LOCUS       sequence-194341-        5567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing CaMPARI2 (Kd = 200nM), 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-WPRE-SV40
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        197..644
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             741..758
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             765..1205
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             2001..2024
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2043..2069
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2088..2117
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2139..2727
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2192..2212)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2610..2621)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2730..2746)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2731..2747)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2776..2897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2787..2806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2841..2860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3276..3864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3356..3375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4035..4826,4827..4895))
                     /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     4658..4677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4896..5000)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5026..5481)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5163..5184)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5375..5394
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5553..5567,1..115)
                     /label=AAV2 ITR
ORIGIN
        1 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
       61 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      121 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatctctgca
      181 gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg
      241 cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg
      301 cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc
      361 cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc
      421 tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc
      481 accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga
      541 taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt
      601 ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat tcaagctgct
      661 agcaaggatc cgccaccatg ctgcagaacg agcttgctct taagttggct ggacttgata
      721 ttaacaagac tggaggttct catcatcacc accaccatgg atccgaccaa ctgactgaag
      781 agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg gatgggacaa
      841 taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc acagaagcag
      901 agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacaatc gacttccctg
      961 agttcctgac aatgatggca agaaaaatga aagacacaga cagtgaagaa gaaattagag
     1021 aagcgttccg tgtgtttgat aaggatggca atggctacat cagtgcagca gagcttcgcc
     1081 acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa atgatcaggg
     1141 aagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtagtg atgatgacag
     1201 cgaagctcga gtgcgagaaa atctatgtgc gtgatggagt gctgacgggt gatattcata
     1261 tgatcttgtt gcttgaagga aatgcccatt accgatgtga cttcagaact acttacaaag
     1321 ctaaggagaa gggtgtcaag ttaccaggcg tgcactatgt ggaccacacc attgagattt
     1381 taagccatga caaagattac aacaaggtta agatctatga gtatgctgtt gctcattctg
     1441 gattgcctga caatgccaga cgaggcggta ccggcggatc catggtgagt gcgattaagc
     1501 cagacatgaa gatcaaactg cgtatggaag gcaacgtaaa cgggcaccac tttgtgatcg
     1561 acggagatgg tacaggcaag ccttatgagg gaaaacagac catggatctt gaagtcaaag
     1621 agggcggacc tctgcctttt gcctttgata tcctgaccac tgcattccat tacggcaaca
     1681 gggtattcgt gaaatatcca gacaacatac aagactattt taagcagtcg tttcctaagg
     1741 ggtattcgtg ggaacgaagc atgactttcg aagacggggg catttgctat gccagaagcg
     1801 acataacaat ggaaggggac actttctata ataaagttcg attttatggt accaactttc
     1861 ccgccaatgg tccagttatg cagaagaaga cgctgaaatg gatgccgagc tggacgcgtt
     1921 catcacgtcg taagttcaat aagacaggtc acgcactgag agctataggt cggctgagct
     1981 caggaggttc tggaggttct gattacaagg acgacgatga caagggaggt tctggaggtt
     2041 cttacccata cgacgtacca gattacgctg gaggttctgg aggttctgag cagaagctga
     2101 tctcagagga ggacctgcgt acgtaaaagc ttatcgataa tcaacctctg gattacaaaa
     2161 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2221 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2281 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2341 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2401 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2461 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2521 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     2581 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2641 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     2701 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgaccc gggcggccgc
     2761 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2821 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2881 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2941 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     3001 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     3061 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgactcg
     3121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4141 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     5041 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5101 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5161 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5221 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5281 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5341 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5401 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5461 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     5521 gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcg
//
