LOCUS       sequence-265006-        3679 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Intersectional viral expression of ChRmine-p2a-oScarlet in cells 
            expressing Flp AND NOT Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3679)
  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 3679)
  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-ChRmine-oScarlet
FEATURES             Location/Qualifiers
     source          1..3679
                     /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)"
     primer_bind     complement(981..1000)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1264..1297)
                     /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(1348..1381)
                     /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     1462..1480
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1614..1670)
                     /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    2453..2486
                     /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    2537..2570
                     /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      2923..2932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2939..2972)
                     /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    3081..3669
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3134..3154)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3552..3563)
                     /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 ggcgcgcctc attacacctc gttctcgtag cagaacttgt acagctcgtc
      721 catgccgccg gtggagtggc ggccctcgga gcgttcgtac tgttccacca cggtgtagtc
      781 ctcgttgtgg gaggtgatgt ccaacttgcg gtccacgttg taggcgccgg gcatctgcac
      841 gggcttcttg gccttgtagg tggtcttgaa gtccgccagg tagcgcccgc cgtccttcag
      901 gcgcagggcc atcttaatgt cgcccttcag cacgccgtcc tcggggtaca accgctcggt
      961 ggacgcttcc cagcccattg tcttcttctg cattacgggg ccgtcaggag ggaagttggt
     1021 gccgcggagc ttcaccttgt agatcagggt gccgtcctcc agggaggtgt cctgggtcac
     1081 ggtcacggcg ccgccgtcct cgaagttcat cacgcggtcc cacttgaagc cctcggggaa
     1141 ggactgctta tagtagtcgg ggatgtcggc ggggtgcttg gtgaaggccc tgtgggacga
     1201 catgacttaa ccagttaacc atgttctctg aacatgagaa gggctggcca ctctccagtc
     1261 gacataactt cgtataggat actttatacg aagttatgca gaatggtagc tggattgtag
     1321 ctgctattag caatatgaaa cctcttaata acttcgtata gtatacctta tacgaagtta
     1381 tactagtcct cctgtactca cctggagccg tacatgaact gaggggacag gatgtcccag
     1441 gagaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag
     1501 gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga gccctccatg
     1561 tgcaccttga accgcatgaa ctccttgatc actgcctcgc ccttgctcac catgggaccg
     1621 gggttttctt ccacgtctcc tgcttgcttt aacagagaga agttcgtggc tccggagccg
     1681 ggcgcgccca cgttgatgtc gatctggtcc agggggatgt actcgccctc gctggtgatc
     1741 ctgctcttgg cggccgcaga cagtctccgc aggcgctcgc cattgctgct cagcctcata
     1801 tccagctcca gcttctggaa ctcatcctcg tcgtgtccca gctgctccag cagccgaaac
     1861 agctcctcat cataggtctt tctgaaccgg gccagggcaa agccgtaaca ggacttggcc
     1921 acgatgtcgc acacgcagtg ggccacctct gtccagttat cgtcgatcca gccgaagcct
     1981 ctagggcaga tggcccacac caggggaaac agaatccaca gcatggagaa ggtaaacacg
     2041 gcgaacttca gcacgtgcag gctgtgctcg gcgcctgtat cctgggccag ctgggccagc
     2101 cggctatact gcttggccac gatgctcatc acgatagagt aggcgaagat aaaccagaaa
     2161 cagccaaagc cataccaggc gtaggcgcca ttcctcagcc tagggtcgcc ctctgcgtag
     2221 aactcggcca gcacgccaga catcaggatg gcaaagatga tggcgctaca agacacgcga
     2281 tagggggccc tcagctgata cagcagatcg tacaccagca ttgggcaggt cagcatgtag
     2341 tcggcatacc taaacacatg agaaatagag tcataagcac atgggtcaca tacaggagat
     2401 atgtatataa cattaataca attttataaa aaaaaggggg ggcacaggta ccataacttc
     2461 gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt
     2521 tagaatggtg caaagaataa cttcgtataa ggtatactat acgaagttat gctagcaacc
     2581 ccgacactta cctgtaattg ccgaagccga tgatgaaggg gcccacatcg ccgaagttgt
     2641 ggcccagtgt gtcggccacg atcttgccga agtagctcag catggcaaag aaattcacaa
     2701 acagggccag gttctgctcc tcctgatagc cggcgtaaaa ggaggttctc ttgctagggc
     2761 cccgagatgt ccaggactca tagcccaggt agaatgtggt gattgtcagc acgataaagc
     2821 aagaccagtg ggcgccgatg gccacggagt tcagcttggt atccagatac cagccgtcca
     2881 ttgtgcccac gtagaacatc tggtcggtgc ctggtgcgtg tgccatggtg gcgagctcga
     2941 agttcctata ctatttgaag aataggaact tcttgcctta acccagaaat tatcactgtt
     3001 attctttaga atggtgcaaa gagaagttcc tatacctttt gaagaatagg aacttcgaat
     3061 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     3121 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3181 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3241 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3301 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3361 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3421 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3481 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     3541 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3601 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3661 cctccccgca tcgataccg
//
