LOCUS       sequence-264965-        3734 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of GCaMP6M in cells expressing both 
            Cre AND Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3734)
  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 3734)
  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-GCaMP6M
FEATURES             Location/Qualifiers
     source          1..3734
                     /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(1481..1502)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(1715..1771)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     regulatory      1783..1792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1797..1830
                     /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    1881..1914
                     /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    1921..1954
                     /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     2606..2625
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2865..2886
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2994..3027)
                     /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)"
     misc_feature    3136..3724
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3189..3209)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3607..3618)
                     /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 gaagttatgt cgaccctcct gtactcacct tgctcaccat gctccctccg gtaccgccct
     1441 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     1501 tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga ctgcacgctc aggtagtggt
     1561 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtaggcga
     1621 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttcgccttg atgccgttct
     1681 tctgcttgtc ggccttgata tagacgttct cgagtgagct cagccgacct atagctctga
     1741 ctgcgtgacc tgtcttattc cacttacgac gtgatgagtc caccatggtg gctagcataa
     1801 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
     1861 tctttagata ggtgcaaaga ataacttcgt ataaggtata ctatacgaag ttatggtacc
     1921 gaagttccta ttcttcaaat agtataggaa cttcaattgt tattggtgca aagaatcact
     1981 tccttaagtc tttatactgc ccaggaagtt cctattcttc aaaaggtata ggaacttcgg
     2041 cgcgcctcac ttcgctgtca tcatttgtac aaactcttcg tagtttacct gaccatcccc
     2101 atcgatgtct gcttccctga tcatttcatc aacctcttca tctgttaact tctctccaag
     2161 gtttgtcatc acgtggcgaa gctctgctgc actgatgtag ccattgccat ccttatcaaa
     2221 cacaccgaac gcttctctaa tttcttcttc cgtgtccctg tagctccctt ttcttgccat
     2281 cattgtcagg aactcaggga agtcgattgt gccgtcaccg tcggcatcta cttcattgat
     2341 catgtcctgc agctctgctt ctgtggggtt ctgccccaga gaccgcatca ccgtccccag
     2401 ctccttggtt gttattgtcc catccccgtc cttgtcaaat agggagaaag cctctttaaa
     2461 ttctgcgatc tgctcttcag tcagttggtc cggcaggttg tactccagct tgtgccccag
     2521 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     2581 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
     2641 gcgctcctgg atgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     2701 gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca
     2761 gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc
     2821 atcgccctca ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     2881 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc tgtgggacga catgacttaa
     2941 ccagttaacc atgttctctg aacatgagaa gggctggcca ctctccatct agagaagttc
     3001 ctatactatt tgaagaatag gaacttcttg ccttaaccca gaaattatca ctgttattct
     3061 ttagaatagt gcaaagagaa gttcctatac cttttgaaga ataggaactt cgaattcgat
     3121 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3181 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3241 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3301 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3361 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3421 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3481 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3541 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3601 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3661 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3721 ccgcatcgat accg
//
