LOCUS       sequence-265011-        4139 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of GCaMP6M in cells expressing Cre 
            AND Flp AND Vcre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4139)
  AUTHORS   Fenno LE, Ramakrishnan C, Kim YS, Evans KE, Lo M, Vesuna S, Inoue M,
            Cheung KYM, Yuen E, Pichamoorthy N, Hong ASO, Deisseroth K
  TITLE     Comprehensive Dual- and Triple-Feature Intersectional Single-Vector 
            Delivery of Diverse Functional Payloads to Cells of Behaving 
            Mammals.
  JOURNAL   Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003.
  PUBMED    32574559
REFERENCE   2  (bases 1 to 4139)
  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: "Neuron. 
            2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003."
COMMENT     Sequence Label: pAAV-Ef1a-Con/Fon/Von GCaMP 6m
FEATURES             Location/Qualifiers
     source          1..4139
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        87..1264
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1255
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1212..1232
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1271..1304)
                     /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(1355..1388)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     primer_bind     complement(1484..1505)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(1718..1774)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     regulatory      1786..1795
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1802..1835
                     /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    1886..1919
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    2015..2048
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     2392..2413
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2469..2502)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     3335..3354
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3541..4129
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3594..3614)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4012..4023)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccactggctg cagtacgtga ttcttgatcc
      421 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      481 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      541 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      601 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      661 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      721 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      781 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
      841 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
      901 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
      961 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1021 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1081 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1141 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1201 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1261 gtgaggatcc ataacttcgt ataggatact ttatacgaag ttatgtagct ggagcaattt
     1321 gaaaagattg ttgtattacc tctagctgcg cttaataact tcgtatagta taccttatac
     1381 gaagttatgg cgcgccaacc ccgacactta ccttgctcac catgctccct ccggtaccgc
     1441 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     1501 ccatgtgatc gcgcttctcg ttggggtctt tcgaaagttt ggactgcacg ctcaggtagt
     1561 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtagg
     1621 cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttcgcc ttgatgccgt
     1681 tcttctgctt gtcggccttg atatagacgt tctcgagtga gctcagccga cctatagctc
     1741 tgactgcgtg acctgtctta ttccacttac gacgtgatga gtccaccatg gtggcaccgg
     1801 tataacttcg tataaagtat cctatacgaa gttatttgcc ttagttattc ttagatagac
     1861 ccagaaatta tcacttgtgc aaagaataac ttcgtataag gtatactata cgaagttatg
     1921 agctctgtgc cccccctttt ttttataaaa ttgtattaat gttatataca tatctcctgt
     1981 atgtgaccca tgtgcttatg actctattaa ttaagaagtt cctattcttc aaatagtata
     2041 ggaacttcgc agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaga
     2101 agttcctatt cttcaaaagg tataggaact tcactagtcc tcctgtactc accttgaaga
     2161 agatggtgcg ctcttggatg tagccctcgg gcatggcgga cttgaagaag tcgtgctgct
     2221 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     2281 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     2341 aggtggcatc gccctcaccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     2401 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgccctaa acacatgaga
     2461 aagtcgacga agttcctata ctatttgaag aataggaact tcttgcctta acccagaaat
     2521 tatcactgtt attctttaga atggtgcaaa gagaagttcc tatacctttt gaagaatagg
     2581 aacttcgcta gctggagagt ggccagccct tctcatgttc agagaacatg gttaactggt
     2641 taagtcatgt ctcaatttct gagaactgtc attctcggaa attgagtagc tggttgaaga
     2701 ttgctctagt attacagcaa tttgaactgc gcttatcaat ttctgagaag tgtctttctc
     2761 ggaaattgag gtacctcact tcgctgtcat catttgtaca aactcttcgt agtttacctg
     2821 accatcccca tcgatgtctg cttccctgat catttcatca acctcttcat ctgttaactt
     2881 ctctccaagg tttgtcatca cgtggcgaag ctctgctgca ctgatgtagc cattgccatc
     2941 cttatcaaac acaccgaacg cttctctaat ttcttcttcc gtgtccctgt agctcccttt
     3001 tcttgccatc attgtcagga actcagggaa gtcgattgtg ccgtcaccgt cggcatctac
     3061 ttcattgatc atgtcctgca gctctgcttc tgtggggttc tgccccagag accgcatcac
     3121 cgtccccagc tccttggttg ttattgtccc atccccgtcc ttgtcaaata gggagaaagc
     3181 ctctttaaat tctgcgatct gctcttcagt cagttggtcc ggcaggttgt actccagctt
     3241 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     3301 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtcctgtgg
     3361 gatctagagt caagattgct ccagcaatct tgtaatactc aatttccgag aatgacagtt
     3421 ctcagaaatt gataagcaga caatcttgta atacatgcag ctagtcaaga ttgctccagc
     3481 tatcaatttc cgagaaagac acttctcaga aattgagaat tcgatatcaa gcttatcgat
     3541 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3601 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3661 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3721 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3781 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3841 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3901 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3961 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4021 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4081 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccg
//
