LOCUS       sequence-265000-        3631 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of NpHR3.3-EYFP in cells expressing 
            Flp AND NOT Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3631)
  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 3631)
  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-NpHR3.3-EYFP
FEATURES             Location/Qualifiers
     source          1..3631
                     /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(703..954)
                     /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(745..766)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1094..1113
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1335..1368)
                     /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(1419..1452)
                     /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     1565..1586
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1629..1685)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    2490..2523
                     /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    2574..2607
                     /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(2891..2924)
                     /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    3033..3621
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3086..3106)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3504..3515)
                     /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 acacctcgtt ctcgtagcag aacttgtaca gctcgtccat
      721 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      781 gttggggtct ttgctcaggg cggactggta gctcaggtag tggttgtcgg gcagcagcac
      841 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
      901 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
      961 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     1021 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     1081 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     1141 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     1201 ggcgaagcac tgcaggccgt agccgaaggt ggtcacgagg gtgggccagg gcacgggcag
     1261 ctgtgggacg acatgactta accagttaac catgttctct gaacatgaga agggctggcc
     1321 actctccagt cgacataact tcgtatagga tactttatac gaagttatgc agaatggtag
     1381 ctggattgta gctgctatta gcaatatgaa acctcttaat aacttcgtat agtatacctt
     1441 atacgaagtt atactagtcc tcctgtactc accttgccgg tggtgcagat gaacttcagg
     1501 gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg
     1561 tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc
     1621 ttgctcacgg gaccggggtt ttcttccacg tctcctgctt gctttaacag agagaagttc
     1681 gtggctccgg agccgggcgc gcccacgttg atgtcgatct ggtccagggg gatgtactcg
     1741 ccctcgctgg tgatcctgct cttatcatca gccggggtcc cagaagcaga tggaacatcc
     1801 aaaatgctgc cggacacgac agattcgttt gacgtcagat agttcaggag caagaatgcg
     1861 aaaatgtact ttgccacgat atccaggaaa gaatatcccc agctcgtcac cccaacgggc
     1921 aacaccgcga taccctcgac tccaagagcc cacacaattg gataccccag ccacatcact
     1981 actgtcaaca gcttcagggt attgaacatg tcagcggttc ccgcggcttt ggcgtcctgt
     2041 gcccactcca ccagcaggat atacaagacc accagaaagc atgcacaact gatagcgtaa
     2101 aagaaccatc tcatgaggtg ggaggaggta gtcagggccg cggcaaggcc agtcacgcac
     2161 atagcgatat cgaaagtgat agctgtgaag agctttgtag cattgcttcc ggccaggaga
     2221 cccagagcga ggagaatcat gggcgtggaa agtgcccagg tgagataccg tccccacatg
     2281 gtgactacac catctacctc ttctcctccc agcatcactg agctgccttc tgcaaaatgg
     2341 cccgctggca tttcaagaac ggaaattgtc aggccgctcg ccaatcctaa acacatgaga
     2401 aatagagtca taagcacatg ggtcacatac aggagatatg tatataacat taatacaatt
     2461 ttataaaaaa aagggggggc acaggtacca taacttcgta taaagtatcc tatacgaagt
     2521 tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt
     2581 cgtataaggt atactatacg aagttatgct agcaaccccg acacttacct gtgtaggagg
     2641 caatgctgac gacaggcaca aggatggttg acacagcaat aagttttgcc cgtggatcat
     2701 cgagtcctcg ggtcataaaa acgaacagca gtatactcag tcctgcaagt gcgatgttga
     2761 tatagagact gcttgcaagc aaagggtcgt tcagcacgaa ctcgaacaac tccctttggg
     2821 taacctcggc ttgaagggcc acggcactct cggtcacggg aggcagggtc tctgtcatgg
     2881 tggcgagctc gaagttccta tactatttga agaataggaa cttcttgcct taacccagaa
     2941 attatcactg ttattcttta gaatggtgca aagagaagtt cctatacctt ttgaagaata
     3001 ggaacttcga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     3061 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3121 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3181 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3241 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3301 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3361 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3421 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     3481 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3541 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3601 ccctttgggc cgcctccccg catcgatacc g
//
