LOCUS       sequence-327735-        5580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing CaMPARI2_F391W primarily in excitatory neurons
            in rodent cortex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  AUTHORS   Trojanowski NF, Bottorff J, Turrigiano GG
  TITLE     Activity labeling in vivo using CaMPARI2 reveals intrinsic and 
            synaptic differences between neurons with high and low firing rate 
            set points.
  JOURNAL   Neuron. 2021 Feb 17;109(4):663-676.e5. doi: 
            10.1016/j.neuron.2020.11.027. Epub 2020 Dec 16.
  PUBMED    33333001
REFERENCE   2  (bases 1 to 5580)
  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"; doi: 
            "10.1016/j.neuron.2020.11.027"; journalName: "Neuron"; date: 
            "2021-02-17- 17"; volume: "109"; issue: "4"; pages: "663-676.e5"
COMMENT     Sequence Label: pAAV_CaMKII_NES-his-CaMPARI2-F391W-WPRE-SV40
FEATURES             Location/Qualifiers
     source          1..5580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(142..161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(821..1612,1613..1681))
                     /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     1444..1463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1682..1786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1854..1872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1910..1932)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2032..2051
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2068..2523)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2205..2226)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2417..2436
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2598..2738
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2609..2738
                     /label=AAV2 ITR
     CDS             3191..3208
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3215..3655
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             4451..4474
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4493..4519
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4538..4567
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    4589..5177
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4642..4662)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5060..5071)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5180..5196)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5181..5197)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5226..5347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5237..5256)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5291..5310
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5451..5580
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
       61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1801 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1861 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1921 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1981 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2041 agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg
     2101 cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc
     2161 cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga
     2221 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2281 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
     2341 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2401 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg
     2461 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2521 cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2581 ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     2641 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     2701 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcactt gtggactaag
     2761 tttgttcaca tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac
     2821 ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag
     2881 caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
     2941 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
     3001 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt
     3061 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc ggcaaggatc
     3121 cgccaccatg ctgcagaacg agcttgctct taagttggct ggacttgata ttaacaagac
     3181 tggaggttct catcatcacc accaccatgg atccgaccaa ctgactgaag agcagatcgc
     3241 agaatttaaa gaggctttct ccctatttga caaggacggg gatgggacaa taacaaccaa
     3301 ggagctgggg acggtgatgc ggtctctggg gcagaacccc acagaagcag agctgcagga
     3361 catgatcaat gaagtagatg ccgacggtga cggcacaatc gacttccctg agttcctgac
     3421 aatgatggca agaaaaatga aagacacaga cagtgaagaa gaaattagag aagcgttccg
     3481 tgtgtttgat aaggatggca atggctacat cagtgcagca gagcttcgcc acgtgatgac
     3541 aaaccttgga gagaagttaa cagatgaaga ggttgatgaa atgatcaggg aagcagacat
     3601 cgatggggat ggtcaggtaa actacgaaga gtttgtagtg atgatgacag cgaagctcga
     3661 gtgcgagaaa atctatgtgc gtgatggagt gctgacgggt gatattcata tgatcttgtt
     3721 gcttgaagga aatgcccatt accgatgtga cttcagaact acttacaaag ctaaggagaa
     3781 gggtgtcaag ttaccaggcg tgcactatgt ggaccacacc attgagattt taagccatga
     3841 caaagattac aacaaggtta agatctatga gtatgctgtt gctcattctg gattgcctga
     3901 caatgccaga cgaggcggta ccggcggatc catggtgagt gcgattaagc cagacatgaa
     3961 gatcaaactg cgtatggaag gcaacgtaaa cgggcaccac tttgtgatcg acggagatgg
     4021 tacaggcaag ccttatgagg gaaaacagac catggatctt gaagtcaaag agggcggacc
     4081 tctgcctttt gcctttgata tcctgaccac tgcattccat tacggcaaca gggtattcgt
     4141 gaaatatcca gacaacatac aagactattt taagcagtcg tttcctaagg ggtattcgtg
     4201 ggaacgaagc atgactttcg aagacggggg catttgctat gccagaagcg acataacaat
     4261 ggaaggggac actttctata ataaagttcg attttatggt accaactttc ccgccaatgg
     4321 tccagttatg cagaagaaga cgctgaaatg gatgccgagc tggacgcgtt catcacgtcg
     4381 taagttcaat aagacaggtc acgcactgag agctataggt cggctgagct caggaggttc
     4441 tggaggttct gattacaagg acgacgatga caagggaggt tctggaggtt cttacccata
     4501 cgacgtacca gattacgctg gaggttctgg aggttctgag cagaagctga tctcagagga
     4561 ggacctgcgt acgtaaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     4621 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4681 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4741 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4801 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4861 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4921 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4981 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5041 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5101 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5161 ttgggccgcc tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga
     5221 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     5281 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     5341 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     5401 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atgcggccgc aggaacccct
     5461 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     5521 aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
//
