LOCUS       sequence-264996-        3531 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of Arch3.3-EYFP in cells expressing 
            Flp AND NOT Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3531)
  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 3531)
  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-Arch3.3-p2a-EYFP
FEATURES             Location/Qualifiers
     source          1..3531
                     /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    2365..2398
                     /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    2449..2482
                     /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      2775..2784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2791..2824)
                     /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    2933..3521
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2986..3006)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3404..3415)
                     /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 cttgtcggcg gcactgacat cggcaccggc actgggttct
     1801 ggtgcctcgg tgtcgcccag aatagcccgg gatctcaaca ggataaagcc aaagccgacc
     1861 ttggcagtca cgtccaacac cataaacagc agagtttcga tgcccaggcc caccacgcca
     1921 gcgccctcag tgcctatgat ccacaggata gggtaagcgg tccacagcac caagaccaga
     1981 gctgtcaggg tgttaaaggt agatgccacc tcggggcccc gctcctttgc agcagatcgc
     2041 agggatgtag ccagaaaata gagcaccact atcatgcaaa ttgtagagaa caaccaccaa
     2101 ctgtatctgg ctatggccgt gtggctcaag gctccgatga ggccagtgac gatcatcagg
     2161 gcgtccacac ccaccagggt gccgatggtc acccgatcca ccttagcgag aagggccaga
     2221 tccagcagca gaagtggggt ggtaaacagc cagtcggcgt acctaaacac atgagaaata
     2281 gagtcataag cacatgggtc acatacagga gattgtatat aacattaata caattttata
     2341 aaaaaaaggg ggggcacagg taccataact tcgtataaag tatcctatac gaagttattt
     2401 gccttaaccc agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat
     2461 aaggtatact atacgaagtt atgctagcaa ccccgacact tacctggcat aatagatatc
     2521 caacatttcg cccccgacgg tcacctcagt aagcccgata ccaaagaaca tagacagata
     2581 tgcggcggat gcgattccgg gcaccaggat agtcacagcg taatattccc gggcatcctt
     2641 atcggtgact ccccatccgc ggaccagaaa gtagaaggtt ccaatcagca tcagcagagt
     2701 gccgatgccc agccacagag tttcaggtct gccgtcaccc agcaggtcgt aaccagcctg
     2761 cagagcgatg gggtccatgg tggcgagctc gaagttccta tactatttga agaataggaa
     2821 cttcttgcct taacccagaa attatcactg ttattcttta gaatggtgca aagagaagtt
     2881 cctatacctt ttgaagaata ggaacttcga attcgatatc aagcttatcg ataatcaacc
     2941 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3001 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3061 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3121 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3181 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3241 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3301 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3361 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3421 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3481 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc g
//
