LOCUS       sequence-265009-        3540 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of ChR2-EYFP in cells expressing Cre
            AND NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3540)
  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 3540)
  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-Con/Foff 2.0-ChR2-EYFP
FEATURES             Location/Qualifiers
     source          1..3540
                     /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=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1124..1157)
                     /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    complement(1208..1241)
                     /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)."
     primer_bind     1354..1375
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    2041..2074
                     /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    2125..2158
                     /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)."
     primer_bind     complement(2380..2399)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2539..2790
                     /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     2727..2748
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2800..2833)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    2942..3530
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2995..3015)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3413..3424)
                     /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 tattctctag aaagtatagg aacttcgcag agtagtagct
      601 ggattgtagc tgctattagc aatctaaaac ctcttagaag ttcctattct tcaaaaggta
      661 taggaacttc ggcgcgccgc caccatggac tatggcggcg ctttgtctgc cgtcggacgc
      721 gaacttttgt tcgttactaa tcctgtggtg gtgaacgggt ccgtcctggt ccctgaggat
      781 caatgttact gtgccggatg gattgaatct cgcggcacga acggcgctca gaccgcgtca
      841 aatgtcctgc agtggcttgc agcaggattc agcattttgc tgctgatgtt ctatgcctac
      901 caaacctgga aatctacatg cggctgggag gagatctatg tgtgcgccat tgaaatggtt
      961 aaggtgattc tcgagttctt ttttgagttt aagaatccct ctatgctcta ccttgccaca
     1021 ggacaccggg tgcagtggct gcgctatgca gagtggctgc tcacttgtcc tgtcatcctt
     1081 atccgcctga gcaacctcac cggtaagtgt cggggttgag ctcataactt cgtatagtat
     1141 accttatacg aagttattct ttgcaccatt ctaaagaata acagtgataa tttctgggtt
     1201 aaggcaaata acttcgtata ggatacttta tacgaagtta tggtacccct cctgtactca
     1261 ccttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     1321 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     1381 gcaccacccc ggtgaacagc tcctcgccct tgctcacggc ggccgctggc acggctccgg
     1441 cctcggcttc gtcttcgacg agagtctcga cctcgatctc cgttccgccg atgttcagtt
     1501 tggtggtttt gcggatatct ccgtgaatca atatgtgctc gtggatcagg acgcgcaggt
     1561 agtgtcccaa caacccccaa caatttttac tcatcagatc aataatcgtg tgacctacgg
     1621 tggagccata gacgctcagg acgccaaaac cttcgggccc caaaatgaag agaattggga
     1681 acatacccca gctcacgaaa aacagccatg ccatgccggt cacgacctgg cggcaccgac
     1741 cctttggcac agtatgataa ccctcgatat atgctttggc ggcgtgaaaa aatgtgttcg
     1801 cgccatagca caatccaaga caaaagaaga tgactttaac atagccggtt gccatggcgc
     1861 tggtagcccc ccacacgata gtcccgatgt ctgagacaag gagtcccatg gttctcctgc
     1921 tgtagtcgtt gctcaggcct aaacacatga gaaatagagt cataagcaca tgggtcacat
     1981 acaggagata tgtatataac attaatacaa tttataaaaa aaaggggggg cacaactagt
     2041 ataacttcgt ataaggtata ctatacgaag ttattaagag gtttcatatt gctaatagca
     2101 gctacaatcc agctaccatt ctgcataact tcgtataaag tatcctatac gaagttatgt
     2161 cgactggaga gtggccagcc cttctcatgt tcagagaaca tggttaactg gttaagtcat
     2221 gtcgtcccac agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     2281 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2341 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2401 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2461 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2521 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2581 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2641 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     2701 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2761 atcactctcg gcatggacga gctgtacaag taagctagcg aagttcctat actttctaga
     2821 gaataggaac ttctaagcag acaatcttgt aatacatcag cagctagtag attgctccag
     2881 ctagaagttc ctataccttt tgaagaatag gaacttcgaa ttcgatatca agcttatcga
     2941 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3001 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     3061 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     3121 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3181 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3241 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3301 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3361 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3421 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3481 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
//
