LOCUS       sequence-199081-        6372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of CaMPARI, a photoconvertible fluorescent 
            protein-based calcium integrator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  AUTHORS   Fosque BF, Sun Y, Dana H, Yang CT, Ohyama T, Tadross MR, Patel R, 
            Zlatic M, Kim DS, Ahrens MB, Jayaraman V, Looger LL, Schreiter ER
  TITLE     Neural circuits. Labeling of active neural circuits in vivo with 
            designed calcium integrators.
  JOURNAL   Science. 2015 Feb 13;347(6223):755-60. doi: 10.1126/science.1260922.
  PUBMED    25678659
REFERENCE   2  (bases 1 to 6372)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2015-02-13- 13"; volume: "347"; 
            issue: "6223"; pages: "755-60"
COMMENT     Sequence Label: pENN.AAV.CAG.Flex.CaMPARI.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   24..153
                     /label=AAV2 ITR
     promoter        464..740
                     /label=chicken beta-actin promoter
     primer_bind     1019..1041
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1103..1122
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1157..1190
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1253..1286
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1399..1416
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1423..1863
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             2584..2640
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     protein_bind    complement(2660..2693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2764..2797)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2839..3427
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2892..2912)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3310..3321)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3470..3591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3481..3500)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3535..3554
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3753..3882
                     /label=AAV2 ITR
     rep_origin      complement(4119..4707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4199..4218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4878..5669,5670..5738))
                     /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     5501..5520
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5739..5843)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5869..6324)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6006..6027)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6218..6237
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
       61 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
      121 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
      181 acttatctac gtagccatgc tctaggaaga gtaccattga cgtcaataat gacgtatgtt
      241 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      301 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      361 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      421 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
      481 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      541 ttttaattat tttgtgcagc gatgggggcg gggggggggg ggggcgcgcg ccaggcgggg
      601 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      661 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      721 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
      781 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
      841 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
      901 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
      961 gcggctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1021 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     1081 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     1141 aaagaattgg atccccataa cttcgtataa agtatcctat acgaagttat atcaaaatag
     1201 gaagaccaat gcttcaccat cgacccgaat tgccaagcat caccatcgac ccataacttc
     1261 gtataatgta tgctatacga agttatacta gggatccccc ctcgaaattc aagctgctag
     1321 caaggatccg ccaccatgct gcagaacgag cttgctctta agttggctgg acttgatatt
     1381 aacaagactg gaggttctca tcatcaccac caccatggat ccgaccaact gactgaagag
     1441 cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacaata
     1501 acaaccaagg agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag
     1561 ctgcaggaca tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag
     1621 ttcctgacaa tgatggcaag aaaaatgaaa gacacagaca gtgaagaaga aattagagaa
     1681 gcgttccgtg tgtttgataa ggatggcaat ggctacatca gtgcagcaga gcttcgccac
     1741 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa
     1801 gcagacatcg atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
     1861 aagctcgagt gcgagaaaat ctatgtgcgt gatggagtgc tgacgggtga tattcatatg
     1921 atcttgttgc ttgaaggaaa tgcccattac cgatgtgact tcagaactac ttacaaagct
     1981 aaggagaagg gtgtcaagtt accaggcgtg cactttgtgg accactgcat tgagatttta
     2041 agccatgaca aagattacaa caaggttaag ctgtatgagt atgctgttgc tcattctgga
     2101 ttgcctgaca atgccagacg aggcggtacc ggcggatcca tggtgagtgc gattaagcca
     2161 gacatgaaga tcaaactccg tatggaaggc aacgtaaacg ggcaccactt tgtgatcgac
     2221 ggagatggta caggcaagcc ttatgaggga aaacagacca tggatcttga agtcaaagag
     2281 ggcggacctc tgccttttgc ctttgatatc ctgaccactg cattccatta cggcaacagg
     2341 gtattcgtga aatatccaga caacatacaa gactatttta agcagtcgtt tcctaagggg
     2401 tattcgtggg aacgaagcat gactttcgaa gacgggggca tttgctatgc cagaaacgac
     2461 ataacaatgg aaggggacac tttctataat aaagttcgat tttatggtac caactttccc
     2521 gccaatggtc cagttatgca gaagaagacg ctgaaatgga tgccgagctg gacgcgttca
     2581 tcacgtcgta agtggaataa gacaggtcac gcagtcagag ctataggtcg gctgagctca
     2641 taaaagctcg aattctagta taacttcgta taggatactt tatacgaagt tatcattggg
     2701 attcttccta ttttgatcca agcatcacca tcgaccctct agtccagatc tcaccatcga
     2761 cccataactt cgtatagcat acattatacg aagttatgtc cctcgaagag gttcgaattc
     2821 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2881 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2941 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3001 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3061 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3121 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3181 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3241 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3301 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3361 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3421 tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt
     3481 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3541 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3601 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3661 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     3721 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     3781 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3841 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc
     3901 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3961 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4021 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4081 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4141 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4201 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4261 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4321 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4381 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4441 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4501 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4561 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4621 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4681 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4741 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4801 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4861 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4921 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4981 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5041 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5101 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5161 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5221 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5281 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5341 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5401 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5461 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5521 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5581 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5641 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5701 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5761 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5821 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     5881 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5941 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6001 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6061 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6121 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6181 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6241 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6301 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     6361 ggcgatcggt gc
//
