LOCUS       sequence-232115-        5426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing CaMPARI2_F391W-L398V (Kd = 174nM), 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 5426)
  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 5426)
  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-F391W-L398V
FEATURES             Location/Qualifiers
     source          1..5426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   138..267
                     /label=AAV2 ITR
     promoter        349..796
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             893..910
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             917..1357
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             2078..2134
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     misc_feature    2150..2738
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2203..2223)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2621..2632)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2741..2757)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2742..2758)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2787..2908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2798..2817)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2852..2871
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3287..3875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3367..3386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4046..4837,4838..4906))
                     /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     4669..4688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4907..5011)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5037..5426,1..66))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5174..5195)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5386..5405
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
       61 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      121 cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
      181 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
      241 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
      301 ctacgtagcc atgctctagg aagatctctg cagagggccc tgcgtatgag tgcaagtggg
      361 ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact
      421 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
      481 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
      541 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
      601 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
      661 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
      721 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
      781 gtgcctgaga gcgcagtcga attcaagctg ctagcaagga tccgccacca tgctgcagaa
      841 cgagcttgct cttaagttgg ctggacttga tattaacaag actggaggtt ctcatcatca
      901 ccaccaccat ggatccgacc aactgactga agagcagatc gcagaattta aagaggcttt
      961 ctccctattt gacaaggacg gggatgggac aataacaacc aaggagctgg ggacggtgat
     1021 gcggtctctg gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga
     1081 tgccgacggt gacggcacaa tcgacttccc tgagttcctg acaatgatgg caagaaaaat
     1141 gaaagacaca gacagtgaag aagaaattag agaagcgttc cgtgtgtttg ataaggatgg
     1201 caatggctac atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt
     1261 aacagatgaa gaggttgatg aaatgatcag ggaagcagac atcgatgggg atggtcaggt
     1321 aaactacgaa gagtttgtag tgatgatgac agcgaagctc gagtgcgaga aaatctatgt
     1381 gcgtgatgga gtgctgacgg gtgatattca tatgatcttg ttgcttgaag gaaatgccca
     1441 ttaccgatgt gacttcagaa ctacttacaa agctaaggag aagggtgtca agttaccagg
     1501 cgtgcactat gtggaccaca ccattgagat tttaagccat gacaaagatt acaacaaggt
     1561 taagatctat gagtatgctg ttgctcattc tggattgcct gacaatgcca gacgaggcgg
     1621 taccggcgga tccatggtga gtgcgattaa gccagacatg aagatcaaac tgcgtatgga
     1681 aggcaacgta aacgggcacc actttgtgat cgacggagat ggtacaggca agccttatga
     1741 gggaaaacag accatggatc ttgaagtcaa agagggcgga cctctgcctt ttgcctttga
     1801 tatcctgacc actgcattcc attacggcaa cagggtattc gtgaaatatc cagacaacat
     1861 acaagactat tttaagcagt cgtttcctaa ggggtattcg tgggaacgaa gcatgacttt
     1921 cgaagacggg ggcatttgct atgccagaag cgacataaca atggaagggg acactttcta
     1981 taataaagtt cgattttatg gtaccaactt tcccgccaat ggtccagtta tgcagaagaa
     2041 gacgctgaaa tggatgccga gctggacgcg ttcatcacgt cgtaagtgga ataagacagg
     2101 tcacgcagtc agagctatag gtcggctgag ctcataaaag cttatcgata atcaacctct
     2161 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2221 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2281 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2341 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2401 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2461 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2521 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2581 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2641 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2701 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc
     2761 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2821 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2881 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2941 ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     3001 aatcgataag gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca
     3061 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     3121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     5041 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5101 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5161 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5221 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5281 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5341 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5401 aggagcgggc gctagggcgc tggcaa
//
