LOCUS       sequence-237845-        6122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid for expressing rsCaMPARI in neurons, nucleus localized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Sha F, Abdelfattah AS, Patel R, Schreiter ER
  TITLE     Erasable labeling of neuronal activity using a reversible calcium 
            marker.
  JOURNAL   Elife. 2020 Sep 15;9. pii: 57249. doi: 10.7554/eLife.57249.
  PUBMED    32931424
REFERENCE   2  (bases 1 to 6122)
  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: "Elife."; 
            date: "2020-09-15"; volume: "9. pii: 57249. doi"; pages: " 
            10.7554/eLife.57249"
COMMENT     Sequence Label: pAAV-hsyn_NLS-His-rsCaMPARI-mRuby3
FEATURES             Location/Qualifiers
     source          1..6122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   72..201
                     /label=AAV2 ITR
     promoter        283..730
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      758..767
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             770..790
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             797..814
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2057..2764
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     misc_feature    2780..3368
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2833..2853)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3251..3262)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3371..3387)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3372..3388)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3417..3538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3428..3447)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3482..3501
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3917..4505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3997..4016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4676..5467,5468..5536))
                     /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     5299..5318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5537..5641)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5667..6122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5804..5825)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6016..6035
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
      121 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      181 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      241 agccatgctc taggaagatc tctgcagagg gccctgcgta tgagtgcaag tgggttttag
      301 gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa
      361 gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag
      421 gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc
      481 gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg
      541 tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc
      601 cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg
      661 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct
      721 gagagcgcag tcgaattcaa gctgctagca aggatccgcc accatgggtc ccaagaagaa
      781 gaggaaagtc ggttctcatc atcaccacca ccatggatcc atggtgagtg cgattaagcc
      841 agacatgaag atcaaactgc gtatggaagg caacgtaaac gggcaccact ttgtgatcga
      901 cggagatggt acaggcaagc cttatgaggg aaaacagacc atggatcttg aagtcaaaga
      961 gggcggacct ctgccttttg cctttgatat cctgaccact gcattcctgt acggcaacag
     1021 ggtattcgtg aaatatccag acaacataca agactatttt aagcagtcgt ttcctaaggg
     1081 gtattcgtgg gaacgaagca tgactttcga agacgggggc atttgctatg ccagaagcga
     1141 cataacaatg gaaggggaca ctttctataa taaagttcga ttttatggta ccaactttcc
     1201 cgccaatggt ccagttatgc agaagaagac gctgaaatgg gagccctcca ctgagaaaat
     1261 gtatgtgcgt gatggagtgc tgacgggtga tgtagagatg gctttgttgc ttgaaggaaa
     1321 tgcccattac cgagcgttgt caagccgccg caagttcaac aaaaccggcc atgccttgcg
     1381 tgcaattggc cgcctgtcca gcggaggctc tggcggttcc ggcggttcag gtggatcaga
     1441 ccagctgacc gaggagcaaa ttgcagaatt taaggaggcg ttctctttgt ttgacaaaga
     1501 tggcgacggt acaattacaa ccaaggaatt gggcacagta atgcgctccc ttggacagaa
     1561 tcccactgaa gccgaacttc aagacatgat caatgaggtt gatgctgacg gggatgggac
     1621 tatcgacttt cctgagtttc ttaccatgat ggctcgcaaa atgaaggata cagacagtga
     1681 ggaggaaatt cgtgaggcct ttcgtgtgtt tgataaggac gggaacggtt atatctcagc
     1741 ggcggagctg cgtcacgtta tgaccaactt aggcgaaaag ctgactgatg aagaagtcga
     1801 tgagatgatc cgcgaggcgg acatcgatgg tgatggtcaa gtcaattatg aagagtttgt
     1861 ggtgatgatg accgcgaagt gcctcttcag aactacttac aaagctaagg agaagggtgt
     1921 caagttacca ggcgtgcact atgtggacca caccattgag attttaagcc atgacaaaga
     1981 ttacaacaag gttaagatct atgagtatgc tgttgctcat tctggattgc ctgacaatgc
     2041 cagacgagga ggtagcgtgt ctaagggcga agagctgatc aaggaaaata tgcgtatgaa
     2101 ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa tgcacaggtg aaggagaagg
     2161 cagaccgtac gagggagtgc aaaccatgag gatcaaagtc atcgagggag gacccctgcc
     2221 atttgccttt gacattcttg ccacgtcgtt catgtatggc agccgtacct ttatcaagta
     2281 cccggccgac atccctgatt tctttaaaca gtcctttcct gagggtttta cttgggaaag
     2341 agttacgaga tacgaagatg gtggagtcgt caccgtcacg caggacacca gccttgagga
     2401 tggcgagctc gtctacaacg tcaaggtcag aggggtaaac tttccctcca atggtcccgt
     2461 gatgcagaag aagaccaagg gttgggagcc taatacagag atgatgtatc cagcagatgg
     2521 tggtctgaga ggatacactg acatcgcact gaaagttgat ggtggtggcc atctgcactg
     2581 caacttcgtg acaacttaca ggtcaaaaaa gaccgtcggg aacatcaaga tgcccggtgt
     2641 ccatgccgtt gatcaccgcc tggaaaggat cgaggagagt gacaatgaaa cctacgtagt
     2701 gcaaagagaa gtggcagttg ccaaatacag caaccttggt ggtggcatgg acgagctgta
     2761 caagtaaaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2821 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2881 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2941 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3001 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3061 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3121 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3181 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3241 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3301 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3361 ctccccgcat cgataccgtc gacctcgacc cgggcggccg cttcgagcag acatgataag
     3421 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3481 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3541 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     3601 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     3661 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     3721 tggccactcc ctctctgcgc gctcgctcgc tcactgactc gctgcgctcg gtcgttcggc
     3781 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3841 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3901 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3961 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4021 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4081 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4141 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4201 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4261 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4321 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4381 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4441 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4501 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4561 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4621 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4681 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4741 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4801 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4861 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4921 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4981 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5041 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5101 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5161 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5221 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5281 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5341 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5401 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5461 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5521 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5581 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5641 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     5701 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5761 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5821 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5881 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5941 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     6001 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     6061 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     6121 gt
//
