LOCUS       sequence-264999-        3634 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of NpHR3.3-EYFP in cells expressing 
            Cre AND NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3634)
  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 3634)
  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-NpHR3.3-EYFP
FEATURES             Location/Qualifiers
     source          1..3634
                     /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)"
     protein_bind    complement(956..989)
                     /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(1040..1073)
                     /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     1186..1207
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1250..1306)
                     /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    2111..2144
                     /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    2195..2228
                     /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(2450..2469)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2609..2860
                     /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     2797..2818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2894..2927)
                     /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    3036..3624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3089..3109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3507..3518)
                     /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 caccatgaca gagaccctgc ctcccgtgac cgagagtgcc
      721 gtggcccttc aagccgaggt tacccaaagg gagttgttcg agttcgtgct gaacgaccct
      781 ttgcttgcaa gcagtctcta tatcaacatc gcacttgcag gactgagtat actgctgttc
      841 gtttttatga cccgaggact cgatgatcca cgggcaaaac ttattgctgt gtcaaccatc
      901 cttgtgcctg tcgtcagcat tgcctcctac acaggtaagt gtcggggttg agctcataac
      961 ttcgtatagt ataccttata cgaagttatt ctttgcacca ttctaaagaa taacagtgat
     1021 aatttctggg ttaaggcaaa taacttcgta taggatactt tatacgaagt tatggtaccc
     1081 ctcctgtact caccttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     1141 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     1201 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacg ggaccggggt
     1261 tttcttccac gtctcctgct tgctttaaca gagagaagtt cgtggctccg gagccgggcg
     1321 cgcccacgtt gatgtcgatc tggtccaggg ggatgtactc gccctcgctg gtgatcctgc
     1381 tcttatcatc agccggggtc ccagaagcag atggaacatc caaaatgctg ccggacacga
     1441 cagattcgtt tgacgtcaga tagttcagga gcaagaatgc gaaaatgtac tttgccacga
     1501 tatccaggaa agaatatccc cagctcgtca ccccaacggg caacaccgcg ataccctcga
     1561 ctccaagagc ccacacaatt ggatacccca gccacatcac tactgtcaac agcttcaggg
     1621 tattgaacat gtcagcggtt cccgcggctt tggcgtcctg tgcccactcc accagcagga
     1681 tatacaagac caccagaaag catgcacaac tgatagcgta aaagaaccat ctcatgaggt
     1741 gggaggaggt agtcagggcc gcggcaaggc cagtcacgca catagcgata tcgaaagtga
     1801 tagctgtgaa gagctttgta gcattgcttc cggccaggag acccagagcg aggagaatca
     1861 tgggcgtgga aagtgcccag gtgagatacc gtccccacat ggtgactaca ccatctacct
     1921 cttctcctcc cagcatcact gagctgcctt ctgcaaaatg gcccgctggc atttcaagaa
     1981 cggaaattgt caggccgctc gccaatccta aacacatgag aaatagagtc ataagcacat
     2041 gggtcacata caggagatat gtatataaca ttaatacaat tttataaaaa aaaggggggg
     2101 cacaactagt ataacttcgt ataaggtata ctatacgaag ttattaagag gtttcatatt
     2161 ggtaatagca gctacaatcc agctaccatt ctgcataact tcgtataaag tatcctatac
     2221 gaagttatgt cgactggaga gtggccagcc cttctcatgt tcagagaaca tggttaactg
     2281 gttaagtcat gtcgtcccac agctgcccgt gccctggccc accctcgtga ccaccttcgg
     2341 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2401 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2461 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2521 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2581 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2641 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2701 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     2761 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2821 cgccgccggg atcactctcg gcatggacga gctgtacaag ttctgctacg agaacgaggt
     2881 gtaatgagct agcgaagttc ctatactttc tagagaatag gaacttctaa gcagacaatc
     2941 ttgtaataca tcagcagcta gtagattgct ccagctagaa gttcctatac cttttgaaga
     3001 ataggaactt cgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     3061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3421 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3601 tctccctttg ggccgcctcc ccgcatcgat accg
//
