LOCUS       sequence-194371-        5567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing CaMPARI2_L398T (Kd = 825nM), a 
            photoconvertible fluorescent protein-based calcium integrator, in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5567)
  AUTHORS   Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC, 
            Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA, 
            Sachdev RNS, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE, 
            Oertner TG, Hope BT, Schreiter ER
  TITLE     Improved methods for marking active neuron populations.
  JOURNAL   Nat Commun. 2018 Oct 25;9(1):4440. doi: 10.1038/s41467-018-06935-2.
  PUBMED    30361563
REFERENCE   2  (bases 1 to 5567)
  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: "2018-10-25"; volume: "9(1):4440. doi"; pages: " 
            10.1038/s41467-018-06935-2"
COMMENT     Sequence Label: pAAV_hsyn_NES-his-CaMPARI2-L398T-WPRE-SV40
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..456
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             553..570
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             577..1017
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             1813..1836
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1855..1881
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1900..1929
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1951..2539
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2004..2024)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2422..2433)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2542..2558)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2543..2559)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2588..2709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2599..2618)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2653..2672
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3088..3676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3168..3187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3847..4638,4639..4707))
                     /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     4470..4489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4708..4812)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4838..5293)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4975..4996)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5187..5206
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5365..5494
                     /label=AAV2 ITR
ORIGIN
        1 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
       61 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
      121 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
      181 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
      241 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
      301 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
      361 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
      421 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga attcaagctg ctagcaagga
      481 tccgccacca tgctgcagaa cgagcttgct cttaagttgg ctggacttga tattaacaag
      541 actggaggtt ctcatcatca ccaccaccat ggatccgacc aactgactga agagcagatc
      601 gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc
      661 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
      721 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
      781 acaatgatgg caagaaaaat gaaagacaca gacagtgaag aagaaattag agaagcgttc
      841 cgtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
      901 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
      961 atcgatgggg atggtcaggt aaactacgaa gagtttgtag tgatgatgac agcgaagctc
     1021 gagtgcgaga aaatctatgt gcgtgatgga gtgctgacgg gtgatattca tatgatcttg
     1081 ttgcttgaag gaaatgccca ttaccgatgt gacttcagaa ctacttacaa agctaaggag
     1141 aagggtgtca agttaccagg cgtgcactat gtggaccaca ccattgagat tttaagccat
     1201 gacaaagatt acaacaaggt taagatctat gagtatgctg ttgctcattc tggattgcct
     1261 gacaatgcca gacgaggcgg taccggcgga tccatggtga gtgcgattaa gccagacatg
     1321 aagatcaaac tgcgtatgga aggcaacgta aacgggcacc actttgtgat cgacggagat
     1381 ggtacaggca agccttatga gggaaaacag accatggatc ttgaagtcaa agagggcgga
     1441 cctctgcctt ttgcctttga tatcctgacc actgcattcc attacggcaa cagggtattc
     1501 gtgaaatatc cagacaacat acaagactat tttaagcagt cgtttcctaa ggggtattcg
     1561 tgggaacgaa gcatgacttt cgaagacggg ggcatttgct atgccagaag cgacataaca
     1621 atggaagggg acactttcta taataaagtt cgattttatg gtaccaactt tcccgccaat
     1681 ggtccagtta tgcagaagaa gacgctgaaa tggatgccga gctggacgcg ttcatcacgt
     1741 cgtaagttca ataagacagg tcacgcaacg agagctatag gtcggctgag ctcaggaggt
     1801 tctggaggtt ctgattacaa ggacgacgat gacaagggag gttctggagg ttcttaccca
     1861 tacgacgtac cagattacgc tggaggttct ggaggttctg agcagaagct gatctcagag
     1921 gaggacctgc gtacgtaaaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     1981 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2041 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2101 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2161 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2221 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2281 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2341 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2401 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2461 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2521 ctttgggccg cctccccgca tcgataccgt cgacctcgac ccgggcggcc gcttcgagca
     2581 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2641 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2701 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     2761 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct
     2821 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     2881 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgact cgctgcgctc
     2941 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3001 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3061 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3121 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3181 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3241 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3301 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3361 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3421 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3481 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3541 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3601 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3661 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3721 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3781 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3841 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3901 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3961 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4021 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4081 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4141 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4201 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4261 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4321 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4381 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4441 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4501 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4561 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4621 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4681 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4741 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4801 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     4861 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4921 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4981 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5041 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5101 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5161 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5221 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5281 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5341 cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     5401 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     5461 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
     5521 tacttatcta cgtagccatg ctctaggaag atctctgcag agggccc
//
