LOCUS       sequence-264967-        3736 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of GCaMP6M in cells expressing Flp 
            AND NOT Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3736)
  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 3736)
  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-Coff/Fon-GCaMP6M
FEATURES             Location/Qualifiers
     source          1..3736
                     /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."
     regulatory      1395..1404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1416..1472
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1685..1706
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1799..1832
                     /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    1883..1916
                     /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    1923..1956
                     /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     2608..2627
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2867..2888
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2996..3029)
                     /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    3138..3726
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3191..3211)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3609..3620)
                     /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 cgacgccacc atggtggact catcacgtcg taagtggaat aagacaggtc
     1441 acgcagtcag agctataggt cggctgagct cactcgagaa cgtctatatc aaggccgaca
     1501 agcagaagaa cggcatcaag gcgaacttca agatccgcca caacatcgag gacggcggcg
     1561 tgcagctcgc ctaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1621 ccgacaacca ctacctgagc gtgcagtcca aactttcgaa agaccccaac gagaagcgcg
     1681 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1741 tgtacaaggg cggtaccgga gggagcatgg tgagcaaggt gagtacagga gggctagcat
     1801 aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat tatcactgtt
     1861 attctttaga taggtgcaaa gaataacttc gtataaggta tactatacga agttatggta
     1921 ccgaagttcc tattcttcaa atagtatagg aacttcaatt gttattggtg caaagaatca
     1981 cttccttaag tctttatact gcccaggaag ttcctattct tcaaaaggta taggaacttc
     2041 ggcgcgcctc acttcgctgt catcatttgt acaaactctt cgtagtttac ctgaccatcc
     2101 ccatcgatgt ctgcttccct gatcatttca tcaacctctt catctgttaa cttctctcca
     2161 aggtttgtca tcacgtggcg aagctctgct gcactgatgt agccattgcc atccttatca
     2221 aacacaccga acgcttctct aatttcttct tccgtgtccc tgtagctccc ttttcttgcc
     2281 atcattgtca ggaactcagg gaagtcgatt gtgccgtcac cgtcggcatc tacttcattg
     2341 atcatgtcct gcagctctgc ttctgtgggg ttctgcccca gagaccgcat caccgtcccc
     2401 agctccttgg ttgttattgt cccatccccg tccttgtcaa atagggagaa agcctcttta
     2461 aattctgcga tctgctcttc agtcagttgg tccggcaggt tgtactccag cttgtgcccc
     2521 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     2581 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     2641 gtgcgctcct ggatgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     2701 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     2761 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     2821 gcatcgccct caccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     2881 tcgaccagga tgggcaccac cccggtgaac agctcctcgc cctgtgggac gacatgactt
     2941 aaccagttaa ccatgttctc tgaacatgag aagggctggc cactctccat ctagagaagt
     3001 tcctatacta tttgaagaat aggaacttct tgccttaacc cagaaattat cactgttatt
     3061 ctttagaata gtgcaaagag aagttcctat accttttgaa gaataggaac ttcgaattcg
     3121 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3181 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3241 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3301 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3361 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3421 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3481 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3541 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     3601 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3661 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3721 ccccgcatcg ataccg
//
