LOCUS       sequence-338900-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transfer plasmid for BRIC (Bioluminescent Red Indicator for 
            Calcium).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5987)
  AUTHORS   Tian X, Zhang Y, Li X, Xiong Y, Wu T, Ai HW
  TITLE     A luciferase prosubstrate and a red bioluminescent calcium indicator
            for imaging neuronal activity in mice.
  JOURNAL   Nat Commun. 2022 Jul 8;13(1):3967. doi: 10.1038/s41467-022-31673-x.
  PUBMED    35803917
REFERENCE   2  (bases 1 to 5987)
  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"; journalName: "Nat 
            Commun."; date: "2022-07-8"; volume: "13(1):3967. doi"; pages: " 
            10.1038/s41467-022-31673-x"
COMMENT     Sequence Label: pAAV-hSyn-BRIC
FEATURES             Location/Qualifiers
     source          1..5987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   76..205
                     /label=AAV2 ITR
     promoter        281..728
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      757..766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(896..914)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1164..1183
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2287..2364
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     misc_feature    2500..3088
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2553..2573)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2971..2982)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3091..3107)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3092..3108)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3137..3258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3202..3221
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3420..3549
                     /label=AAV2 ITR
     rep_origin      complement(3786..4374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3866..3885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4545..5336,5337..5405))
                     /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     5168..5187
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5406..5510)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5536..5987,1..4))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5673..5694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5885..5904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
       61 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
      121 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      181 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
      241 acgtagccat gctctaggaa gatccagggc cctgcgtatg agtgcaagtg ggttttagga
      301 ccaggatgag gcggggtggg ggtgcctacc tgacgaccgg ccccgaccca ctggacaagc
      361 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      421 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      481 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      541 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      601 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      661 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      721 gagcgcagtc caattcaagc tgctagcaag gatcccgcca ccatggtgag caagggcgag
      781 gcagtgatca aggagttcat gcggttcaag gtgcacatgg agggctccat gaacggccac
      841 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
      901 ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg acatcctgtc ccctcagttc
      961 atgtacggct ccagggcctt catcaagcac cccgccgaca tccccgacta ctataagcag
     1021 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgccgtg
     1081 accgtgaccc aggacacctc cctggaggac ggcaccctga tctacaaggt gaagctccgc
     1141 ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg ctgggaagcg
     1201 tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg gcgacattaa gatggccctg
     1261 cgcctgaagg acggcggccg ctacctggcg gacttcaaga ccatctacaa ggccaagaag
     1321 cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt tggacatcac ctcccacaac
     1381 gaggactaca ccgtggtgga acagtacgaa cgctccgagg gccgccatct tgatacactc
     1441 gaagatttcg ttggggactg gcgacagaca gccggctaca acttgagtca aatccttgaa
     1501 cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg
     1561 attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa
     1621 ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg
     1681 gataatcatc attttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg
     1741 ccgaacttga tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa
     1801 aagatcactg taacagggac cctgattatg catgaccaac tgacagaaga gcagattgca
     1861 gagttcaaag aagccttctc attattcgac aaggatgggg acggcaccat caccacaaag
     1921 gaacttggta ccgttatgag gtcgcttgga caaaacccaa cggaagcaga attgcaggat
     1981 atgatcaatg aagtcgatgc tgatggcaat ggaacgattt actttcctga atttcttact
     2041 atgatggcta gaaaaatgaa ggacacagac agcgaagagg aaatccgaga agcattccgt
     2101 gtttttgaca aggatgggaa cggctacatc agcgctgctc aactacgtca cgtcatgaca
     2161 aaccttgggg agaagttaac agatgaagaa gttgatgaaa tgataaggga agcagatatc
     2221 gatggtgatg gccaagtaaa ctatgaagag tttgtacaaa tgatgacagc aaaggggggc
     2281 gagtccaaga ggcgctggaa gaaaaacttc attgccgtca gcgctgccaa ccggttcaag
     2341 aagatctcca gctccggggc actggagctc tggaaaggca acaaaattat cgacgagcgc
     2401 ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg
     2461 cgtctgcatg aacgcattct ggcgtaaaag cttatcgata atcaacctct ggattacaaa
     2521 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2581 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2641 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2701 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2761 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2821 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2881 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     2941 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3001 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3061 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg
     3121 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3181 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3241 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3301 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     3361 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     3421 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3481 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3541 agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3601 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3661 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3721 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3781 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3841 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3901 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3961 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4021 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4081 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4141 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4201 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4261 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4321 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4381 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4441 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4501 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4561 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4621 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4681 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4741 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4801 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4861 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4921 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4981 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5041 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5101 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5161 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5221 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5281 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5341 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5401 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5461 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5521 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5581 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5641 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5701 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5761 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5821 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5881 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5941 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggc
//
