LOCUS       sequence-264972-        3737 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of sRGECO in cells expressing both 
            Cre AND Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3737)
  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 3737)
  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-sRGECO
FEATURES             Location/Qualifiers
     source          1..3737
                     /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."
     CDS             complement(1683..1739)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     regulatory      1751..1760
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1765..1798
                     /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    1849..1882
                     /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    1889..1922
                     /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     complement(2482..2501)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2997..3030)
                     /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    3139..3727
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3192..3212)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3610..3621)
                     /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 gtacctccct tgtacagctc gtccatgccg
     1441 ccggtggagt ggcggccctc ggcgcgttcg cactgttcca cgatggtgta gtcctcgttg
     1501 tgggacacga tgtccaactt gatgtccacg atgtaggcgc cgggcagctg cacgggcttc
     1561 ttggccttgt aggtggtctt gacctcggcg gcgtagtggc cgccgtcctt cagcctcagc
     1621 cccttcttga tctcgctctt cagggcgccg tcctcggggt acatccgctc ggaaaccacg
     1681 ggtgagctca gccgacctat agctctgact gcgtgacctg ccttattcca cttacgacgt
     1741 gatgagtcca ccatggtggc tagcataact tcgtataaag tatcctatac gaagttattt
     1801 gccttaaccc agaaattatc actgttattc tttagatagg tgcaaagaat aacttcgtat
     1861 aaggtatact atacgaagtt atggtaccga agttcctatt cttcaaatag tataggaact
     1921 tcaattgtta ttggtgcaaa gaatcacttc cttaagtctt tatactgccc aggaagttcc
     1981 tattcttcaa aaggtatagg aacttcggcg cgccctactt cgctgtcatc atttgtacaa
     2041 actcttcgta gtttacctga ccatccccat cgatgtctgc taccctgatc atttcatcaa
     2101 cctcctcatc tgttaacttc tctccaaggt ctgtcatcac gtggcgaagc tctgctgcgc
     2161 cgatgtagcc attgccgtcc ttatcaaaca cgcggaacgc ttcgcgaatt tcctcttcac
     2221 tgtctgtgtc attcattttt cttgccatca tcgtcaggaa ctcagggaag tcgaatgtgc
     2281 cgtcaccgtc ggcatctact tcattgatca tgtcctgcag ctctgcttct gtggggttct
     2341 gccccagcga tcggaacacc gtcccaagct ccttggttgt tatcgtccca tccccgtcct
     2401 tgtcaaatag ggagaaagcc tctttaaatt ctgcgatctg ctcttcagtc aggtcgtcac
     2461 gcgtagcctc ccagcccatg gtcttcttct gcattacggg gccgtcgggg gggaagttgg
     2521 tgccgcgcag cttcaccttg tagatgaata cgccgtcctg cagggaggag tcctggttca
     2581 cgtgaataat gccgccgtcc tcgaagttca tcacgcggtc ccacctgaag ccctcgggga
     2641 aggacagctt gaagtagtcg gggatgtcgg ctgggtgctt aatgtaggcc ttggagccgt
     2701 acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc ttggtcacct
     2761 tcagcttagc ggtctgaaag gcctcgtagg ggcggccctc gccctcgccc tcgatctcga
     2821 actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac tccttgatga
     2881 tggccatgtt atcctcctcg cccttgctca ccagactccc gcctgtggga cgacatgact
     2941 taaccagtta accatgttct ctgaacatga gaagggctgg ccactctcca tctagagaag
     3001 ttcctatact atttgaagaa taggaacttc ttgccttaac ccagaaatta tcactgttat
     3061 tctttagaat agtgcaaaga gaagttccta taccttttga agaataggaa cttcgaattc
     3121 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3181 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3241 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3301 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3361 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3421 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3481 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3541 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     3601 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3661 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3721 tccccgcatc gataccg
//
