LOCUS       sequence-264987-        3547 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of ChR2(ET/TC)-EYFP in cells 
            expressing Flp AND NOT Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3547)
  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 3547)
  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-nEF-Coff/Fon-ChR2(ET/TC)-EYFP
FEATURES             Location/Qualifiers
     source          1..3547
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        33..244
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             257..525
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    553..586
                     /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)"
     CDS             complement(682..933)
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(724..745)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1073..1092
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1314..1347)
                     /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(1398..1431)
                     /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      1607..1616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2238..2271
                     /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    2322..2355
                     /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      2791..2800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2807..2840)
                     /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    2949..3537
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3002..3022)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3420..3431)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
       61 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
      121 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
      181 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
      241 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
      301 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
      361 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
      421 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
      481 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
      541 gcctacggat ccgaagttcc tattcttcaa atagtatagg aacttcgcag agtagtagct
      601 ggattgtagc tgctattagc aatctaaaac ctcttagaag ttcctattct tcaaaaggta
      661 taggaacttc ggcgcgcctt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
      721 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
      781 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
      841 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
      901 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
      961 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     1021 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     1081 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     1141 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta
     1201 gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc tgtgggacga catgacttaa
     1261 ccagttaacc atgttctctg aacatgagaa gggctggcca ctctccagtc gacataactt
     1321 cgtataggat actttatacg aagttatgca gaatggtagc tggattgtag ctgctattag
     1381 caatatgaaa cctcttaata acttcgtata gtatacctta tacgaagtta tactagtcct
     1441 cctgtactca ccttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
     1501 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcacc gtccagctcg
     1561 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcggcc
     1621 gctggcacgg ctccggcctc ggcttcgtct tcgacgagag tctcgacctc gatctccgtt
     1681 ccgccgatgt tcagtttggt ggttttgcgg atatctccgt gaatcaatat gtgctcgtgg
     1741 atcaggacgc gcaggtagtg tcccaacaac ccccaacaat ttttactcat cagatcaata
     1801 atcgtgtgac ctacggtgga gccatagacg ctcaggacgc caaaaccttc gggccccaaa
     1861 atgaagagaa ttgggaacat accccagctc acgaaaaaca gccatgccat gccggtcacg
     1921 acctggcggc accgaccctt tggcacagta tgataaccct cgatatatgc tttggcggcg
     1981 tgaaaaaatg tgttcgcgcc atagcacaat ccaagacaaa agaagatgac tttaacatag
     2041 ccggttgcca tggcgctggt agccccccac acgatacacc cgatgtctga gacaaggagt
     2101 cccatggttc tcctgctgta gtcgttgctc aggcctaaac acatgagaaa tagagtcata
     2161 agcacatggg tcacatacag gagatatgta tataacatta atacaatttt ataaaaaaaa
     2221 gggggggcac aggtaccata acttcgtata aagtatccta tacgaagtta tttgccttaa
     2281 cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg tataaggtat
     2341 actatacgaa gttatgctag caaccccgac acttaccggt gaggttgctc aggtggataa
     2401 ggatgacagg acaagtgagc agccaggttg catagcgcag ccactgcacc cggtgtcctg
     2461 tggcaaggta gagcatagag ggattcttaa actcaaaaaa gaactcgaga atcaccttaa
     2521 ccatttcaat ggcgcacaca tagatctcct cccagccgca tgtagatttc caggtttggt
     2581 aggcatagaa catcagcagc aaaatgctga atcctgctgc aagccactgc aggacatttg
     2641 acgcggtctg agcgccgttc gtgccgcgag attcaatcca tccggcacag taacattgat
     2701 cctcagggac caggacggac ccgttcacca ccacaggatt agtaacgaac aaaagttcgc
     2761 gtccgacggc agacaaagcg ccgccatagt ccatggtggc gagctcgaag ttcctatact
     2821 atttgaagaa taggaacttc ttgccttaac ccagaaatta tcactgttat tctttagaat
     2881 ggtgcaaaga gaagttccta taccttttga agaataggaa cttcgaattc gatatcaagc
     2941 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3001 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3061 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3121 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3181 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3241 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3301 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3361 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3421 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3481 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3541 gataccg
//
