LOCUS Exported 3679 bp ds-DNA linear SYN 31-MAY-2021 DEFINITION Intersectional viral expression of ChRmine-oScarlet in cells expressing both Cre AND Flp. ACCESSION . VERSION . KEYWORDS pAAV-nEF-Con/Fon-ChRmine-oScarlet SOURCE synthetic DNA construct ORGANISM synthetic DNA construct 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 May 31, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3679 /organism="synthetic DNA construct" /mol_type="other DNA" 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" primer_bind complement(981..1000) /label=mCherry-F /note="mCherry, forward primer" protein_bind 1264..1297 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 1348..1381 /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." CDS 2164..2220 /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" primer_bind complement(2354..2372) /label=mCherry-R /note="mCherry, reverse primer" protein_bind complement(2453..2486) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(2537..2570) /label=loxN /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTATACCT). loxN sites are compatible with each other, but incompatible with loxP or lox2272 sites." regulatory 2923..2932 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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 tactagttgt gccccccctt ttttttataa aattgtatta atgttatata catatctcct 1441 gtatgtgacc catgtgctta tgactctatt tctcatgtgt ttaggtatgc cgactacatg 1501 ctgacctgcc caatgctggt gtacgatctg ctgtatcagc tgagggcccc ctatcgcgtg 1561 tcttgtagcg ccatcatctt tgccatcctg atgtctggcg tgctggccga gttctacgca 1621 gagggcgacc ctaggctgag gaatggcgcc tacgcctggt atggctttgg ctgtttctgg 1681 tttatcttcg cctactctat cgtgatgagc atcgtggcca agcagtatag ccggctggcc 1741 cagctggccc aggatacagg cgccgagcac agcctgcacg tgctgaagtt cgccgtgttt 1801 accttctcca tgctgtggat tctgtttccc ctggtgtggg ccatctgccc tagaggcttc 1861 ggctggatcg acgataactg gacagaggtg gcccactgcg tgtgcgacat cgtggccaag 1921 tcctgttacg gctttgccct ggcccggttc agaaagacct atgatgagga gctgtttcgg 1981 ctgctggagc agctgggaca cgacgaggat gagttccaga agctggagct ggatatgagg 2041 ctgagcagca atggcgagcg cctgcggaga ctgtctgcgg ccgccaagag caggatcacc 2101 agcgagggcg agtacatccc cctggaccag atcgacatca acgtgggcgc gcccggctcc 2161 ggagccacga acttctctct gttaaagcaa gcaggagacg tggaagaaaa ccccggtccc 2221 atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag 2281 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag 2341 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac 2401 atcctgtccc ctcagttcat gtacggctcc aggtgagtac aggaggggta 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 //