LOCUS       sequence-264991-        3585 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of bREACHes-EYFP in cells expressing
            both Cre AND Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3585)
  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 3585)
  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/Fon-bREACHes-EYFP
FEATURES             Location/Qualifiers
     source          1..3585
                     /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."
     primer_bind     complement(2101..2122)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    2235..2268
                     /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    2319..2352
                     /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      2829..2838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2845..2878)
                     /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    2987..3575
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3040..3060)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3458..3469)
                     /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 tactagttgt
     1441 gccccccctt ttttttataa attgtattaa tgttatatac atatctcctg tatgtgaccc
     1501 atgtgcttat gactctattt ctcatgtgtt taggtgagcg acgtgggctg cattgtgtgg
     1561 ggcgcgacca gcgcgatgtg caccggctgg accaaaattc tgttttttct gattagcctg
     1621 agctatggca tgtataccta ttttcatgcg gccaaagtgt atattgaagc gtttcatacc
     1681 gtgccgaaag gcctgtgcag acagctggtg agagccatgg cctggctgtt cttcgtgagc
     1741 tggggcatgt tccccgtgct gttcctgctg ggccccgagg gcttcggcca tattagcccg
     1801 tatggcagcg cgattggcca tagcattctg gatctgattg cgaagaacat gtggggcgtg
     1861 ctgggcaact atctgcgcgt gaaaattcat gaacatattc tgctgtatgg cgatattcgc
     1921 aaaaaacaga aaattaccat tgcgggccag gaaatggaag tggaaaccct ggtggcggaa
     1981 gaagaagatg cggccgccaa gagcaggatc accagcgagg gcgagtacat ccccctggac
     2041 cagatcgaca tcaacgtggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2101 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2161 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caaggtgagt
     2221 acaggagggg taccataact tcgtataaag tatcctatac gaagttattt gccttaaccc
     2281 agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat aaggtatact
     2341 atacgaagtt atgctagcaa ccccgacact taccagcagg cccatggtgc gtttgctata
     2401 atcatctttc aggccggtca ggttgctcag atgaatcaga atcacggggc aggtcagcag
     2461 ccatgagcca tagcgcatcc acaccacgcc gttgccgctg ctcagccaca gggtcgccgg
     2521 gctatcaaat tcatgaaacg cttcaataat gcttttcatc atttcaatca gcgccacata
     2581 cacttcttcc cagccgcagg tcgcgcgcca cgcctgatac gcataccagc ccaggcacgc
     2641 cacgctcagc gcaaacacca cccactgcag gatgttggcg gccagcttct cggcgttggt
     2701 gccgttactc ttcagccagg ccaggcagaa gcactggccg ttgttgggga tgcagatcac
     2761 gctgccgttg ttctccaggg tgtagctggt ctggaacagt ccgacggcag acaaagcgcc
     2821 gccatagtcc atggtggcga gctcgaagtt cctatactat ttgaagaata ggaacttctt
     2881 gccttaaccc agaaattatc actgttattc tttagaatgg tgcaaagaga agttcctata
     2941 ccttttgaag aataggaact tcgaattcga tatcaagctt atcgataatc aacctctgga
     3001 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3061 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3121 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3181 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3241 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3301 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3361 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3421 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3481 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3541 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccg
//
