LOCUS Exported 7109 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Intersectional viral expression of ChRmine-oScarlet in cells expressing Cre AND NOT Flp. ACCESSION . VERSION . KEYWORDS pAAV-nEF-Con/Foff 2.0-ChRmine-oScarlet SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7109) 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 7109) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7109 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 188..399 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 412..680 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" protein_bind 708..741 /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)" regulatory 834..843 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 1196..1229 /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." protein_bind 1280..1313 /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)." primer_bind 1394..1412 /label=mCherry-R /note="mCherry, reverse primer" CDS complement(1546..1602) /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 complement(2385..2418) /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." protein_bind complement(2469..2502) /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)." primer_bind 2766..2785 /label=mCherry-F /note="mCherry, forward primer" protein_bind complement(3094..3127) /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 3236..3824 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3289..3309) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3707..3718) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3856..4332 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4055..4074) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4372..4512 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4372..4501 /label=AAV2 ITR rep_origin 4587..5042 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4674..4693) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4884..4905 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5059..5078) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5178..5200 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5238..5256) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5324..5428 /gene="bla" /label=AmpR promoter CDS 5429..6289 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5647..5666) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6460..7048 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6949..6968 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagga tctgcgatcg ctccggtgcc 181 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 241 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 301 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 361 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 421 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 481 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 541 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 601 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 661 gttctgcgcc gttacagatc caagctgtga ccggcgccta cggatccgaa gttcctattc 721 tctagaaagt ataggaactt cgcagagtag tagctggatt gtagctgcta ttagcaatct 781 aaaacctctt agaagttcct attcttcaaa aggtatagga acttcggcgc gccgccacca 841 tggcacacgc accaggcacc gaccagatgt tctacgtggg cacaatggac ggctggtatc 901 tggataccaa gctgaactcc gtggccatcg gcgcccactg gtcttgcttt atcgtgctga 961 caatcaccac attctacctg ggctatgagt cctggacatc tcggggccct agcaagagaa 1021 cctcctttta cgccggctat caggaggagc agaacctggc cctgtttgtg aatttctttg 1081 ccatgctgag ctacttcggc aagatcgtgg ccgacacact gggccacaac ttcggcgatg 1141 tgggcccctt catcatcggc ttcggcaatt acaggtaagt gtcggggttg agctcataac 1201 ttcgtatagt ataccttata cgaagttatt ctttgcacca ttctaaagaa taacagtgat 1261 aatttctggg ttaaggcaaa taacttcgta taggatactt tatacgaagt tatggtaccc 1321 ctcctgtact cacctggagc cgtacatgaa ctgaggggac aggatgtccc aggagaaggg 1381 cagggggcca cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc 1441 ctcgccctcg ccctcgatct cgaactcgtg gccgttcatg gagccctcca tgtgcacctt 1501 gaaccgcatg aactccttga tcactgcctc gcccttgctc accatgggac cggggttttc 1561 ttccacgtct cctgcttgct ttaacagaga gaagttcgtg gctccggagc cgggcgcgcc 1621 cacgttgatg tcgatctggt ccagggggat gtactcgccc tcgctggtga tcctgctctt 1681 ggcggccgca gacagtctcc gcaggcgctc gccattgctg ctcagcctca tatccagctc 1741 cagcttctgg aactcatcct cgtcgtgtcc cagctgctcc agcagccgaa acagctcctc 1801 atcataggtc tttctgaacc gggccagggc aaagccgtaa caggacttgg ccacgatgtc 1861 gcacacgcag tgggccacct ctgtccagtt atcgtcgatc cagccgaagc ctctagggca 1921 gatggcccac accaggggaa acagaatcca cagcatggag aaggtaaaca cggcgaactt 1981 cagcacgtgc aggctgtgct cggcgcctgt atcctgggcc agctgggcca gccggctata 2041 ctgcttggcc acgatgctca tcacgataga gtaggcgaag ataaaccaga aacagccaaa 2101 gccataccag gcgtaggcgc cattcctcag cctagggtcg ccctctgcgt agaactcggc 2161 cagcacgcca gacatcagga tggcaaagat gatggcgcta caagacacgc gatagggggc 2221 cctcagctga tacagcagat cgtacaccag cattgggcag gtcagcatgt agtcggcata 2281 cctaaacaca tgagaaatag agtcataagc acatgggtca catacaggag atatgtatat 2341 aacattaata caattttata aaaaaaaggg ggggcacaac tagtataact tcgtataagg 2401 tatactatac gaagttatta agaggtttca tattgctaat agcagctaca atccagctac 2461 cattctgcat aacttcgtat aaagtatcct atacgaagtt atgtcgactg gagagtggcc 2521 agcccttctc atgttcagag aacatggtta actggttaag tcatgtcgtc ccacagggcc 2581 ttcaccaagc accccgccga catccccgac tactataagc agtccttccc cgagggcttc 2641 aagtgggacc gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc 2701 tccctggagg acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct 2761 gacggccccg taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac 2821 cccgaggacg gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggg 2881 cgctacctgg cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc 2941 gcctacaacg tggaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg 3001 gaacagtacg aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag 3061 ttctgctacg agaacgaggt gtaatgagct agcgaagttc ctatactttc tagagaatag 3121 gaacttctaa gcagacaatc ttgtaataca tcagcagcta gtagattgct ccagctagaa 3181 gttcctatac cttttgaaga ataggaactt cgaattcgat atcaagctta tcgataatca 3241 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 3301 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 3361 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 3421 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 3481 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 3541 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 3601 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg 3661 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 3721 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 3781 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct 3841 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg 3901 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg 3961 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg 4021 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca 4081 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct 4141 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt 4201 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag 4261 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc 4321 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc 4381 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac 4441 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca 4501 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 4561 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg 4621 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 4681 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 4741 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 4801 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 4861 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 4921 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 4981 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat 5041 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca 5101 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag 5161 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa 5221 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 5281 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 5341 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 5401 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 5461 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 5521 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 5581 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 5641 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 5701 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 5761 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 5821 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 5881 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 5941 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 6001 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 6061 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 6121 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 6181 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 6241 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 6301 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 6361 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 6421 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 6481 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 6541 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 6601 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 6661 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 6721 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 6781 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 6841 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 6901 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 6961 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 7021 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 7081 ggccttttgc tggccttttg ctcacatgt //