LOCUS       sequence-471084-        7641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of human STIM2 internally tagged with HaloTag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7641)
  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 7641)
  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-STIM2 WPRE
FEATURES             Location/Qualifiers
     source          1..7641
                     /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"
     primer_bind     432..451
                     /label=F1ori-R
                     /note="F1 origin, reverse 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"
     regulatory      1572..1581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1578..2468
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     protein_bind    2478..2502
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4713..5301
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4766..4786)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5184..5195)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5325..5446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5336..5355)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5390..5409
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5608..5737
                     /label=AAV2 ITR
     rep_origin      complement(5974..6562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6054..6073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6733..7524,7525..7593))
                     /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     7356..7375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7594..7641,1..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 accatggatg
     1501 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
     1561 tcgcaccggt cgccaccatg gcagaaatcg gtactggctt tccattcgac ccccattatg
     1621 tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg
     1681 tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg
     1741 ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac
     1801 cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc
     1861 tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact
     1921 gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta
     1981 tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca
     2041 ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga
     2101 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     2161 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     2221 cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc
     2281 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc
     2341 tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc
     2401 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtcg acgctcgaga
     2461 tttccggctc cggatcaaca agtttgtaca aaaaagcagg cttaacctgc gagcttgtgc
     2521 cccggcacct ccgcgggcgg cgggcgactg gctctgccgc aactgccgcc tcctctctcg
     2581 ccgcggcggc cggcgatagc ccggcgctca tgacagatcc ctgcatgtca ctgagtccac
     2641 catgctttac agaagaagac agatttagtc tggaagctct tcaaacaata cataaacaaa
     2701 tggatgatga caaagatggt ggaattgaag tagaggaaag tgatgaattc atcagagaag
     2761 atatgaaata taaagatgct actaataaac acagccatct gcacagagaa gataaacata
     2821 taacgattga ggatttatgg aaacgatgga aaacatcaga agttcataat tggacccttg
     2881 aagacactct tcagtggttg atagagtttg ttgaactacc ccaatatgag aagaatttta
     2941 gagacaacaa tgtcaaagga acgacacttc ccaggatagc agtgcacgaa ccttcattta
     3001 tgatctccca gttgaaaatc agtgaccgga gtcacagaca aaaacttcag ctcaaggcat
     3061 tggatgtggt tttgtttgga cctctaacac gcccacctca taactggatg aaagatttta
     3121 tcctcacaat ttctatagta attggtgttg gaggctgctg gtttgcttat acgcagaata
     3181 agacatcaaa agaacatgtt gcaaaaatga tgaaagattt agagagctta caaactgcag
     3241 agcaaagtct aatggactta caagagaggc ttgaaaaggc acaggaagaa aacagaaatg
     3301 ttgctgtaga aaagcaaaat ttagagcgca aaatgatgga tgaaatcaat tatgcaaagg
     3361 aggaggcttg tcggctgaga gagctaaggg agggagctga atgtgaattg agtagacgtc
     3421 agtatgcaga acaggaattg gaacaggttc gcatggctct gaaaaaggcc gaaaaagaat
     3481 ttgaactgag aagcagttgg tctgttccag atgcacttca gaaatggctt cagttaacac
     3541 atgaagtaga agtgcaatac tacaatatta aaagacaaaa cgctgaaatg cagctagcta
     3601 ttgctaaaga tgaggcagaa aaaattaaaa agaagagaag cacagtcttt gggactctgc
     3661 acgttgcaca cagctcctcc ctagatgagg tagaccacaa aattctggaa gcaaagaaag
     3721 ctctctctga gttgacaact tgtttacgag aacgactttt tcgctggcaa caaattgaga
     3781 agatctgtgg ctttcagata gcccataact caggactccc cagcctgacc tcttcccttt
     3841 attctgatca cagctgggtg gtgatgccca gagtctccat tccaccctat ccaattgctg
     3901 gaggagttga tgacttagat gaagacacac ccccaatagt gtcacaattt cccgggacca
     3961 tggctaaacc tcctggatca ttagccagaa gcagcagcct gtgccgttca cgccgcagca
     4021 ttgtgccgtc ctcgcctcag cctcagcgag ctcagcttgc tccacacgcc ccccacccgt
     4081 cacaccctcg gcaccctcac cacccgcaac acacaccaca ctccttgcct tcccctgatc
     4141 cagatatcct ctcagtgtca agttgccctg cgctttatcg aaatgaagtg gaggaagagg
     4201 ccatttactt ctctgctgaa aagcaatggg aagtgccaga cacagcttca gaatgtgact
     4261 ccttaaattc ttccattgga aggaaacagt ctcctccttt aagcctcgag atataccaaa
     4321 cattatctcc gcgaaagata tcaagagatg aggtgtccct agaggattcc tcccgagggg
     4381 attcgcctgt aactgtggat gtgtcttggg gttctcccga ctgtgtaggt ctgacagaga
     4441 ctaagagtat gatcttcagt cctgcaagca aagtgtacaa tggcattttg gagaaatcct
     4501 gtagcatgaa ccagctttcc agtggcatcc cggtgcctaa acctcgccac acatcatgtt
     4561 cctcagctgg caacgacagt aaaccagttc aggaagcccc aagtgttgcc agaataagca
     4621 gcatcccaca tgacctttgt cataatggag agaaaagcaa aaagccatca aaaatcaaaa
     4681 gcctttttaa gaagaaatct aagtgaaagc ttaatcaacc tctggattac aaaatttgtg
     4741 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     4801 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4861 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4921 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4981 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5041 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5101 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5161 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5221 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5281 ccctttgggc cgcctccccg cgcggccgct tcgagcagac atgataagat acattgatga
     5341 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     5401 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     5461 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     5521 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt
     5581 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     5641 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     5701 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg
     5761 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5821 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6721 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6781 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6841 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6901 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6961 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7021 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7081 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7141 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7201 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7261 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7321 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7381 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7441 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7501 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7561 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7621 catttatcag ggttattgtc t
//
