ccggccccaa gggccaaggc gaatacaccg gctcgtggag ccacggcttc gaggtgctgg
     2581 gcgtctacac ctggcccagc ggcaacacgt accagggcac ctgggcgcag ggcaagcgcc
     2641 acggcatcgg cctggagagc aaggggaagt gggtgtacaa gggcgagtgg acgcacggat
     2701 tcaaggggcg ctacggggtg cgggagtgcg cgggcaacgg ggccaaatac gaagggacct
     2761 ggagcaacgg gctgcaggac ggctacggga ccgagaccta ctcggacgga gggacctacc
     2821 agggccagtg ggtcggtggc atgcgccagg gctacggcgt ccggcagagc gtcccgtatg
     2881 gcatggccgc ggtcatccgc tcacccctga ggacgtccat caactccctg cgcagcgagc
     2941 acaccaacgg cacggcgctg catcccgacg cctctccggc ggtggccggc agcccggccg
     3001 tgtcccgcgg gggcttcgtg ctcgtggccc acagtgactc cgagatcctc aagagcaaga
     3061 agaaggggct gtttcggcgc tcgctgctga gtgggctgaa gctgcgcaag tcggagtcca
     3121 agagcagcct ggccagccaa cgcagcaagc agagctcctt tcgcagcgag gcgggcatga
     3181 gcaccgtcag ctccacggcc agcgacatcc actccaccat cagcctgggc gaggctgagg
     3241 ccgagctggc ggtcatcgag gacgacatcg acgccaccac caccgagacc tacgtgggcg
     3301 agtggaagaa cgacaaacgc tccggcttcg gcgtgagcca gcgctcggac gggctcaagt
     3361 acgagggcga gtgggccagc aaccggcgcc atggctacgg ctgcatgacc ttcccggacg
     3421 gcaccaagga ggagggcaag tacaagcaga acatcctcgt cggcggcaag cgcaagaacc
     3481 tcatccccct gcgggccagc aagatccgcg agaaggtgga ccgcgccgtt gaggccgctg
     3541 agcgggccgc caccaccgcc aagcagaagg ctgagatcgc ggcttccagg acctcccact
     3601 ctcgggcaaa ggccgaggca gccctcacag cagctcagaa agcccaggag gaggcgcgga
     3661 tcgccaggat cactgccaaa gagttctccc cttccttcca gcaccgggaa aacgggctgg
     3721 agtaccagag gccgaagcgt cagacctcct gtgacgacat cgaggtgctg tccaccggga
     3781 cacccctgca gcaggagagc cccgagctgt accgcaaggg caccactccc tccgacctga
     3841 ccaccgacga cagccccctg cagagcttcc ccaccagccc cgcggccacc ccgccgcccg
     3901 cgcccgccgc caggaacaag gtcgcccact tctcgaggca ggtgtcggtg gacgaggagc
     3961 ggggcgggga catccagatg ctcctggagg gccgggccgg ggactgcgcc cgcagcagct
     4021 ggggcgagga gcaggccggg ggctccaggg gtgtccgcag cggtgccctg cgcggcggcc
     4081 tgctcgtgga tgacttccgc acccgaggtt cgggccgcaa gcagcccggg aaccccaagc
     4141 cgcgggagcg gcggacggag tcaccccccg tgttcacgtg gacttcccac caccgggcca
     4201 gcaaccacag ccccggaggc tccaggctgc tggagctgca ggaggagaag ctgagcaact
     4261 accggatgga gatgaaaccc ttgctgagga tggagacgca tccccagaaa agacgctaca
     4321 gcaagggcgg cgcctgccgg ggcttggggg acgaccaccg cctcgaggac cggggcttcg
     4381 gggtgcagag actgcggtcc aaggcccaga acaaggagaa cttcaggccg gcctcctccg
     4441 cggagcccgc cgtgcagaaa ctggcgagcc tgcggctggg cggggccgag ccccggttgc
     4501 tgcgttggga cttgaccttc tccccgcccc agaaatcctt gcctgtcgct ctagagtccg
     4561 acgaggagaa tggggatgag ctcaagtcca gtacgggctc agcgcctatc ctggtggtca
     4621 tggtgatctt gctcaacatc ggagtcgcca ttctgtttat taactttttc atctgaaagc
     4681 ttaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4741 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4801 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4861 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4921 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4981 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5041 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     5101 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5161 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5221 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cgcggccgct
     5281 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     5341 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     5401 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     5461 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     5521 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     5581 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     5641 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     5701 cgcgcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc t
//
