LOCUS       sequence-471211-        7446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of human STIM2 internally tagged with mScarlet-I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7446)
  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 7446)
  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 mScarlet-STIM2 WPRE
FEATURES             Location/Qualifiers
     source          1..7446
                     /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..2273
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1711..1729)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1979..1998
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    2283..2307
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4518..5106
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4571..4591)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4989..5000)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5130..5251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5141..5160)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5195..5214
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5413..5542
                     /label=AAV2 ITR
     rep_origin      complement(5779..6367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5859..5878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6538..7329,7330..7398))
                     /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     7161..7180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7399..7446,1..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 gtgagcaagg gcgaggcagt gatcaaggag ttcatgcggt
     1621 tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg
     1681 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     1741 tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg gccttcatca
     1801 agcaccccgc cgacatcccc gactactata agcagtcctt ccccgagggc ttcaagtggg
     1861 agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt gacccaggac acctccctgg
     1921 aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct cctgacggcc
     1981 ccgtaatgca gaagaagaca atgggctggg aagcgtccac cgagcggttg taccccgagg
     2041 acggcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc ggccgctacc
     2101 tggcggactt caagaccacc tacaaggcca agaagcccgt gcagatgccc ggcgcctaca
     2161 acgtcgaccg caagttggac atcacctccc acaacgagga ctacaccgtg gtggaacagt
     2221 acgaacgctc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtccggat
     2281 caacaagttt gtacaaaaaa gcaggcttaa cctgcgagct tgtgccccgg cacctccgcg
     2341 ggcggcgggc gactggctct gccgcaactg ccgcctcctc tctcgccgcg gcggccggcg
     2401 atagcccggc gctcatgaca gatccctgca tgtcactgag tccaccatgc tttacagaag
     2461 aagacagatt tagtctggaa gctcttcaaa caatacataa acaaatggat gatgacaaag
     2521 atggtggaat tgaagtagag gaaagtgatg aattcatcag agaagatatg aaatataaag
     2581 atgctactaa taaacacagc catctgcaca gagaagataa acatataacg attgaggatt
     2641 tatggaaacg atggaaaaca tcagaagttc ataattggac ccttgaagac actcttcagt
     2701 ggttgataga gtttgttgaa ctaccccaat atgagaagaa ttttagagac aacaatgtca
     2761 aaggaacgac acttcccagg atagcagtgc acgaaccttc atttatgatc tcccagttga
     2821 aaatcagtga ccggagtcac agacaaaaac ttcagctcaa ggcattggat gtggttttgt
     2881 ttggacctct aacacgccca cctcataact ggatgaaaga ttttatcctc acaatttcta
     2941 tagtaattgg tgttggaggc tgctggtttg cttatacgca gaataagaca tcaaaagaac
     3001 atgttgcaaa aatgatgaaa gatttagaga gcttacaaac tgcagagcaa agtctaatgg
     3061 acttacaaga gaggcttgaa aaggcacagg aagaaaacag aaatgttgct gtagaaaagc
     3121 aaaatttaga gcgcaaaatg atggatgaaa tcaattatgc aaaggaggag gcttgtcggc
     3181 tgagagagct aagggaggga gctgaatgtg aattgagtag acgtcagtat gcagaacagg
     3241 aattggaaca ggttcgcatg gctctgaaaa aggccgaaaa agaatttgaa ctgagaagca
     3301 gttggtctgt tccagatgca cttcagaaat ggcttcagtt aacacatgaa gtagaagtgc
     3361 aatactacaa tattaaaaga caaaacgctg aaatgcagct agctattgct aaagatgagg
     3421 cagaaaaaat taaaaagaag agaagcacag tctttgggac tctgcacgtt gcacacagct
     3481 cctccctaga tgaggtagac cacaaaattc tggaagcaaa gaaagctctc tctgagttga
     3541 caacttgttt acgagaacga ctttttcgct ggcaacaaat tgagaagatc tgtggctttc
     3601 agatagccca taactcagga ctccccagcc tgacctcttc cctttattct gatcacagct
     3661 gggtggtgat gcccagagtc tccattccac cctatccaat tgctggagga gttgatgact
     3721 tagatgaaga cacaccccca atagtgtcac aatttcccgg gaccatggct aaacctcctg
     3781 gatcattagc cagaagcagc agcctgtgcc gttcacgccg cagcattgtg ccgtcctcgc
     3841 ctcagcctca gcgagctcag cttgctccac acgcccccca cccgtcacac cctcggcacc
     3901 ctcaccaccc gcaacacaca ccacactcct tgccttcccc tgatccagat atcctctcag
     3961 tgtcaagttg ccctgcgctt tatcgaaatg aagtggagga agaggccatt tacttctctg
     4021 ctgaaaagca atgggaagtg ccagacacag cttcagaatg tgactcctta aattcttcca
     4081 ttggaaggaa acagtctcct cctttaagcc tcgagatata ccaaacatta tctccgcgaa
     4141 agatatcaag agatgaggtg tccctagagg attcctcccg aggggattcg cctgtaactg
     4201 tggatgtgtc ttggggttct cccgactgtg taggtctgac agagactaag agtatgatct
     4261 tcagtcctgc aagcaaagtg tacaatggca ttttggagaa atcctgtagc atgaaccagc
     4321 tttccagtgg catcccggtg cctaaacctc gccacacatc atgttcctca gctggcaacg
     4381 acagtaaacc agttcaggaa gccccaagtg ttgccagaat aagcagcatc ccacatgacc
     4441 tttgtcataa tggagagaaa agcaaaaagc catcaaaaat caaaagcctt tttaagaaga
     4501 aatctaagtg aaagcttaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4561 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4621 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4681 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4741 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4801 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4861 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4921 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4981 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5041 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5101 ccccgcgcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     5161 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     5221 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     5281 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     5341 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     5401 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5461 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5521 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5581 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6481 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6541 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6601 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6661 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6721 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6781 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6841 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6901 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6961 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7021 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7081 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7141 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7201 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7261 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7321 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7381 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7441 ttgtct
//
