LOCUS       sequence-497322-        6794 bp    DNA     circular SYN 14-JAN-2026
DEFINITION  An AAV construct designed to express a CRY2-fused truncated STIM1 
            fragment (a.a.318–450) under the control of the GfaABC1D promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6794)
  AUTHORS   Kook YH, Lee H, Lee J, Jeong Y, Rho J, Heo WD, Lee S
  TITLE     AAV-compatible optogenetic tools for activating endogenous calcium 
            channels in vivo.
  JOURNAL   Mol Brain. 2023 Oct 17;16(1):73. doi: 10.1186/s13041-023-01061-7.
  PUBMED    37848907
REFERENCE   2  (bases 1 to 6794)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 14, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol 
            Brain."; date: "2023-10-17"; volume: "16(1):73. doi"; pages: " 
            10.1186/s13041-023-01061-7"
COMMENT     Sequence Label: pAAV-GfaABC1D-EGFP-CRY2-STIM1(318-450)
FEATURES             Location/Qualifiers
     source          1..6794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      857..866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(863..865,866..868,869..1579)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(863..865,866..868,869..1579)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(908..929)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1169..1188)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1516..1537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3657..3677)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4057..4197
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4057..4186
                     /label=AAV2 ITR
     rep_origin      4272..4727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4359..4378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4569..4590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4744..4763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4863..4885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4923..4941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5009..5113
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5114..5182,5183..5974)
                     /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"
     rep_origin      6145..6733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6634..6653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacata tcctggtgtg gagtagggga cgctgctctg
      181 acagaggctc gggggcctga gctggctctg tgagctgggg aggaggcaga cagccaggcc
      241 ttgtctgcaa gcagacctgg cagcattggg ctggccgccc cccagggcct cctcttcatg
      301 cccagtgaat gactcacctt ggcacagaca caatgttcgg ggtgggcaca gtgcctgctt
      361 cccgccgcac cccagccccc ctcaaatgcc ttccgagaag cccattgagc agggggcttg
      421 cattgcaccc cagcctgaca gcctggcatc ttgggataaa agcagcacag ccccctaggg
      481 gctgcccttg ctgtgtggcg ccaccggcgg tggagaacaa ggctctattc agcctgtgcc
      541 caggaaaggg gatcagggga tgcccaggca tggacagtgg gtggcagggg gggagaggag
      601 ggctgtctgc ttcccagaag tccaaggaca caaatgggtg aggggagagc tctccccata
      661 gctgggctgc ggcccaaccc caccccctca ggctatgcca gggggtgttg ccaggggcac
      721 ccgggcatcg ccagtctagc ccactccttc acaaagccct cgcatcccag gagcgagcag
      781 agccagagca ggttggagag gagacgcatc acctccgctg ctcgctctag agccgatccc
      841 cgggtaccgg gctagcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1021 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1441 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1561 catggacgag ctgtacaagc caccggtcat gaagatggac aaaaagacca tcgtctggtt
     1621 tcggagagat ttgagaatag aagataatcc cgcgctcgcc gccgcggccc acgagggttc
     1681 cgtcttcccc gttttcattt ggtgtcctga agaagaaggc cagttttatc ccggaagggc
     1741 ctctaggtgg tggatgaagc aaagtctggc ccatcttagc cagtcactga aagcactggg
     1801 cagtgatctt accctgatca agacacacaa taccatctct gccatcctcg actgcatcag
     1861 ggtgaccggc gcaacgaaag tcgtgtttaa ccacctgtac gatccagtta gtctggtgcg
     1921 cgaccacact gtgaaggaga agctggtgga acgggggatc agtgtgcaga gctacaacgg
     1981 ggaccttctg tacgagccat gggagatcta ttgcgagaaa gggaaaccgt tcacctcctt
     2041 caacagttac tggaagaaat gtttggatat gtcaatagag tccgttatgt tgccccctcc
     2101 ctggagactg atgccgatta ctgctgctgc agaggccatc tgggcctgct ccatcgagga
     2161 actcggtctg gaaaatgaag cagaaaagcc aagcaatgca cttctcacta gagcctggag
     2221 ccccggctgg tctaatgccg acaagctgct taacgagttc atcgaaaaac aactgattga
     2281 ctacgcgaag aactccaaga aagtggtagg taactcaact agcttgctct ctccatatct
     2341 ccattttggc gagatttctg tccgccatgt atttcagtgc gctcggatga aacagattat
     2401 ctgggctcgc gataaaaaca gcgaaggcgc cgaaagcgcc gatctgttcc tgcgagggat
     2461 cggacttcgg gaatactccc ggtatatatg tttcaacttt ccattcacac acgagcagag
     2521 tctgttgtcc cacctcaggt tcttcccctg ggacgccgat gtcgacaaat tcaaggcatg
     2581 gagacaggga aggacagggt acccactcgt ggatgctggc atgagagagc tctgggctac
     2641 aggctggatg cacaaccgca tccgggtaat cgtgtcctca tttgctgtca agtttctgct
     2701 cctgccttgg aaatggggaa tgaagtactt ttgggatacc cttctcgacg ccgacttgga
     2761 gtgtgacatt ctgggatggc aatatattag cgggtcaatt cctgacggcc atgagttgga
     2821 caggttggac aatccggcct tgcagggagc taagtatgat cccgaaggag agtatattcg
     2881 acagtggctc cccgagctgg cccgacttcc tacggagtgg attcaccatc cttgggacgc
     2941 accactgaca gtgctcaagg caagcggggt ggagctgggc accaattacg ctaagcctat
     3001 agttgatata gatacagcac gcgagctgct ggctaaagcg atctctcgca ctcgggaggc
     3061 gcagattatg atcggtgctg ccgccaggga cccccccgac ctcgacaacg aattcagtgg
     3121 gggaggcgga gggggtggtt caggtggtgg tggtggttcg ggtggtggcg gagggggagg
     3181 ctccggagag gaggagttgg agcaggttcg ggaggccttg aggaaagcag agaaggagct
     3241 agaatctcac agctcatggt atgctccaga ggcccttcag aagtggctgc agctgacaca
     3301 tgaggtggag gtgcaatatt acaacatcaa gaagcaaaat gctgagaagc agctgctggt
     3361 ggccaaggag ggggctgaga agataaaaaa gaagagaaac acactctttg gcaccttcca
     3421 cgtggcccac agctcttccc tggatgatgt agatcataaa attctaacag ctaagcaagc
     3481 actgagcgag gtgacagcag cattgcggga gcgcctgcac cgctggcaac agatcgagat
     3541 cctctgtggc ttccagattg tcaacaaccc tggcatccac tcactgtagg gatccctcga
     3601 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3661 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3721 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     3781 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3841 tccgtggtgt ttatttgtga aatttgtgat gctattgctt tatttgtaac catctagctt
     3901 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3961 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     4021 tttttaaagc ggccgcagga cggaccgagc ggccgcagga acccctagtg atggagttgg
     4081 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4141 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4201 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4261 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4321 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     4381 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4441 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4501 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4561 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     4621 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     4681 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4741 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4801 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4861 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4921 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4981 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5041 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5101 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5161 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5221 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5281 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5341 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5401 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5461 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5521 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5581 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5641 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5701 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5761 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5821 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5881 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5941 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6001 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6061 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6121 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6181 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6241 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6301 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6361 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6421 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6481 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6541 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6601 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6661 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6721 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6781 ttttgctcac atgt
//
