LOCUS       sequence-471243-        7377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of mouse STIM1 internally tagged with HaloTag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7377)
  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 7377)
  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-STIM1 WPRE
FEATURES             Location/Qualifiers
     source          1..7377
                     /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"
     primer_bind     2336..2355
                     /label=F1ori-R
                     /note="F1 origin, reverse 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             3421..4311
                     /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    6328..6916
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6381..6401)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6799..6810)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6919..6935)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6920..6936)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6965..7086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6976..6995)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7030..7049
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   7248..7377
                     /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 gcyattamks 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 ccaccatgga tgtgtgcgcc
     3361 cgtcttgccc tgtggcttct ttgggggctc cttctgcatc agggccagag tggcggctcc
     3421 atggcagaaa tcggtactgg ctttccattc gacccccatt atgtggaagt cctgggcgag
     3481 cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt
     3541 aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc gacccatcgc
     3601 tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct gggttatttc
     3661 ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct ggaagaggtc
     3721 gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa gcgcaatcca
     3781 gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac ctgggacgaa
     3841 tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt cggccgcaag
     3901 ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg
     3961 ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag
     4021 ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg
     4081 ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg
     4141 ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct
     4201 aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac
     4261 ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg agatttccgg cggcggctcc
     4321 ctcagccata gtcacagtga aaagaataca ggagcgagct ccggggcgac ttctgaagag
     4381 tctaccgaag cagagttttg ccgaattgac aagcccctgt gccacagtga ggatgagaag
     4441 ctcagctttg aggccgtccg aaacatccat aagctgatgg atgacgatgc caatggtgat
     4501 gtggatgtgg aagaaagtga tgagttccta agggaagacc tcaattacca tgacccaaca
     4561 gtgaaacata gcaccttcca tggtgaggat aaacttatca gcgtggagga cctgtggaag
     4621 gcgtggaagt catcagaagt gtacaactgg actgtggatg aggtgataca gtggctcatt
     4681 acgtatgtgg agctgccaca gtatgaggaa accttccgga agttgcagct tactggccac
     4741 gccatgccaa ggcttgcagt aaccaacacc accatgacag ggactgtact gaagatgaca
     4801 gatcggagcc acaggcagaa gctgcagctg aaggccctgg acacagtgct gtttgggcct
     4861 cctctcttga ctcggcataa tcacctgaag gacttcatgc tggtggtgtc tatcgttatt
     4921 ggtgtgggtg gctgctggtt tgcctatatc cagaaccgtt actctaagga gcacatgaag
     4981 aaaatgatga aggatctgga agggttacac cgggctgagc agagtctgca tgaccttcag
     5041 gaaaggctgc acaaggccca ggaggagcac cgaactgtgg aagtagagaa ggtccacctg
     5101 gagaagaagc tgcgagatga gatcaacctt gccaagcagg aagctcagcg gctgaaggag
     5161 ctgagggagg gtactgagaa tgagaggagc cgtcaaaaat atgctgagga agagctggag
     5221 caggttcggg aggccttgag gaaagcagag aaggagctgg aatcacacag ctcatggtat
     5281 gctcctgagg ccctgcagaa gtggctgcag ctgacccatg aggtggaggt gcagtactac
     5341 aacatcaaga agcaaaatgc agagaggcag ctgctggtgg ccaaggaggg ggctgagaaa
     5401 ataaaaaaga agagaaacac gctttttggt accttccatg tggcccacag ctcttccctg
     5461 gatgatgtgg atcataaaat cctaactgct aagcaagctc tgagtgaggt gacagcggca
     5521 ctgagggagc gcctgcaccg atggcagcag atcgagatcc tctgcggttt ccagattgtc
     5581 aataaccccg gcatccactc cttggtggct gctctcaaca tcgaccccag ctggatgggc
     5641 agcacccgcc ctaaccccgc ccacttcatc atgactgacg atgtggatga catggatgag
     5701 gagattgtgt cgcccttgtc catgcagtcc cccagcctgc agagcagtgt ccggcagcgc
     5761 ctgacggagc cacagcttgg cctgggatct cagagggatt tgacccattc cgattcggag
     5821 tcctccctcc acatgagtga ccgccagcgt gtggccccca agcctcctca gatgggccgt
     5881 gctgcagatg aagctctcaa tgccatgcct tccaatggca gccatcggct gattgagggg
     5941 gtccatccag gatctctggt ggagaaactg cctgacagcc ctgctctggc caagaagaca
     6001 tttatggcgt tgaaccatgg cctagacaag gcccacagcc tgatggagct gaacccctca
     6061 gtcccacctg gtggctcccc acttttggat tcttcccatt ctcttagccc cagttcccca
     6121 gacccagaca cgccatctcc agttggggac aaccgagctc tgcagggtag ccgaaacaca
     6181 cgaatacccc acttggctgg caagaaggca atggctgagg aggataatgg ttccattggt
     6241 gaggagacag actccagtcc aggcaggaag aagtttcctc tcaaaatttt taagaagcct
     6301 cttaagaagt agtaaaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     6361 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6421 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6481 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6541 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6601 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6661 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6721 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6781 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6841 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6901 tgggccgcct ccccgcatcg ataccgtcga cctcgacccg ggcggccgct tcgagcagac
     6961 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     7021 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     7081 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     7141 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga
     7201 gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt
     7261 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     7321 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcag
//
