LOCUS       sequence-265003-        3598 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of iC++-EYFP in cells expressing Flp
            AND NOT Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3598)
  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 3598)
  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-iC++-EYFP
FEATURES             Location/Qualifiers
     source          1..3598
                     /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    2197..2230
                     /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    2281..2314
                     /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      2842..2851
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2858..2891)
                     /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    3000..3588
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3053..3073)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3471..3482)
                     /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 ggccgcgacc
     1681 gcgccagcct cggcctcgtc ctccaccagc gtctcgacct caatctcagt gccaccaatg
     1741 ttcaatttgg tggtcttgcg aatgtcgccg tggatgagga tatggctgtg gatcagcacg
     1801 cgcaggtagt ggccgagcag accccagcac tgcttcgaca tcaggtcaat gatggtgtgg
     1861 ccgacgttgg agccgtacct gctcaggacg ccgaagccct cggggccgag gatgaacagg
     1921 atggggaaca taccccatga tacgaagaag agccaagcca tgccagtcac cacctggcga
     1981 caccggccct tcggcacggt gtggtacgcc tcaatgtaga ccttggctgc gttgaagaat
     2041 gtgtagatgc cgtagcacag gcccatcagg aagaaaatga cacggacgta tccctaaaca
     2101 catgagaaat agagtcataa gcacatgggt cacatacagg agatatgtat ataacattaa
     2161 tacaatttta taaaaaaaag ggggggcaca ggtaccataa cttcgtataa agtatcctat
     2221 acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga
     2281 ataacttcgt ataaggtata ctatacgaag ttatgctagc aaccccgaca cttaccttgg
     2341 acagcgcggc cgtggtgccc cacacgatcg tgccgatatc tgacaccagc agacccatgg
     2401 tacgcttgtt atagtcgttc gccagacccg taaggttgct caggtggata agaatgacag
     2461 ggcaggtcag cagccagctc gcgtaacgaa gccacctggt cttgttgccg ttggatgagt
     2521 agatcaccgc aggttcgtca aagctatgga aatactcgat gatgaacttg atcatctgga
     2581 tcgtggccac gtaaatattc tcccagccgc aagtagactt ccaggtctgg tagccgtaga
     2641 acatcaggca gagcgctgaa agcgcaaaag aaatccactg cagaatgttg gcagccaact
     2701 tctcggcatt tgttccgttg gatttaagcc aagccaagca gaagcactgg ccgttgttcg
     2761 ggatgcaaat aacagaacca ttgttctcaa gagtgtatga ggtttggaag agtccgacgg
     2821 cagacaaagc gccgccatag tccatggtgg cgagctcgaa gttcctatac tatttgaaga
     2881 ataggaactt cttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     2941 agaagttcct ataccttttg aagaatagga acttcgaatt cgatatcaag cttatcgata
     3001 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3061 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3121 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3181 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3241 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3301 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3361 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3421 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3481 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3541 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccg
//
