LOCUS       sequence-471085-        7054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of mouse STIM1 internally tagged with mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  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 7054)
  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 mEmerald-STIM1 WPRE
FEATURES             Location/Qualifiers
     source          1..7054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1689..1818
                     /label=AAV2 ITR
     promoter        1900..2347
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          2369..2501
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2484..2503
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2515..2524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2596..2598,2599..2601,2602..3312)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2641..2662)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2902..2921)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3249..3270
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5329..5917
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5382..5402)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5800..5811)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5920..5936)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5921..5937)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5966..6087
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5977..5996)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6031..6050
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6466..7054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6546..6565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     1741 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     1801 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
     1861 catgctctag gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac
     1921 caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca
     1981 cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat
     2041 gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc
     2101 tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc
     2161 cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg
     2221 accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc
     2281 cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag
     2341 agcgcagtcg aattcaagct gctagcaggt aagtatcaag gttacaagac aggtttaagg
     2401 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     2461 tattggtctt actgacatcc actttgcctt tctctccaca ggtgataagg atccgccacc
     2521 atggatgtgt gcgcccgtct tgccctgtgg cttctttggg ggctccttct gcatcagggc
     2581 cagagtggcg gctccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2641 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2701 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2761 gtgccctggc ccaccctcgt gaccaccttg acctacggcg tgcagtgctt cgcccgctac
     2821 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2881 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2941 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3001 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaaggtcta tatcaccgcc
     3061 gacaagcaga agaacggcat caaggtgaac ttcaagaccc gccacaacat cgaggacggc
     3121 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3181 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     3241 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3301 gagctctaca agggcggctc cctcagccat agtcacagtg aaaagaatac aggagcgagc
     3361 tccggggcga cttctgaaga gtctaccgaa gcagagtttt gccgaattga caagcccctg
     3421 tgccacagtg aggatgagaa gctcagcttt gaggccgtcc gaaacatcca taagctgatg
     3481 gatgacgatg ccaatggtga tgtggatgtg gaagaaagtg atgagttcct aagggaagac
     3541 ctcaattacc atgacccaac agtgaaacat agcaccttcc atggtgagga taaacttatc
     3601 agcgtggagg acctgtggaa ggcgtggaag tcatcagaag tgtacaactg gactgtggat
     3661 gaggtgatac agtggctcat tacgtatgtg gagctgccac agtatgagga aaccttccgg
     3721 aagttgcagc ttactggcca cgccatgcca aggcttgcag taaccaacac caccatgaca
     3781 gggactgtac tgaagatgac agatcggagc cacaggcaga agctgcagct gaaggccctg
     3841 gacacagtgc tgtttgggcc tcctctcttg actcggcata atcacctgaa ggacttcatg
     3901 ctggtggtgt ctatcgttat tggtgtgggt ggctgctggt ttgcctatat ccagaaccgt
     3961 tactctaagg agcacatgaa gaaaatgatg aaggatctgg aagggttaca ccgggctgag
     4021 cagagtctgc atgaccttca ggaaaggctg cacaaggccc aggaggagca ccgaactgtg
     4081 gaagtagaga aggtccacct ggagaagaag ctgcgagatg agatcaacct tgccaagcag
     4141 gaagctcagc ggctgaagga gctgagggag ggtactgaga atgagaggag ccgtcaaaaa
     4201 tatgctgagg aagagctgga gcaggttcgg gaggccttga ggaaagcaga gaaggagctg
     4261 gaatcacaca gctcatggta tgctcctgag gccctgcaga agtggctgca gctgacccat
     4321 gaggtggagg tgcagtacta caacatcaag aagcaaaatg cagagaggca gctgctggtg
     4381 gccaaggagg gggctgagaa aataaaaaag aagagaaaca cgctttttgg taccttccat
     4441 gtggcccaca gctcttccct ggatgatgtg gatcataaaa tcctaactgc taagcaagct
     4501 ctgagtgagg tgacagcggc actgagggag cgcctgcacc gatggcagca gatcgagatc
     4561 ctctgcggtt tccagattgt caataacccc ggcatccact ccttggtggc tgctctcaac
     4621 atcgacccca gctggatggg cagcacccgc cctaaccccg cccacttcat catgactgac
     4681 gatgtggatg acatggatga ggagattgtg tcgcccttgt ccatgcagtc ccccagcctg
     4741 cagagcagtg tccggcagcg cctgacggag ccacagcttg gcctgggatc tcagagggat
     4801 ttgacccatt ccgattcgga gtcctccctc cacatgagtg accgccagcg tgtggccccc
     4861 aagcctcctc agatgggccg tgctgcagat gaagctctca atgccatgcc ttccaatggc
     4921 agccatcggc tgattgaggg ggtccatcca ggatctctgg tggagaaact gcctgacagc
     4981 cctgctctgg ccaagaagac atttatggcg ttgaaccatg gcctagacaa ggcccacagc
     5041 ctgatggagc tgaacccctc agtcccacct ggtggctccc cacttttgga ttcttcccat
     5101 tctcttagcc ccagttcccc agacccagac acgccatctc cagttgggga caaccgagct
     5161 ctgcagggta gccgaaacac acgaataccc cacttggctg gcaagaaggc aatggctgag
     5221 gaggataatg gttccattgg tgaggagaca gactccagtc caggcaggaa gaagtttcct
     5281 ctcaaaattt ttaagaagcc tcttaagaag tagtaaaagc ttatcgataa tcaacctctg
     5341 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5401 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5461 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5521 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5581 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5641 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5701 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5761 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5821 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5881 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgaccc
     5941 gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     6001 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     6061 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     6121 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     6181 atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     6241 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     6301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7021 gcagcagatt acgcgcagaa aaaaaggatc tcaa
//
