LOCUS       sequence-264993-        3586 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of bREACHes-EYFP in cells expressing
            Flp AND NOT Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3586)
  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 3586)
  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-bREACHes-EYFP
FEATURES             Location/Qualifiers
     source          1..3586
                     /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     1544..1565
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    2236..2269
                     /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    2320..2353
                     /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      2830..2839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2846..2879)
                     /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    2988..3576
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3041..3061)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3459..3470)
                     /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 tactagtcct
     1441 cctgtactca ccttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
     1501 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
     1561 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccac gttgatgtcg
     1621 atctggtcca gggggatgta ctcgccctcg ctggtgatcc tgctcttggc ggccgcatct
     1681 tcttcttccg ccaccagggt ttccacttcc atttcctggc ccgcaatggt aattttctgt
     1741 tttttgcgaa tatcgccata cagcagaata tgttcatgaa ttttcacgcg cagatagttg
     1801 cccagcacgc cccacatgtt cttcgcaatc agatccagaa tgctatggcc aatcgcgctg
     1861 ccatacgggc taatatggcc gaagccctcg gggcccagca ggaacagcac ggggaacatg
     1921 ccccagctca cgaagaacag ccaggccatg gctctcacca gctgtctgca caggcctttc
     1981 ggcacggtat gaaacgcttc aatatacact ttggccgcat gaaaataggt atacatgcca
     2041 tagctcaggc taatcagaaa aaacagaatt ttggtccagc cggtgcacat cgcgctggtc
     2101 gcgccccaca caatgcagcc cacgtcgctc acctaaacac atgagaaata gagtcataag
     2161 cacatgggtc acatacagga gatatgtata taacattaat acaattttat aaaaaaaagg
     2221 gggggcacag gtaccataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc
     2281 cagaaattat cactgttatt ctttagaatg gtgcaaagaa taacttcgta taaggtatac
     2341 tatacgaagt tatgctagca accccgacac ttaccagcag gcccatggtg cgtttgctat
     2401 aatcatcttt caggccggtc aggttgctca gatgaatcag aatcacgggg caggtcagca
     2461 gccatgagcc atagcgcatc cacaccacgc cgttgccgct gctcagccac agggtcgccg
     2521 ggctatcaaa ttcatgaaac gcttcaataa tgcttttcat catttcaatc agcgccacat
     2581 acacttcttc ccagccgcag gtcgcgcgcc acgcctgata cgcataccag cccaggcacg
     2641 ccacgctcag cgcaaacacc acccactgca ggatgttggc ggccagcttc tcggcgttgg
     2701 tgccgttact cttcagccag gccaggcaga agcactggcc gttgttgggg atgcagatca
     2761 cgctgccgtt gttctccagg gtgtagctgg tctggaacag tccgacggca gacaaagcgc
     2821 cgccatagtc catggtggcg agctcgaagt tcctatacta tttgaagaat aggaacttct
     2881 tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagag aagttcctat
     2941 accttttgaa gaataggaac ttcgaattcg atatcaagct tatcgataat caacctctgg
     3001 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3061 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3121 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3181 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3241 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3301 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3361 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     3421 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3481 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3541 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccg
//
