LOCUS       sequence-265008-        3195 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of EYFP in cells expressing Cre AND 
            NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3195)
  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 3195)
  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-Ef1a-Con/Foff 2.0-EYFP
FEATURES             Location/Qualifiers
     source          1..3195
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        87..1264
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1255
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1212..1232
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1271..1304)
                     /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(1355..1388)
                     /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     1501..1522
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1564..1573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1580..1613
                     /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    1664..1697
                     /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    1704..1737
                     /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)"
     primer_bind     complement(2045..2064)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2204..2455
                     /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     2392..2413
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2465..2498)
                     /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    2607..3195
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2660..2680)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3078..3089)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccactggctg cagtacgtga ttcttgatcc
      421 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      481 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      541 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      601 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      661 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      721 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      781 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
      841 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
      901 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
      961 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1021 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1081 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1141 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1201 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1261 gtgaggatcc ataacttcgt ataggatact ttatacgaag ttatgtagct ggagcaattt
     1321 gaaaagattg ttgtattacc tctagctgcg cttaataact tcgtatagta taccttatac
     1381 gaagttatgt cgaccctcct gtactcacct tgccggtggt gcagatgaac ttcagggtca
     1441 gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta
     1501 cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc
     1561 tcaccatggt ggcactagta taacttcgta taaagtatcc tatacgaagt tatttgcctt
     1621 aacccagaaa ttatcactgt tattctttag ataggtgcaa agaataactt cgtataaggt
     1681 atactatacg aagttatggt accgaagttc ctattctcta gaaagtatag gaacttcgca
     1741 gaatggtagc tggattgtag ctgctattag caatatgaaa cctcttagaa gttcctattc
     1801 ttcaaaaggt ataggaactt cggcgcgcct ggagagtggc cagcccttct catgttcaga
     1861 gaacatggtt aactggttaa gtcatgtcgt cccacagctg cccgtgccct ggcccaccct
     1921 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1981 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2041 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2101 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2161 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2221 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2281 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2341 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2401 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagc
     2461 tagcgaagtt cctatacttt ctagagaata ggaacttcta agcagacaat cttgtaatac
     2521 atcagcagct agtagattgc tccagctaga agttcctata ccttttgaag aataggaact
     2581 tcgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2641 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2701 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2761 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2821 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2881 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2941 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3001 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     3061 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3121 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3181 gggccgcctc cccgc
//
